Information on EC 3.4.21.62 - Subtilisin

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY
3.4.21.62
-
RECOMMENDED NAME
GeneOntology No.
Subtilisin
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyses peptide amides
show the reaction diagram
hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyzes peptide amides. Mechanism
-
Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyses peptide amides
show the reaction diagram
importance of tetrahedral intermediate formation in the catalytic mechanism of the serine protease subtilisin. Substrate binding induces the formation of a strong hydrogen bond or low-barrier hydrogen bond between histidine-57 and aspartate-102 that increases the pK(a) of the active site histidine, allowing it to be an effective general base catalyst for the formation of the tetrahedral intermediate and increasing the effective molarity of the catalytic hydroxyl group of serine-195, catalytic mechanism, overview
-
Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyses peptide amides
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ester bond hydrolysis
-
-
-
-
ester bond hydrolysis
-
-
ester bond hydrolysis
-
-
hydrolysis of peptide bond
-
-
-
-
transesterification
-
-
-
-
transpeptidation
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
AcpII
Alkalimonas collagenimarina AC40T
-
-
Ak.1 protease
-
-
Ak.1 protease
Bacillus sp. Ak.1
-
-
-
Alcalase
-
-
-
-
Alcalase
synthetic construct
-
-
Alcalase 0.6L
-
-
-
-
Alcalase 2.5L
-
-
-
-
ALE1 subtilase
-
-
ALK-enzyme
-
-
-
-
Alkaline mesentericopeptidase
-
-
-
-
Alkaline protease
-
-
-
-
Alkaline protease
Alkalimonas collagenimarina AC40T
-
-
Alzwiprase
-
-
Asp v 13
-
subtilisin-like serine protease
Bacillopeptidase A
-
-
-
-
Bacillopeptidase B
-
-
-
-
Bacillus gibsonii alkaline protease
GN111900.1
-
Bacillus gibsonii alkaline protease
Bacillus gibsonii DSM 14391
GN111900.1
-
-
Bacillus subtilis alkaline proteinase Bioprase
-
-
-
-
BgAP
GN111900.1
-
BgAP
Bacillus gibsonii DSM 14391
GN111900.1
-
-
Bioprase AL 15
-
-
-
-
Bioprase APL 30
-
-
-
-
C1 subtilase
-
-
Colistinase
-
-
-
-
EC 3.4.21.14
-
-
formerly
-
EC 3.4.21.14
-
formerly included in
EC 3.4.21.14
-
formerly
EC 3.4.21.14
-
formerly included in
EC 3.4.4.16
-
-
formerly
-
Esperase
-
-
-
-
Fe protease
-
-
Fe protease
Fusarium equiseti CBS119568
-
-
-
Genenase I
-
-
-
-
intracellular subtilisin protease
-
Kazusase
-
-
-
-
Maxatase
-
-
-
-
Nagarse
-
-
-
-
Opticlean
-
-
-
-
Orientase 10B
-
-
-
-
P69 subtilase
-
-
Peptidase, subtilo-, A
-
-
-
-
phytophase
-
extensive homology to other plant subtilisin-like serine protease, phytaspase: a plant aspartate-specific protease
Protease S
-
-
-
-
Protease VIII
-
-
-
-
Protease XXVII
-
-
-
-
Proteinase K
-
-
Proteinase, Bacillus subtilis alkaline
-
-
-
-
Protin A 3L
-
-
-
-
psychrophilic subtilisin-like protease
-
-
S1P subtilase
-
-
SAS-1
-
extensive homology to other plant subtilisin-like serine protease
saspase
-
saspase: serine proteases exhibting aspase activity
Savinase
-
-
-
-
Savinase 16.0L
-
-
-
-
Savinase 32.0 L EX
-
-
-
-
Savinase 4.0T
-
-
-
-
Savinase 8.0L
-
-
-
-
SDD1 subtilase
-
-
senescence-associated subtilisin protease
-
SISBT3 subtilase
-
-
SP 266
-
-
-
-
SSU0757
-
gene name
subtilisin
-
-
subtilisin
synthetic construct
-
-
subtilisin 72
-
-
subtilisin 72
Bacillus subtilis 72
-
-
-
subtilisin A
-
Subtilisin amylosacchariticus
-
from Bacillus subtilis var. amylosacchariticus
Subtilisin amylosacchariticus
-
from Bacillus subtilis var. amylosacchariticus
Subtilisin amylosacchariticus
-
from Bacillus subtilis var. amylosacchariticus
Subtilisin BL
-
-
subtilisin Carlsberg
-
subtilisin Carlsberg
-
-
subtilisin Carlsberg
-
subtilisin Carlsberg
synthetic construct
-
-
subtilisin DJ-4
-
strain DJ-4
subtilisin DJ-4
-
strain DJ-4
-
Subtilisin DY
-
-
Subtilisin DY
-
-
Subtilisin DY
-
-
Subtilisin DY
-
-
Subtilisin DY
Bacillus subtilis DY
-
;
-
Subtilisin E
-
-
-
-
Subtilisin GX
-
-
-
-
subtilisin JB1
-
subtilisin JB1
Bacillus subtilis JB1
-
-
subtilisin Karlsberg
-
-
Subtilisin Novo
-
-
-
-
subtilisin Pr1-like protease
-
-
subtilisin protease
-
-
subtilisin QK
-
-
subtilisin QK
Bacillus subtilis QK02
-
-
-
Subtilisin S41
-
-
-
-
Subtilisin S41
-
-
Subtilisin S41
Bacillus subtilis TA41
-
-
-
subtilisin S4I
-
subtilisin S88
-
-
Subtilisin Sendai
-
-
-
-
subtilisin Sph
-
-
subtilisin-like protease
-
subtilisin-like protease
Alkalimonas collagenimarina AC40T
-
-
subtilisin-like protease
-
subtilisin-like protease
Dichelobacter nodosus C305
-
-
subtilisin-like protease
-
-
subtilisin-like protease
-
-
subtilisin-like protease
-
-
subtilisin-like protease
Fusarium equiseti CBS119568
-
-
-
subtilisin-like protease
-
-
subtilisin-like protease
-
-
subtilisin-like protease AprV2
-
-
subtilisin-like serine protease
-
subtilisin-like serine protease
-
subtilisin-like serine protease
-
-
subtilisin-like serine protease
-
subtilisin-like serine protease
-
subtilisin-like serine protease
-
subtilisin-like serine protease
-
subtilisin-like serine protease
C5VW36
-
subtilisin-like serine protease
C5VW36
-
-
subtilisin-like serine protease
-
-
subtilisin-like serine protease
-
subtilisin-like serine protease
-
subtilisin-like serine protease
Trichoderma harzianum ACCC30371
-
-
subtilisin-like serine protease
-
Subtilopeptidase
-
-
-
-
Superase
-
-
-
-
Thermoase
-
-
-
-
Thermoase PC 10
-
-
-
-
thermophilic thermitase
-
-
thermostable subtilisin
-
-
thermostable subtilisin
Bacillus sp. Ak.1
-
-
-
ThSS45
Trichoderma harzianum ACCC30371
gene name
-
trans-cinnamoyl-subtilisin
-
acyl-enzyme covalent intermediate of the serine protese subtilisin Carlsberg
mesenteroicopeptidase
-
-
additional information
-
species variants include subtilisin BPN' (also subtilisin B, subtilopeptidase B, subtilopeptidase C, Nagarse, Nagarse proteinase, subtilisin Novo, bacterial proteinase Novo) and subtilisin Carlsberg (subtilisin A, subtilopeptidase A, alcalase Novo), similar enzymes are produced by various Bacillus subtilis strains and other Bacillus species
additional information
-
synonyms of subtilisin Carlsberg: subtilisin, subtilisin A, subtilopeptidase A, alcalase Novo, synonyms of subtilisin BPN': nagarse proteinase, Nagarse, BPN', subtilopeptidase C, synonyms of subtilisin Novo: bacterial proteinase Novo, subtilisin B, subtilopeptidase B
additional information
-
species variants include subtilisin BPN' (also subtilisin B, subtilopeptidase B, subtilopeptidase C, Nagarse, Nagarse proteinase, subtilisin Novo, bacterial proteinase Novo) and subtilisin Carlsberg (subtilisin A, subtilopeptidase A, alcalase Novo), similar enzymes are produced by various Bacillus subtilis strains and other Bacillus species
additional information
-
synonyms of subtilisin Carlsberg: subtilisin, subtilisin A, subtilopeptidase A, alcalase Novo, synonyms of subtilisin BPN': nagarse proteinase, Nagarse, BPN', subtilopeptidase C, synonyms of subtilisin Novo: bacterial proteinase Novo, subtilisin B, subtilopeptidase B
additional information
-
the commercial enzyme subtilisin BPN' from Sigma: kinetic and physical properties indicate that the commercial enzyme is probably subtilisin Carlsberg isolated from Bacillus licheniformis, much work which has been performed on commercial subtilisin BPN' from 1975 up to 1985 must be revaluated
additional information
-
synonyms of subtilisin Carlsberg: subtilisin, subtilisin A, subtilopeptidase A, alcalase Novo, synonyms of subtilisin BPN': nagarse proteinase, Nagarse, BPN', subtilopeptidase C, synonyms of subtilisin Novo: bacterial proteinase Novo, subtilisin B, subtilopeptidase B
CAS REGISTRY NUMBER
COMMENTARY
9014-01-1
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Alkalimonas collagenimarina AC40T
strain AC40T
UniProt
Manually annotated by BRENDA team
Arabidopsis genome contains 56 subtilase genes
-
-
Manually annotated by BRENDA team
ecotype Col-0, gene At3g14067
UniProt
Manually annotated by BRENDA team
Bacillus alcalophilus Opticlean
Opticlean
-
-
Manually annotated by BRENDA team
double-cysteine mutants are constructed and recombinant plasmids are expressed in Bacillus subtilis
-
-
Manually annotated by BRENDA team
expression in Bacillus subtilis
-
-
Manually annotated by BRENDA team
expression of subtilisin BPN' gene isolated in a protease-deficient strain of Bacillus subtilis
-
-
Manually annotated by BRENDA team
recombinant protein
-
-
Manually annotated by BRENDA team
subtilisin BPN'; subtilisin Novo
-
-
Manually annotated by BRENDA team
wild-type and mutant (Asp to Ser99), the wild-type subtilisin BPN' is expressed and secreted by Bacillus subtilis DB104
-
-
Manually annotated by BRENDA team
wild-type and subtilisin 7150 (mutagenized subtilisin with enhanced thermal stability)
-
-
Manually annotated by BRENDA team
strain GMBAE 42, highest activity in late stationary growth phase in protein-rich medium at pH 10.5
-
-
Manually annotated by BRENDA team
Bacillus clausii GMBAE 42
strain GMBAE 42, highest activity in late stationary growth phase in protein-rich medium at pH 10.5
-
-
Manually annotated by BRENDA team
-
GN111900.1
GenBank
Manually annotated by BRENDA team
Bacillus gibsonii DSM 14391
-
GN111900.1
GenBank
Manually annotated by BRENDA team
strain 3-19
-
-
Manually annotated by BRENDA team
Bacillus intermedius Mrz 19
strain 3-19
-
-
Manually annotated by BRENDA team
isoforms BLAPP, BLAP X
-
-
Manually annotated by BRENDA team
Savinase TM
-
-
Manually annotated by BRENDA team
subtilisin BL cloned and overexpressed in Bacillus subtilis DB104
-
-
Manually annotated by BRENDA team
japanese strain DY
-
-
Manually annotated by BRENDA team
NCIB 6816; subtilisin Carlsberg
-
-
Manually annotated by BRENDA team
subtilisin Carlsberg
-
-
Manually annotated by BRENDA team
bacterial enzyme preparation, Alcalase from Novo Industries; crude enzyme preparation, Alcalase 2.0T from Novo; subtilisin Novo
-
-
Manually annotated by BRENDA team
commercial preparation from Genencor International, Purafect UF concentrate, Purafect 4000D, 4000E
-
-
Manually annotated by BRENDA team
for the commercial enzyme subtilsin BPN' from Sigma kinetic and physical properties indicate that the commercial enzyme is probably subtilisin Carlsberg isolated from Bacillus licheniformis, much work which has been performed on commercial subtilisin BPN' from 1975 up to 1985 must be revaluated
-
-
Manually annotated by BRENDA team
from Nagase Biochemicals
-
-
Manually annotated by BRENDA team
mutant Savinase; subtilisin Novo
-
-
Manually annotated by BRENDA team
proenzyme; strain DJ-4
SwissProt
Manually annotated by BRENDA team
Q103R subtilisin E
-
-
Manually annotated by BRENDA team
strain 1168
-
-
Manually annotated by BRENDA team
strain Ak.1
-
-
Manually annotated by BRENDA team
strain Ak.1
Uniprot
Manually annotated by BRENDA team
strain DJ-1
-
-
Manually annotated by BRENDA team
strain DJ-4, isolated from Korean fermented soybean food, Doen-Jang
-
-
Manually annotated by BRENDA team
strain G-825-6 (subtilisin Sendai)
-
-
Manually annotated by BRENDA team
strain GX6644 (subtilisin GX)
-
-
Manually annotated by BRENDA team
strain TA41 (antarctic psychrophilic Bacillus)
-
-
Manually annotated by BRENDA team
subtilisin amylosacchariticus
-
-
Manually annotated by BRENDA team
subtilisin amylosacchariticus; subtilisin DY
-
-
Manually annotated by BRENDA team
subtilisin amylosacchariticus; subtilisin Novo
-
-
Manually annotated by BRENDA team
subtilisin BPN' from Bacillus subtilis, the identity of this strain is later questioned and classified as a species of Bacillus amyloliquefaciens; subtilisin Novo; subtilopeptidase
-
-
Manually annotated by BRENDA team
subtilisin BPN' from Enzyme Development Corporation, New York
-
-
Manually annotated by BRENDA team
subtilisin BPN' from Serva; subtilisin DY; subtilisin from Boehringer
-
-
Manually annotated by BRENDA team
subtilisin BPN' from Sigma; subtilisin E; subtilisin from Sigma
-
-
Manually annotated by BRENDA team
subtilisin Carlsberg type VIII; subtilisin from Sigma
-
-
Manually annotated by BRENDA team
subtilisin DY
-
-
Manually annotated by BRENDA team
subtilisin DY; subtilisin DY, a natural mutant of subtilisin Carlsberg
-
-
Manually annotated by BRENDA team
subtilisin DY; subtilisin from Sigma
-
-
Manually annotated by BRENDA team
subtilisin E
-
-
Manually annotated by BRENDA team
subtilisin E; subtilisin E wild-type and mutant D32N produced in Escherichia coli
-
-
Manually annotated by BRENDA team
subtilisin from Novo Industries
-
-
Manually annotated by BRENDA team
subtilisin from Novo Industries; subtilisin from Sigma
-
-
Manually annotated by BRENDA team
subtilisin from Sigma
-
-
Manually annotated by BRENDA team
subtilisin Novo
-
-
Manually annotated by BRENDA team
subtilisin Novo; subtilisin Novo chemically attached to soluble DEAE-dextran and insoluble DEAE-Sephadex
-
-
Manually annotated by BRENDA team
Bacillus sp. 1168
strain 1168
-
-
Manually annotated by BRENDA team
Bacillus sp. Ak.1
strain Ak.1
-
-
Manually annotated by BRENDA team
Bacillus sp. Ak.1
strain Ak.1
Uniprot
Manually annotated by BRENDA team
strain DJ-1
-
-
Manually annotated by BRENDA team
proenzyme; strain DJ-4
SwissProt
Manually annotated by BRENDA team
strain DJ-4, isolated from Korean fermented soybean food, Doen-Jang
-
-
Manually annotated by BRENDA team
strain G-825-6 (subtilisin Sendai)
-
-
Manually annotated by BRENDA team
Bacillus sp. GX6644
strain GX6644 (subtilisin GX)
-
-
Manually annotated by BRENDA team
Bacillus sp. TA41
strain TA41 (antarctic psychrophilic Bacillus)
-
-
Manually annotated by BRENDA team
Bacillus subtilis enzyme, pro-subtilisin E produced in Escherichia coli
-
-
Manually annotated by BRENDA team
gene aprE
SwissProt
Manually annotated by BRENDA team
strain 1168
-
-
Manually annotated by BRENDA team
strain CN2, growth on soy peptone medium
-
-
Manually annotated by BRENDA team
strain DB104 harboring the subtilisin gene ligated into plasmid pUB110
-
-
Manually annotated by BRENDA team
strain DB428 a clone producing Q103R subtilisin E
-
-
Manually annotated by BRENDA team
strain DY; subtilisin DY (a natural mutant of subtilisin Carlsberg)
-
-
Manually annotated by BRENDA team
strain IBTC-3
-
-
Manually annotated by BRENDA team
strain RT-5, a thermostable soil isolate from the Tharparkar desert of Pakistan
-
-
Manually annotated by BRENDA team
strains BAL941, BAL1190 and BAL2838
-
-
Manually annotated by BRENDA team
subtilisin E
-
-
Manually annotated by BRENDA team
var. Carlsberg, commercial preparation, Novo Industrial, with spec. activity 2.5 AU per ml
-
-
Manually annotated by BRENDA team
Bacillus subtilis 1168
strain 1168
-
-
Manually annotated by BRENDA team
Bacillus subtilis 72
strain 72
-
-
Manually annotated by BRENDA team
Bacillus subtilis CN2
strain CN2, growth on soy peptone medium
-
-
Manually annotated by BRENDA team
Bacillus subtilis DB104
strain DB104 harboring the subtilisin gene ligated into plasmid pUB110
-
-
Manually annotated by BRENDA team
Bacillus subtilis DB428
strain DB428 a clone producing Q103R subtilisin E
-
-
Manually annotated by BRENDA team
Bacillus subtilis DY
strain DY
-
-
Manually annotated by BRENDA team
Bacillus subtilis IBTC-3
strain IBTC-3
-
-
Manually annotated by BRENDA team
Bacillus subtilis JB1
JB1
UniProt
Manually annotated by BRENDA team
Bacillus subtilis QK02
QK02
-
-
Manually annotated by BRENDA team
Bacillus subtilis RT-5
strain RT-5, a thermostable soil isolate from the Tharparkar desert of Pakistan
-
-
Manually annotated by BRENDA team
Bacillus subtilis TA41
-
-
-
Manually annotated by BRENDA team
AprV2; strain VCS1703A
UniProt
Manually annotated by BRENDA team
BprB; strain C305
UniProt
Manually annotated by BRENDA team
BprV; strain VCS1703A
UniProt
Manually annotated by BRENDA team
Dichelobacter nodosus C305
BprB; strain C305
UniProt
Manually annotated by BRENDA team
AprV2; strain VCS1703A
UniProt
Manually annotated by BRENDA team
BprV; strain VCS1703A
UniProt
Manually annotated by BRENDA team
collected at Base Frei Montalva, King George Island, Chilean Antarctic
-
-
Manually annotated by BRENDA team
gene prtS8A
-
-
Manually annotated by BRENDA team
Fusarium equiseti CBS119568
gene prtS8A
-
-
Manually annotated by BRENDA team
subtilisin J, Bacillus stearothermophilus NCIMB10278 gene expressed in Bacillus subtilis DB104/pZS101
-
-
Manually annotated by BRENDA team
strains GS115 and KM71
-
-
Manually annotated by BRENDA team
KV71, KV22 and KV42
-
-
Manually annotated by BRENDA team
KV01 and KV54
-
-
Manually annotated by BRENDA team
an avirulent isolate
-
-
Manually annotated by BRENDA team
strain P1/7
C5VW36
UniProt
Manually annotated by BRENDA team
strain P1/7
C5VW36
UniProt
Manually annotated by BRENDA team
synthetic construct
-
-
-
Manually annotated by BRENDA team
gene ThSS45
UniProt
Manually annotated by BRENDA team
Trichoderma harzianum ACCC30371
gene ThSS45
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
the subtilisin-like proteases share the same catalytic mechanism as the trypsin-like proteases, depending upon the hydroxyl group of a serine residue. The catalytic triad of subtilisin-like proteases is composed of Asp, His, and Ser. Due to its different specificity compared to the members of the S8 family of clan SB of proteases, the Fe protease might be a protease distinct from previously defined IUBMB groups of proteases, it is no member of the the S8 peptidase family
evolution
-
the enzyme belongs to a structurally distinct class of the subtilase family
evolution
phylogenetic analysis, AtSASP and its putative orthologues clustering in one discrete group of subtilisin proteases in which no other Arabidospsis subtilisin protease is present. The enzyme function is at least partially conserved between Arabidopsis thaliana and Oryza sativa
evolution
the enzyme is a member of the peptidases_S8/PCSK9/proteinase K-like family (Pf00082)
evolution
Fusarium equiseti CBS119568
-
the subtilisin-like proteases share the same catalytic mechanism as the trypsin-like proteases, depending upon the hydroxyl group of a serine residue. The catalytic triad of subtilisin-like proteases is composed of Asp, His, and Ser. Due to its different specificity compared to the members of the S8 family of clan SB of proteases, the Fe protease might be a protease distinct from previously defined IUBMB groups of proteases, it is no member of the the S8 peptidase family
-
evolution
Trichoderma harzianum ACCC30371
-
the enzyme is a member of the peptidases_S8/PCSK9/proteinase K-like family (Pf00082)
-
malfunction
-
gene knock-out results in reduced ability by the parasite to undergo promastigote to amastigote differentiation in vitro. SUB-deficient Leishmania display reduced virulence in infection models. SUB knock-out parasites show altered regulation of the terminal peroxidases of the trypanothione reductase system and the predominant tryparedoxin peroxidases are decreased in SUB-/- parasites. knock-out parasites show increased sensitivity to hydroperoxide. Data suggest that subtilisin is the maturase for tryparedoxin peroxidases and is necessary for full virulence
malfunction
-
using a protease mutant of Dichelobacter nodosus it is shown in a sheep virulence model that AprV2 is required for virulence
physiological function
strains causing benign footrot secrete AprB2; strains causing benign footrot secrete subtilisin-like protease BprB; strains causing virulent footrot secrete the subtilisin-like protease AprV2; strains causing virulent footrot secrete the subtilisin-like protease BprV
physiological function
-
during growth of Pichia pastoris, Sub2 is produced as a secreted enzyme at a concentration of 10 microg/ml of culture supernatant after overexpression of the full-length SUB2 gene. During fermentative production of recombinant enzymes in methanol medium, 1 ml of Pichia pastoris culture supernatant is found to contain approximately 3 ng of Sub2, while the enzyme is not detected during growth in a medium containing glycerol as a carbon source
physiological function
AcpII comprises a prepropeptide, a catalytic domain that includes a protease-associated domain, and tandem repeat prepeptidase C-terminal domains
physiological function
synthetic construct
-
subtilisin enhances transpeptidase activity of 67 kDa gamma-glutamyl transpeptidase and 30 kDa gamma-glutamyl transpeptidase, by nearly 1.5- and 2fold, respectively. In presence of subtilisin, 30 kDa gamma-glutamyl transpeptidase has improved catalytic efficiency, altered pH and temperature optima and has salt-tolerant glutaminase activity
physiological function
subtilisin-like serine proteases from nematode-trapping fungi are involved in the penetration and digestion of nematode cuticles
physiological function
subtilisin-like serine proteases from nematode-trapping fungi are involved in the penetration and digestion of nematode cuticles
physiological function
subtilisin-like serine proteases from nematode-trapping fungi are involved in the penetration and digestion of nematode cuticles
physiological function
subtilisin-like serine proteases from nematode-trapping fungi are involved in the penetration and digestion of nematode cuticles
physiological function
subtilisin-like serine proteases from nematode-trapping fungi are involved in the penetration and digestion of nematode cuticles
physiological function
subtilisin-like serine proteases from nematode-trapping fungi are involved in the penetration and digestion of nematode cuticles
physiological function
C5VW36
is involved in Streptococcus suis virulence
physiological function
synthetic construct
-
peanut hydrolysate obtained after 6 h of digestion by alcalase used to isolate angiotensin I converting enzyme inhibitory peptides
physiological function
-
silk fibroin hydrolysate obtained through alcalase digestion demonstrates in vitro angiotensin I converting enzyme inhibitory activity and in vivo antihypertensive activity in spontaneously hypertensive rats. Degrees of hydrolysis and angiotensin I converting enzyme inhibitory activities is related with hydrolysis period of silk fibroin hydrolysates obtained with alcalase treatments: highest angiotensin I converting enzyme inhibitory activities (72.5%) are obtained with a hydrolysis time of 105 min, whereby degree of hydrolysis is 17.1%
physiological function
-
isolation of angiotensin I-converting enzyme inhibitory peptide from hydrolysates and enzymatic digests of Spirulina platensis by alcalase. The digests by alcalase with a molecular weight range of 0-3000 show the most potent inhibitory activity of 0.23 mg/ml with a yield of 15.0% among tested hydrolysates
physiological function
-
when subtilisin C. is chemically modified with polyethylene glycol and inhibited with a dansyl fluorophore, and initially dissolved in two organic solvents (acetonitrile and 1,4-dioxane), the active site environment of the enzyme is similar to that in water. Prolonged exposure to the organic medium causes this environment to resemble that of the solvent in which the enzyme is dissolved
physiological function
-
significantly inhibits carrageenan-induced mouse tail thrombus formation in vivo. Inhibition activity increases along with the injection amount of subtilisin QK, which presents linearity relationship. When the amount of subtilisin QK reaches 12000 IU, thrombus nearly disappears in mouse tail
physiological function
-
alcalase-hydrolyzed potato protein has both antioxidant and emulsifying properties
physiological function
-
Pro-subtilisin-like serine protease consists of an N-terminal propeptide (Ala1-Ala113), a mature domain (subtilisin-like serine protease, Val114-Val539) and a C-terminal propeptide (Asp540-Gly640)
physiological function
-
acts extracellularly in the apoplast of stomatal precursor cells where it may be involved in the generation of signals responsible for stomata density regulation; implicated in fragmentation of the membrane-bound transcription factor AtbZIP1723 and precursors of pectin methylesterase and rapid alkalinization growth factor; the subtilase ALE1 is involved in the generation of peptide signals that are required for cuticle formation and epidermal differentiation during Arabidopsis embryo development
physiological function
-
some caspase-like activities are attributable to the plant subtilisin-like proteases, saspases and phytaspases. Enzyme hydrolyzes a range of tetrapeptide caspase substrates following the aspartate residue. Enzyme is implicated the proteolytic degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) during biotic and abiotic programmed cell death
physiological function
-
involved in proteolysis of the seed storage protein, beta-conglycinin
physiological function
-
enzyme shows caspase specificity; some caspase-like activities are attributable to the plant subtilisin-like proteases, saspases and phytaspases. Enzyme hydrolyzes a range of tetrapeptide caspase substrates following the aspartate residue. In response to death-inducing stimuli, phytaspase is shown to re-localize to the cell interior
physiological function
-
pathogenesis related protein which is shown to be one of several subtilases that are specifically induced following pathogen infection. P69 is suggested to process a leucin-rich repeat cell wall protein in virus-infected tomato plants
physiological function
-
principal allergen produced by Aspergillus strains
physiological function
-
Subtilisin A applied at nanomolar concentrations suppresses epileptiform spikes in rat hippocampal slices and neocortex in vivo
physiological function
-
modeling of intracellular subtilisin protease regulation within the cell
physiological function
-
a cold-adapted protease, residue Ala284 is an important cold-adaptation determinant of the enzyme
physiological function
-
fungal subtilisin is a defense elicitor, culture filtrates from avirulent Acremonium spp. induce resistance against pathogens in strawberry plants or accumulation of reactive oxygen species (e.g. H2O2 and O2 -) and callose in Arabidopsis thaliana plants, expression analysis of defense-related genes in strawberry and defense responses induced by the enzyme in Arabidopsis leaves, overview
physiological function
the enzyme downregulates branching and silique production during monocarpic senescence
physiological function
the serine protease may be involved in both mycoparasitism and antibiotic secretion
physiological function
Alkalimonas collagenimarina AC40T
-
AcpII comprises a prepropeptide, a catalytic domain that includes a protease-associated domain, and tandem repeat prepeptidase C-terminal domains
-
physiological function
Bacillus subtilis QK02
-
significantly inhibits carrageenan-induced mouse tail thrombus formation in vivo. Inhibition activity increases along with the injection amount of subtilisin QK, which presents linearity relationship. When the amount of subtilisin QK reaches 12000 IU, thrombus nearly disappears in mouse tail
-
physiological function
Dichelobacter nodosus C305
-
strains causing benign footrot secrete subtilisin-like protease BprB
-
physiological function
-
strains causing virulent footrot secrete the subtilisin-like protease AprV2; strains causing virulent footrot secrete the subtilisin-like protease BprV
-
physiological function
-
is involved in Streptococcus suis virulence
-
physiological function
Trichoderma harzianum ACCC30371
-
the serine protease may be involved in both mycoparasitism and antibiotic secretion
-
malfunction
at maturity, enzyme knockout sasp-1 plants produce 25% more inflorescence branches and siliques, mostly due to an increased number of second and third order branches, than either the wild-type or the rescued lines
additional information
-
the catalytic triad residues consists of Ser221, Pro210 to His64
additional information
-
structure molecular modeling using the crystal structure of the pro-subtilisin E complex, PDB ID 1SCJ, the active site Cys221 is replaced with the catalytic serine, molecular dynamics simulation, overview
additional information
-
substrate specificity and the role of stress signals such as divalent metal ions play roles in defining the proteolytic activity of Bacillus clausii intracellular subtilisin protease, molecular basis, overview. Heat-denatured whole proteins are found to be better substrates for the enzyme than the native forms. The S1, S2 and S4 sites form defined substrate binding pockets
additional information
-
structure homology modeling, structure comparisons, overview
additional information
-
molecular modeling and docking analysis using crystal structure PDB ID 1yu6
additional information
occurrence of the two variant forms of AtSASP can be due to posttranslational modifications
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R,S)-2-benzyl-3-(tert-butylsulfonyl)propionic acid + H2O
(R)-2-benzyl-3-(tert-butylsulfonyl)propionic ethyl ester + (S)-2-benzyl-3-(tert-butylsulfonyl)propionic acid + ethanol
show the reaction diagram
-
-
-
-
(R,S)-2-benzyl-3-[[1-methyl-1-((morpholin-4-yl)-carbonyl)ethyl]sulfonyl]propionic acid + H2O
(S)-2-benzyl-3-[[1-methyl-1-((morpholin-4-yl)-carbonyl)ethyl]sulfonyl]propionic acid + (R)-2-benzyl-3-[[1-methyl-1-((morpholin-4-yl)-carbonyl)ethyl]sulfonyl]propionic acid-ethyl ester + ethanol
show the reaction diagram
-
-
-
-
(R/S)-(2-methylpropyl)butanedioic acid diethyl ester + H2O
(S)-(2-methylpropyl)butanedioic acid diethyl ester + (R)-(2-methylpropyl)butanedioic acid 4-ethyl ester + ethanol
show the reaction diagram
-
-
-
-
67 kDa gamma-glutamyl transpeptidase + H2O
30 kDa gamma-glutamyl transpeptidase + ?
show the reaction diagram
synthetic construct
-
proteolytic digestion of 67 kDa gamma-glutamyl transpeptidase from Bacillus licheniformis ER-15 by subtilisin to the 30 kDa form, which in turn remains associated with subtilisin as a heterodimeric protein
-
?
Aalpha chain of fibrinogen + H2O
?
show the reaction diagram
-
-
-
-
?
acetyl-L-Ala-L-Ala-L-Ala-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
acetyl-L-Phe + ethanol
acetyl-L-Phe ethyl ester + H2O
show the reaction diagram
Bacillus subtilis, Bacillus subtilis IBTC-3
-
-
-
r
acetyl-L-Phe ethyl ester + H2O
acetyl-L-Phe + ethanol
show the reaction diagram
Bacillus subtilis, Bacillus subtilis IBTC-3
-
-
-
r
acetyl-L-Tyr + ethanol
acetyl-L-Tyr ethyl ester + H2O
show the reaction diagram
-
-
-
r
acetyl-L-Tyr ethyl ester + H2O
acetyl-L-Tyr + ethanol
show the reaction diagram
Bacillus subtilis, Bacillus subtilis IBTC-3
-
-
-
r
acid casein + H2O
?
show the reaction diagram
-
purified caseins from animal's milk from cow, sheep, goat and water buffalo used as substrate for subtilisin, cow acid casein is the best substrate
-
-
?
Ala-Ala-Phe 7-amido-4-methyl coumarin + H2O
?
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe 7-amido-4-methyl coumarin + H2O
?
show the reaction diagram
Bacillus subtilis, Bacillus subtilis JB1
-
-
-
?
Ala-Ala-Pro-Leu-4-nitroanilide + H2O
Ala-Ala-Pro-Leu + 4-nitroaniline
show the reaction diagram
-
low activity
-
?
Ala-Ala-Pro-Lys-4-nitroanilide + H2O
Ala-Ala-Pro-Lys + 4-nitroaniline
show the reaction diagram
-
low activity
-
?
Ala-Ala-Pro-Met-4-nitroanilide + H2O
Ala-Ala-Pro-Met + 4-nitroaniline
show the reaction diagram
-
low activity
-
?
Ala-Ala-Pro-Nle-4-nitroanilide + H2O
Ala-Ala-Pro-Nle + 4-nitroaniline
show the reaction diagram
-
low activity
-
?
Ala-Ala-Pro-Phe 4-nitroanilide + H2O
?
show the reaction diagram
Bacillus subtilis, Bacillus subtilis DB428
-
-
-
-
-
Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
low activity
-
?
Ala-Ala-Val-Ala-4-nitroanilide + H2O
Ala-Ala-Val-Ala + 4-nitroaniline
show the reaction diagram
-
low activity
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
azocasein + H2O
?
show the reaction diagram
Bacillus subtilis, Bacillus subtilis JB1
-
-
-
?
Benzoyl-Arg ethyl ester + H2O
?
show the reaction diagram
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
Benzoyl-Arg ethyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN', subtilisin Amylosacchariticus
-
-
-
benzoyl-L-Arg + ethanol
benzoyl-L-Arg ethyl ester + H2O
show the reaction diagram
-
-
-
r
Benzoyl-L-Arg ethyl ester + H2O
Benzoyl-L-Arg + ethanol
show the reaction diagram
Bacillus subtilis, Bacillus subtilis IBTC-3
-
-
-
r
Benzyloxycarbonyl-Ala-Ala-Leu 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly-Gly-L-Leu-NH2 + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly-L-Leu-NH2 + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly-L-Tyr-NH2 + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-glycyl-L-tyrosinamide + H2O
?
show the reaction diagram
-
-
-
-
-
benzyloxycarbonyl-L-Ala ethyl ester + butanol
benzyloxycarbonyl-L-Ala butyl ester + ethanol
show the reaction diagram
-
in isooctane, preferential use of L-enantiomer. Comparison with enantioselectivity in water and DMSO
-
?
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-4-nitroanilide + H2O
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu + 4-nitroaniline
show the reaction diagram
Bacillus intermedius, Bacillus intermedius Mrz 19
-
-
-
?
benzyloxycarbonyl-L-Asp methyl ester + serine amide
benzyloxycarbonyl-L-Asp-L-Ser-NH2 + methanol
show the reaction diagram
-
-
-
?
beta subunit of haemoglobin + H2O
?
show the reaction diagram
-
-
-
-
?
beta-casein + H2O
?
show the reaction diagram
Fusarium equiseti, Fusarium equiseti CBS119568
-
from bovine milk
-
-
?
Bovine serum albumin + H2O
?
show the reaction diagram
-
-
-
-
?
Bovine serum albumin + H2O
?
show the reaction diagram
-
-
-
-
?
Bovine serum albumin + H2O
?
show the reaction diagram
-
-
-
-
?
Bovine serum albumin + H2O
?
show the reaction diagram
Bacillus subtilis, Bacillus subtilis JB1
-
-
-
?
bovine serum albumin+ H2O
?
show the reaction diagram
-
poor substrate in native form, but can be digested in heat-denatured form
-
-
?
Carboxybenzoyl-Gly-Ala-NH2 + H2O
Carboxybenzoyl-Gly-Ala + NH3
show the reaction diagram
-
subtilisin BPN'
subtilisin BPN'
-
Carboxybenzoyl-Gly-Leu-NH2 + H2O
Carboxybenzoyl-Gly-Leu + NH3
show the reaction diagram
-
subtilisin BPN'
subtilisin BPN'
-
Carboxybenzoyl-Gly-Pro-Leu-Gly-Pro-OH + H2O
Carboxybenzoyl-Gly-Pro-Leu + Gly-Pro-OH
show the reaction diagram
-
subtilisin BPN'
subtilisin BPN'
-
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
-
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
38% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on casein
-
-
?
casein + H2O
?
show the reaction diagram
-
caseinolytic activity is determined using the casein plate method
-
-
?
casein + H2O
?
show the reaction diagram
Bacillus intermedius Mrz 19
-
-
-
-
?
casein + H2O
?
show the reaction diagram
Bacillus subtilis CN2
-
-
-
-
?
casein + H2O
?
show the reaction diagram
Alkalimonas collagenimarina AC40T
38% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on casein
-
-
?
chicken-feather keratin + H2O
?
show the reaction diagram
-
-
-
-
?
Clupein + H2O
?
show the reaction diagram
-
-
-
-
-
Collagen + H2O
?
show the reaction diagram
Alkalimonas collagenimarina, Alkalimonas collagenimarina AC40T
81% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on collagen
-
-
?
cuticle + H2O
?
show the reaction diagram
-
Pr1-like activity on host and non-host cuticle. In KV01 cultures, at 16 h, similar Pr1-like activity on both aphid and locust cuticle
-
-
?
cuticle + H2O
?
show the reaction diagram
-
Pr1-like activity on host and non-host cuticle. In KV71 cultures, at 16 and 24 h, similar Pr1-like activity on both aphid and locust cuticle. Filtrates from KV71 and KV42 cultures, that contain cocktails of cuticle-degrading enzymes, hydrolyse 3-4fold more protein from aphid than locust cuticle
-
-
?
Cytochrome c + H2O
?
show the reaction diagram
Fusarium equiseti, Fusarium equiseti CBS119568
-
from equine heart
-
-
?
D-Val-Leu-Lys p-nitroanilide + H2O
D-Val-Leu-Lys + p-nitroaniline
show the reaction diagram
-
-
?
eglin-c + H2O
?
show the reaction diagram
-
proteinase inhibitor, subtilisin DY hydrolyses the peptide bond between residues 45 and 46 in the reactive centre of eglin-c
-
-
?
Elastin + H2O
?
show the reaction diagram
-
-
-
-
?
Elastin + H2O
?
show the reaction diagram
7% activity relative to gelatin as substrate
-
-
?
ethyl 2-(4-ethylphenoxy)propionate + butanol
butyl 2-(4-ethylphenoxy)propanoate + propanol
show the reaction diagram
-
in isooctane, preferential use of S-enantiomer. Comparison with enantioselectivity in water and DMSO
-
?
Fibrin + H2O
?
show the reaction diagram
-
-
-
-
?
Fibrin + H2O
?
show the reaction diagram
-
strain DJ-4
-
-
?
Fibrin + H2O
?
show the reaction diagram
Bacillus subtilis QK02
-
-
-
-
?
Fibrin + H2O
?
show the reaction diagram
-
strain DJ-4
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
-
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
rapid hydrolysis of Aalpha-, Bbeta- and gamma-chains. At very low concentration, no cleavage of gamma-chain
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
Bacillus subtilis JB1
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
Alkalimonas collagenimarina, Alkalimonas collagenimarina AC40T
best substrate for AcpII. Also purified AcpII-C and AcpII-C-DELTAPA act well on gelatin
-
-
?
Gelatin + H2O
?
show the reaction diagram
Bacillus subtilis JB1
-
-
-
?
Glp-Ala-Ala-Leu-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
glutaryl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O
glutaryl-Ala-Ala-Pro-Leu + p-nitroaniline
show the reaction diagram
relative hydrolysis rate is 22%
-
?
Hammarsten casein + H2O
?
show the reaction diagram
Bacillus clausii, Bacillus clausii GMBAE 42
-
activation energy for hydrolysis 10.59 kcal per mol
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
-
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
-
-
-
-
Hemoglobin + H2O
?
show the reaction diagram
-
-
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
denatured
-
-
-
Human fibrinogen + H2O
?
show the reaction diagram
-
fibrinolytic activity is determined using the fibrin plate method
-
-
?
human fibronectin + H2O
?
show the reaction diagram
-
-
-
-
?
insoluble elastin + H2O
?
show the reaction diagram
-
-
-
-
?
kappa casein + H2O
?
show the reaction diagram
-
purified caseins from animal's milk from cow, sheep, goat and water buffalo used as substrate for subtilisin
-
-
?
Keratin + H2O
?
show the reaction diagram
-
-
-
-
?
Keratin + H2O
?
show the reaction diagram
Alkalimonas collagenimarina, Alkalimonas collagenimarina AC40T
1.5% activity relative to gelatin as substrate
-
-
?
L-Ala-L-Ala-L-Ala-L-Ala-L-Pro-L-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
L-Ala-L-Ala-L-Phe-L-Ala-L-Ala-L-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
L-Ala-L-Ala-L-Pro-L-Ala + H2O
?
show the reaction diagram
-
-
-
-
?
L-Ala-L-Ala-L-Pro-L-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
L-Ala-L-Ala-L-Val-L-Ala + H2O
?
show the reaction diagram
-
-
-
-
?
L-Lys-L-His-L-Asp-L-Arg-L-Lys-L-Asp + H2O
?
show the reaction diagram
-
-
-
-
?
L-Phe-L-Ala-L-Ala-L-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
L-phenylalanine-isopropylester + H2O
L-phenylalanine + isopropanol
show the reaction diagram
-
-
-
-
L-Tyr-L-Val-L-Ala-L-Asp + H2O
?
show the reaction diagram
-
-
-
-
?
lactate dehydrogenase + H2O
?
show the reaction diagram
-
poor substrate in native form, but can be digested in heat-denatured form
-
-
?
malate dehydrogenase + H2O
?
show the reaction diagram
-
poor substrate in native form, but can be digested in heat-denatured form
-
-
?
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
methoxysuccinyl-Ala-Ile-Pro-Met-p-nitroanilide + H2O
methoxysuccinyl-Ala-Ile-Pro-Met + p-nitroaniline
show the reaction diagram
is the most favorable substrate, relative hydrolysis rate is 100%
-
?
N-Acetyl-Ala methyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-Acetyl-Gly ethyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-Acetyl-L-norvaline ethyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-acetyl-L-phenylalanine ethyl ester
?
show the reaction diagram
-
-
-
-
?
N-acetyl-L-phenylalanine ethyl ester + 1-butanol
?
show the reaction diagram
-
transesterification catalyzed by poly(ethylene glycol)-modified subtilisin
-
-
?
N-acetyl-L-phenylalanine ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N-Acetyl-Leu methyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-Acetyl-Lys methyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-Acetyl-Phe ethyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-Acetyl-Phe methyl ester + H2O
?
show the reaction diagram
-
subtilisin Amylosacchariticus
-
-
-
N-Acetyl-Phe methyl ester + H2O
?
show the reaction diagram
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Trp ethyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-Acetyl-Trp methyl ester + H2O
?
show the reaction diagram
-
subtilisin Amylosacchariticus
-
-
-
N-Acetyl-Trp methyl ester + H2O
?
show the reaction diagram
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Tyr ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
N-Acetyl-Tyr ethyl ester + H2O
?
show the reaction diagram
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Tyr ethyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-acetyl-Tyr ethyl ester + H2O
N-acetyl-Tyr + ethanol
show the reaction diagram
-
-
?
N-Acetyl-Tyr methyl ester + H2O
?
show the reaction diagram
-
subtilisin Amylosacchariticus
-
-
-
N-Acetyl-Tyr methyl ester + H2O
?
show the reaction diagram
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Val methyl ester + H2O
?
show the reaction diagram
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Val methyl ester + H2O
?
show the reaction diagram
-
subtilisin BPN'
-
-
-
N-acetylglucosamine + H2O
?
show the reaction diagram
-
N-acetylglucosamine has little effect on subtilisin production by KV01 (11% of that on cuticle)
-
-
?
N-acetylglucosamine + H2O
?
show the reaction diagram
-
restricted feeding of N-acetylglucosamine (the monomer of chitin) to biomass of KV71 induces protease to 61% of that on cuticle
-
-
?
N-CBZ-Gly-Gly-Leu p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide + H2O
N-methoxysuccinyl-Ala-Ala-Pro-Val + 4-nitroanilide
show the reaction diagram
-
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide + H2O
N-methoxysuccinyl-Ala-Ala-Pro-Val + p-nitroaniline
show the reaction diagram
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide + H2O
N-methoxysuccinyl-Ala-Ala-Pro-Val + p-nitroaniline
show the reaction diagram
Dichelobacter nodosus C305
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide + H2O
N-methoxysuccinyl-Ala-Ala-Pro-Val + p-nitroaniline
show the reaction diagram
-
-
?
N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
N-Succ-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
?
N-succinyl-Ala-Ala-Pro-Leu-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Leu + 4-nitroaniline
show the reaction diagram
-
-
?
N-succinyl-Ala-Ala-Pro-Met-p-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Met + p-nitroaniline
show the reaction diagram
relative hydrolysis rate is 39%
-
?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
show the reaction diagram
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
show the reaction diagram
relative hydrolysis rate is 94%
-
?
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-succinyl-Ala-Ala-Val-Ala-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Val-Ala + 4-nitroaniline
show the reaction diagram
-
-
?
N-succinyl-L-Ala-L-Ala-L-L-Ala-4-nitroanilide + H2O
?
show the reaction diagram
Bacillus clausii, Bacillus clausii GMBAE 42
-
3.8% of the activity with N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide
-
-
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
N-succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide + H2O
?
show the reaction diagram
Bacillus clausii, Bacillus clausii GMBAE 42
-
19% of the activity with N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide
-
-
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
show the reaction diagram
Bacillus clausii GMBAE 42
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-Val-4-nitroanilide + H2O
?
show the reaction diagram
Bacillus clausii, Bacillus clausii GMBAE 42
-
2.2% of the activity with N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide
-
-
?
N-tosyl-L-Arg methyl ester + H2O
N-tosyl-L-Arg + methanol
show the reaction diagram
-
-
-
-
N-tosyl-L-Arg methyl ester + H2O
N-tosyl-L-Arg + methanol
show the reaction diagram
-
subtilisin Carlsberg, subtilisin Novo, subtilisin Amylosacchariticus
-
-
N-trans-cinnamoyl imidazole + H2O
(E)-cinnamate + imidazole
show the reaction diagram
-
-
-
?
Nalpha-benzoyl-DL-Arg-4-nitroanilide + H2O
Nalpha-benzoyl-DL-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
?
Nalpha-Benzoyl-L-Arg ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
Oxidized insulin B-chain + H2O
?
show the reaction diagram
-
enzyme primarily hydrolyzes Leu15-Tyr16 bond and secondarily Gln4-His5, Ser9-His10, Phe24-Phe25 and Lys29-Ala30
-
-
-
oxidized insulin chain B + H2O
?
show the reaction diagram
-
is cleaved by subtilisin-like serine protease at multiple sites, preferably at the C-termini of the hydrophobic residues, such as Tyr, Phe, Leu, Val and Ala. This peptide is also cleaved by subtilisin at the C-termini of the hydrophobic residues, but at more specific sites
-
-
?
p-nitrophenyl butyrate + H2O
p-nitrophenol
show the reaction diagram
-
catalyzed by native subtilisin
-
?
peanut flour + H2O
?
show the reaction diagram
synthetic construct
-
6 h alcalase hydrolysate
-
-
?
peptidyl-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
Phe-Ala-Ala-Phe-4-nitroanilide + H2O
Phe-Ala-Ala-Phe + 4-nitroaniline
show the reaction diagram
-
highly preferred substrate
-
?
porcine brain tubulin + H2O
peptides
show the reaction diagram
-
MALDI-mass spectrum of the carboxy-terminal peptides released by subtilisin treatment. Polyglutamylated peptides from the beta subunit and from both the tyrosinated and detyrosinated forms of the alpha subunit are observed. The fragments removed by subtilisin are in the range of 1.6 kDa. Post-translational modifications can be directly identified in the mixture of peptides resulting from limited subtilisin treatment
-
?
proPhrA + H2O
PhrA
show the reaction diagram
-
-
-
?
sec-phenethyl alcohol + vinyl butyrate
?
show the reaction diagram
-
-
-
-
?
serum albumin + H2O
?
show the reaction diagram
-
-
-
-
?
silk fibroin + H2O
?
show the reaction diagram
-
-
-
-
?
Spirulina platensis powder + H2O
?
show the reaction diagram
-
-
-
-
?
strobilurin fungicide + H2O
?
show the reaction diagram
-
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-methyl-coumaryl-7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
synthetic construct
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
Fusarium equiseti CBS119568
-
-
-
?
succinyl-AAPA-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
succinyl-AAPA-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
succinyl-AAPE-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
show the reaction diagram
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
show the reaction diagram
Bacillus sp. Ak.1
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-alanyl-alanyl propyl-phenylalanine
show the reaction diagram
-
-
-
?
succinyl-AAPR-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
Succinyl-Ala-Ala 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Succinyl-Ala-Ala 4-nitroanilide + H2O
?
show the reaction diagram
Bacillus sp. Ak.1
-
-
-
-
-
Succinyl-Ala-Ala-Ala 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Succinyl-Ala-Ala-Ala 4-nitroanilide + H2O
?
show the reaction diagram
Bacillus sp. Ak.1
-
-
-
-
-
succinyl-Ala-Ala-Ala-Ala-Ala-p-nitroanilide + H2O
succinyl-AAAAA + p-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-Ala-Ala-Ala-p-nitroanilide + H2O
succinyl-AAA + p-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-Ala-Ala-p-nitroanilide + H2O
succinyl-Ala-Ala + p-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-Ala-Ala-Pro-Ala-p-nitroanilide + H2O
succinyl-AAPA + p-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O
succinyl-AAPL + p-nitroaniline
show the reaction diagram
Bacillus sp., Bacillus sp. Ak.1
-
-
-
?
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
Bacillus subtilis DB428
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
Bacillus subtilis RT-5
-
-
-
-
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-Ala-Ala-Pro-Phe-7-amido-4-methylcoumarin + H2O
succinyl-Ala-Ala-Pro-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-Ala2-Phe-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Ala-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
succinyl-L-Ala-L-Ala-L-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-Gly-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-Gly + 4-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Ala-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Ala + 4-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Lys-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Lys + 4-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Met-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Met + 4-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Tyr-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Tyr + 4-nitroaniline
show the reaction diagram
-
-
-
?
succinyl-L-Asp-L-Val-L-Arg-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
succinyl-L-Asp-L-Val-L-Arg-L-Ala-L-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
succinyl-Phe-Ala-Ala-Phe-p-nitroanilide + H2O
succinyl-Phe-Ala-Ala-Phe + p-nitroaniline
show the reaction diagram
-
-
-
?
Toluenesulfonyl-Arg methyl ester + H2O
?
show the reaction diagram
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
type I keratin + H2O
?
show the reaction diagram
-
-
-
-
?
ubiquitin + H2O
?
show the reaction diagram
Fusarium equiseti, Fusarium equiseti CBS119568
-
from bovine blood cells
-
-
?
Urea-denatured hemoglobin + H2O
?
show the reaction diagram
Bacillus subtilis, Bacillus subtilis IBTC-3
-
-
-
-
?
vinyl butyrate + H2O
?
show the reaction diagram
-
-
-
-
?
Z-Ala-Ala-Leu-OCH3 + H2O
?
show the reaction diagram
-
-
-
-
?
Z-Ala-Ala-Leu-OMe + Phe-p-nitroanilide
Z-Ala-Ala-Leu-Phe-p-nitroanilide + methanol
show the reaction diagram
-
-
-
?
Z-Ala-Ala-Leu-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
Z-Ala-Phe-OMe + H2O
Z-Ala-Phe-NH2 + methanol
show the reaction diagram
-
enzymatic method for the synthesis of free terminal amides of peptides, by ammonolysis of peptide methyl esters using ammonium carbamate and subtilisin A from Bacillus licheniformis in polar organic solvents with low water content is developed. Enzyme is very stable and active in a mixture of t-BuOH and DMF 82.5:17.5 (v/v), containing 0.2% water. Optimum conditions for Z-Ala-Phe-NH2 synthesis are molar ratio ammonium carbamate to Z-Ala-Phe-OMe 10, in t-BuOH/DMF, 82.5:17.5 (v/v) containing 0.2% (v) water, at 30C for 21 h with the maximum yield of 87%
-
?
Z-Thr-Ala-Thr-OCH3 + Asp-p-nitroanilide
Z-Thr-Ala-Thr-Asp-p-nitroanilide + methanol
show the reaction diagram
-
-
-
?
methyl mandelate + butanol
butyl mandelate + methanol
show the reaction diagram
-
in isooctane, preferential use of S-enantiomer. Comparison with enantioselectivity in water and DMSO
-
?
additional information
?
-
-
random mutagenesis to enhance the activity of subtilisin in organic solvents
-
-
-
additional information
?
-
-
influence of substrate structure of N-protected peptide nitroanilides of the types: benzyloxycarbonyl-A2-A1 4-nitroanilide, benzyloxycarbonyl-A3-A2-A1 4-nitroanilide, benzyloxycarbonyl-A4-A3-A2-A1 4-nitroanilide, subsite S1 is of broad selectivity: preference for hydrophobic amino acid residues (i.e. leucine and phenylalanine) the beta-branched and the basic amino acid residues cannot interact with the S1 subsite and the hydrolysis of the corresponding peptides occurs exclusively at the A2-A1 bond. If S1/A1 interactions are weak (Ala, norvaline, norleucine) the amino acid residue A1 can interact with subsites S1 and S1' resulting in the hydrolysis of two bonds (A1 4-nitroanilide and A2-A1). The subsite S2 reveals a preference for small amino acid residues
-
-
-
additional information
?
-
-
under relatively non-aqueous conditions, immobilized subtilisin is able to synthesize phenylacetic acid ethyl ester
-
-
-
additional information
?
-
-
high elastolytic acitivity, preferentially hydrolyzes the ester bond of Ala, but significant hydrolysis is observed with other aliphatic (Gly, Leu) and aromatic (Tyr) amino acids
-
-
-
additional information
?
-
-
reactions catalyzed: 1. peptide bond hydrolysis, 2. ester bond hydrolysis, 3. transesterification, 4. transpeptidation
-
-
-
additional information
?
-
-
reactions catalyzed: 1. peptide bond hydrolysis, 2. ester bond hydrolysis, 3. transesterification, 4. transpeptidation
-
-
-
additional information
?
-
-
reactions catalyzed: 1. peptide bond hydrolysis, 2. ester bond hydrolysis, 3. transesterification, 4. transpeptidation
-
-
-
additional information
?
-
-
alteration of substrate specificity by protein engineering
-
-
-
additional information
?
-
-
little or no reaction with Glu-Phe-p-nitroanilide, benzyloxycarbonyl-Phe-p-nitroanilide, Suc-Phe-p-nitroanilide, acetyl-Phe-p-nitroanilide, Phe-p-nitroanilide, Gly-p-nitroanilide and Ala-p-nitroanilide at 75C, and Val-Leu-Lys-p-nitroanilide, benzoyl-Arg-p-nitroanilide, Suc-Ala-Ala-p-nitroanilide, benzoyl-Tyr-p-nitroanilide, benzyloxycarbonyl-Lys-Arg-p-nitroanilide, and benzyloxycarbonyl-Arg-p-nitroanilide at 40C
-
-
?
additional information
?
-
-
biosynthesis of subtilisin requires participation of an N-terminal prodomain
-
-
?
additional information
?
-
-
does not cleave proPhrE
-
-
-
additional information
?
-
-
autoprocessing of Pro-subtilisin-like serine protease
-
-
-
additional information
?
-
C5VW36
contains the catalytic triad characteristic of subtilisin family proteinases: motif I (Asp200), motif II (His239), and motif III (Ser568). Has the Gram-positive cell wall anchoring motif (Leu-Pro-X-Thr-Gly) at the carboxy-terminus, which is followed by a hydrophobic domain
-
-
-
additional information
?
-
degrades protein components of both nematode (Meloidogyne sp.) and insect (Phthorimaea opercullella) eggs
-
-
-
additional information
?
-
N-succinyl-Ala-Ala-Val-Ala-p-nitroanilide and keratin are poor substrates for AcpII, with relative hydrolysis rates of 1-1.5% of the activity toward N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide or gelatin, respectively
-
-
-
additional information
?
-
-
Sub2 undergoes self-digestion at high concentrations
-
-
-
additional information
?
-
-
substrates bind in a less catalytically favorable conformation after the enzyme has been exposed to organic media for several hours
-
-
-
additional information
?
-
-
displays preference for cleavage after Glu
-
-
-
additional information
?
-
-
enzyme is able to hydrolyze a range of fluorogenic substrates, including AFC derivatives of VEID, VAD, YVAD, IETD, VDVAD and LEHD
-
-
-
additional information
?
-
-
enzymes prefers cleaving after hydrophobic residues (and in particular P1 leucine)
-
-
-
additional information
?
-
-
besides ester hydrolysis the enzyme also performs ester perhydrolysis reacting an ester and a hydroperpxide, residue Gl165 in the S1 binding pocket of the enzyme is involved. Perhydrolysis reaction analysis and kinetcis, overview
-
-
-
additional information
?
-
-
substrate specificity, substrate docking simulations, overview. No activity with Ala-Ala-Pro-Glu-4-nitroanilide and Tyr-Val-Ala-Asp-4-nitroanilide. Substrate specificity and the role of stress signals such as divalent metal ions play roles in defining the proteolytic activity of Bacillus clausii intracellular subtilisin protease, molecular basis, overview. The enzyme unfolds under stress conditions. Heat-denatured whole proteins are found to be better substrates for the enzyme than the native forms
-
-
-
additional information
?
-
the enzyme shows activity towards complex substrates, e.g. skimmed milk
-
-
-
additional information
?
-
-
the enzyme shows activity towards complex substrates, e.g. skimmed milk
-
-
-
additional information
?
-
-
the Fusarium equiseti Fe protease has a broad substrate specificity, almost all amino acid residues are accepted at position P1, even though it shows some preference for cleavage at the C-terminal side of asparagine and histidine residues. The S4 subsite of Fe protease favors aspartic acid and threonine, substrate specificity and comprison to other subtilisin and selected fungal subtilisin-like proteases, overview
-
-
-
additional information
?
-
Bacillus subtilis DB428
-
random mutagenesis to enhance the activity of subtilisin in organic solvents
-
-
-
additional information
?
-
Fusarium equiseti CBS119568
-
the Fusarium equiseti Fe protease has a broad substrate specificity, almost all amino acid residues are accepted at position P1, even though it shows some preference for cleavage at the C-terminal side of asparagine and histidine residues. The S4 subsite of Fe protease favors aspartic acid and threonine, substrate specificity and comprison to other subtilisin and selected fungal subtilisin-like proteases, overview
-
-
-
additional information
?
-
Alkalimonas collagenimarina AC40T
N-succinyl-Ala-Ala-Val-Ala-p-nitroanilide and keratin are poor substrates for AcpII, with relative hydrolysis rates of 1-1.5% of the activity toward N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide or gelatin, respectively
-
-
-
additional information
?
-
C5VW36
contains the catalytic triad characteristic of subtilisin family proteinases: motif I (Asp200), motif II (His239), and motif III (Ser568). Has the Gram-positive cell wall anchoring motif (Leu-Pro-X-Thr-Gly) at the carboxy-terminus, which is followed by a hydrophobic domain
-
-
-
additional information
?
-
Bacillus sp. Ak.1
-
little or no reaction with Glu-Phe-p-nitroanilide, benzyloxycarbonyl-Phe-p-nitroanilide, Suc-Phe-p-nitroanilide, acetyl-Phe-p-nitroanilide, Phe-p-nitroanilide, Gly-p-nitroanilide and Ala-p-nitroanilide at 75C, and Val-Leu-Lys-p-nitroanilide, benzoyl-Arg-p-nitroanilide, Suc-Ala-Ala-p-nitroanilide, benzoyl-Tyr-p-nitroanilide, benzyloxycarbonyl-Lys-Arg-p-nitroanilide, and benzyloxycarbonyl-Arg-p-nitroanilide at 40C
-
-
?
additional information
?
-
Bacillus subtilis 72
-
influence of substrate structure of N-protected peptide nitroanilides of the types: benzyloxycarbonyl-A2-A1 4-nitroanilide, benzyloxycarbonyl-A3-A2-A1 4-nitroanilide, benzyloxycarbonyl-A4-A3-A2-A1 4-nitroanilide, subsite S1 is of broad selectivity: preference for hydrophobic amino acid residues (i.e. leucine and phenylalanine) the beta-branched and the basic amino acid residues cannot interact with the S1 subsite and the hydrolysis of the corresponding peptides occurs exclusively at the A2-A1 bond. If S1/A1 interactions are weak (Ala, norvaline, norleucine) the amino acid residue A1 can interact with subsites S1 and S1' resulting in the hydrolysis of two bonds (A1 4-nitroanilide and A2-A1). The subsite S2 reveals a preference for small amino acid residues
-
-
-
additional information
?
-
Bacillus sp. GX6644
-
high elastolytic acitivity, preferentially hydrolyzes the ester bond of Ala, but significant hydrolysis is observed with other aliphatic (Gly, Leu) and aromatic (Tyr) amino acids
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
biosynthesis of subtilisin requires participation of an N-terminal prodomain
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
no requirement for a cofactor
-
additional information
-
does not depend on a cofactor
-
additional information
does not depend on a cofactor
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
-
as found in other Bacillus subtilisins, the structure of wild-type Savinase contains 2 calcium ion-binding sites
Ca2+
-
stabilizing mutations are usually calcium-dependent in their stabilizing effect
Ca2+
-
bound calcium ions play a key role in protecting against autolysis and thermal denaturation
Ca2+
-
increase on activity by 20%
Ca2+
-
two binding sites, important for correct folding
Ca2+
-
three binding sites, important for correct folding
Ca2+
-
five binding sites, important for correct folding
Ca2+
three binding sites, important for correct folding
Ca2+
-
six calcium ions bind to pro-S255A at the loop regions. The at least six Ca2+ ions bind to pro-S255A too tightly to be removed by dialysis
Ca2+
-
in the presence of calcium the half-life at 55C and 60C are 3.6fold and 3.48fold higher for the native enzyme compared to that in the absence of added calcium. In the presence of 10 mM calcium the half-life of the enzyme at 60C increases by 6.06fold, 5.20fold and 2.92fold when coupled with oxidized sucrose polymers OSP400, OSP70 and polyglutaraldehyde, respectively
Ca2+
-
calcium ion binds weakly to the Ca-7 site in the unautoprocessed form, but is trapped upon autoprocessing. The Ca-7 site is required to promote the autoprocessing reaction by stabilizing the autoprocessed form, in which the new N-terminus of the mature domain is structurally disordered
Ca2+
is required for catalysis; required for catalysis; required for catalysis; required for catalysis
Ca2+
enzyme activity and/or stability depends on the presence of divalent cations, probably Ca2+ ions, near the surface of the enzyme
Ca2+
-
Ca2+-binding loop is required for folding of subtilisin but does not seriously contribute to the stabilization of subtilisin in a native structure
Ca2+
calcium-loaded state of five ions bound to each of the two subtilisin molecules. Three of the binding sites have two side chains of an acidic residue coordinating the calcium ion, whereas the other two binding sites have either a main-chain carbonyl, or only one acidic residue side chain coordinating the calcium ion; calcium-loaded state of five ions bound to each of the two subtilisin molecules. Three of the binding sites have two side chains of an acidic residue coordinating the calcium ion, whereas the other two binding sites have either a main-chain carbonyl, or only one acidic residue side chain coordinating the calcium ion
Ca2+
the enzyme contains 7 Ca2+ ions, 4 of which are responsible for folding requirement of Ca2+ ions for the hyperthermostability of Tk-subtilisin, the Ca1, Ca6, and Ca7 ions, especially the Ca1 ion, contribute to the hyperthermostabilization of Tk-subtilisin
Ca2+
-
required, the presence of a low-affinity Ca2+ binding site formed by the backbone carbonyls cannot be excluded based on the amino acid sequence comparisons
Ca2+
-
required for thermostability
Ca2+
-
is important for ISP activity through structural changes. The looped turn constituted by residues Ala180-Pro197 binds Ca2+. Removal of Ca2+ at sites close to the dimer interface and the S1 pocket are involved enzyme inhhibition by EDTA
Cd2+
-
stabilizes
Cl-
treatment with CsCl increases the activity several fold. Two Cl- ions are close to the mouth of the active site cleft, where they may affect catalysis
Cs+
treatment with CsCl increases the activity several fold. Two Cs+ ions are close to the mouth of the active site cleft, where they may affect catalysis
Cu2+
-
5 mM, 40% increase in activity
Cu2+
-
100% inhibition
KCl
increases enzymatic activity somewhat, while choline-Cl has a larger effect
Mg2+
-
100% inhibition
Sr2+
-
increases thermostability, but to a significantly lower degree than Ca2+
Mg2+
-
required
additional information
-
enzyme contains no metal ion
additional information
-
shows no evidence of Ca2+ activation
additional information
-
strain DJ-4, not significantly affected by Ca2+, Co2+, Mg2+, or Mn2+
additional information
-
metal ion sites are not formed until late in folding and do not influence the kinetics of folding to the intermediate complex
additional information
enzymatic activity of subtilisin crystals in acetonitrile is sensitive to the type of counterions present before transfer to the organic solvent. Larger cations increase the enzyme activity
additional information
-
unlike subtilisin, subtilisin-like serine protease does not require Ca2+ for folding
additional information
-
metal ions are not involved in the subtilisin catalytic mechanism but are an important structural element
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(dimethylamino)-1-naphthalenesulfonyl fluoride
-
100% inhibition of enzyme activity with N-trans cinnamoyl imidazole as substrate, during prolonged exposure to organic solvents the active-site fluorescent label inhibitor adopts a different binding conformation
1,10-phenanthroline
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 40%
1,4-dioxane
-
104fold decrease in the enzymes catalytic activity for the hydrolysis reaction of vinyl butyrate with D2O and a 50% decrease in enzyme structural dynamics. Attaching increasing amounts of PEG to the enzyme reverses some of the activity loss. Poly(ethylene)-glycolylation increases protein structural dynamics in 1,4-dioxane
1,4-dioxane
-
reduces the enzyme activity. Structural changes, flexibility, hydration, and changes of the enzyme ionization state are not responsible for the low storage stability. Possible depletion or rearrangement of water molecules around the active site, or small structural perturbations around the active site or movements of counter ions
2,2'-[[(1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphoryl]bis(oxybenzene-4,1-diyl)]diacetic acid
-
-
2,2'-[[(1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphoryl]bis(oxybenzene-4,1-diyl)]diacetic acid
-
-
2,2'-[[(1-[[1-(tert-butoxycarbonyl)-L-prolyl]amino]-3-methylbutyl)phosphoryl]bis(oxybenzene-3,1-diyl)]diacetic acid
-
-
2-phenylethaneboronic acid
-
-
2-phenylethanesulfonic acid
-
-
3,4-dichloroisocoumarin
-
-
4-(2-aminoethyl)-benzenesulfonyl fluoride
-
-
4-(2-aminoethyl)benzenesulfonyl fluoride
-
-
4-(4'-Aminophenylazo)phenylarsonic acid
-
-
4-bromophenacyl bromide
-
-
5-Dimethylaminonaphthalene-1-sulfonate
-
circular-dichroism and fluorescence studies of 5-dimethylaminonaphthalene-1-sulfonyl derivative of subtilisin indicate a closely similar structure to that of native subtilisin
acetone
-
-
acetonitrile
-
-
alpha2-Macroglobulin
-
inhibits alkaline proteolytic activity of purified Arp by 80%
-
antipain
-
-
antipain
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 75%
antipain
-
totally inhibits recombinant Sub2
Aprotinin
-
inhibits the enzymatic activity by 15%
Benzamide
-
-
benzeneboronic acid
-
-
Benzenesulfonic acid
-
-
Benzyloxycarbonyl-(Ala)n-PheCH2Cl
-
benzyloxycarbonyl-(Ala)2-PheCH2Cl is the best inhibitor
Benzyloxycarbonyl-Ala-XaaCH2Cl
-
Xaa : Gly, Val, Ala, Leu, Phe
Benzyloxycarbonyl-L-phenylalanylbromomethane
-
reactivity is about an order of magnitude less than that of subtilisins BPN' and Carlsberg
bis(2,3,5-trimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(2,3-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(2,3-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(2,5-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(2-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(2-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(3,4,5-trimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(3,4,5-trimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(3,4-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(3,4-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(3-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(3-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(3-methoxyphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(4-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(4-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(4-ethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(4-ethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(4-methoxyphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(4-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis(4-tert-butylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis(4-tert-butylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis[4-(methylsulfonyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis[4-(propan-2-yl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis[4-(propan-2-yl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
bis[4-(sulfanylmethyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
bis[4-(sulfanylmethyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
Brij 35
at 37C and pH of 7.5, 0.01% reduces prosubtilisin JB1 relative activity to 30% and 0.05% reduces prosubtilisin JB1 relative activity to 70%
Broad bean extract
-
subtilisin BPN'
Ca2+
at 37C and pH of 7.5, 1 mM reduces prosubtilisin JB1 relative activity to 57% and 5 mM reduces prosubtilisin JB1 relative activity to 45%
carbon
-
absence of C or N metabolite repression in subtilisins from KV42 isolate. Together with carbon significantly reduces protease activity of KV71 at 16 and 36 h, and KV22 at 16 h
-
carbon
-
together with carbon significantly reduces protease activity of KV01 at 16 and 36 h, and KV54 at 16 h
-
Chiral amine- and aminoalcohol-derivatives
-
-
-
chymostatin
-
-
chymostatin
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 95%
chymostatin
-
totally inhibits recombinant Sub2
Chymotrypsin I inhibitor from potato
-
subtilisin Carlsberg, subtilisin BPN'
-
chymotrypsin inhibitor 2
-
-
-
chymotrypsin inhibitor 2 mutant M59A
-
-
-
chymotrypsin inhibitor 2 mutant M59F
-
-
-
chymotrypsin inhibitor 2 mutant M59G
-
-
-
chymotrypsin inhibitor 2 mutant M59K
-
-
-
chymotrypsin inhibitor 2 mutant M59Y
-
-
-
chymotrypsin inhibitor 2 mutant Y61A
-
-
-
Co2+
at 37C and pH of 7.5, 1 mM reduces prosubtilisin JB1 relative activity to 14% and 5 mM reduces prosubtilisin JB1 relative activity to 24%
CrSPI-1
synthetic construct
-
Kazal-type inhibitor from the hepatopancreas of the Carcinoscorpius rotundicauda, potently inhibits subtilisin
-
Cu2+
-
strain DJ-4, strong inhibition
Cu2+
at 37C and pH of 7.5, with 1 mM results in an almost complete reduction of prosubtilisin JB1 activity, 5 mM reduces prosubtilisin JB1 relative activity to 19%
Cyclohexanol
-
-
cyclohexanone
-
-
diisopropyl fluorophosphate
-
-
diisopropylphosphofluoridate
-
-
Dipeptidyl chloromethyl ketones
-
-
diphenyl (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
-
diphenyl (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
-
duck ovomucoid
-
-
-
E-64
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 31%
E-64c
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 25%
EDTA
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 52%
EDTA
5 mM partially inhibits by 35%, in 100 mM Tris-HCl buffer, pH 7.0 at 15C for 20 min
EDTA
-
subtilisin exhibits little activity at 80C in the presence of 10 mM. Subtilisin-like serine protease is fully active even in the presence of 10 mM EDTA
EDTA
-
inactivates the enzyme at 0.01 mM, the enzyme protein becomes less structured and potentially monomeric. Removal of Ca2+ at sites close to the dimer interface and the S1 pocket are involved enzyme inhhibition by EDTA
Eglin c
-
-
EGTA
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 34%
Elastatinol
-
weak
EPI1a
-
four-cysteine atypical Kazal-domain of protease inhibitor EPI1 from Phytophthora infestans. 80% inhibition at 0.00015 mM
-
EPI1b
-
typical Kazal-domain of protease inhibitor EPI1 from Phytophthora infestans, little inhibitory effect
-
fungal protease inhibitor F
-
specific inhibitor toward subtilisin-type protease. P1 residue most signficantly affects inhibitory specificity. Mutant T29M has stronger subtilisin-inhibitory activity than the wild-type, mutants T29E and T29R are relatively weaker inhibitors. Inhibitory activities of mutants T29F and T29L are as strong as that of the wild-type
-
guanidine hydrochloride
-
at 2 M inhibits subtilisin-like serine protease by 35% and at 4 M almost completely, whereas it has no inhibitory effect on subtilisin
guanidinium hydrochloride
-
-
Hg+
at 37C and pH of 7.5, 1 mM reduces prosubtilisin JB1 relative activity to 33% and 5 mM reduces prosubtilisin JB1 relative activity to 22%
human LEKTI
-
noncompetitive inhibition
-
human proteinase inhibitor 9
-
PI9, serine proteinase inhibitor
-
Hydrocinnamamide
-
-
Hydrocinnamate
-
-
indole
-
-
Inhibitor from Dolichos biflorus
-
purification and properties
-
Inhibitor from egg white
-
subtilisin Carlsberg, subtilisin BPN'
-
Inhibitor from egg white
-
-
-
Inhibitor from seeds of Canavalia lineata
-
low MW protein of about 6500 MW
-
Inhibitor from seeds of Canavalia lineata
-
MW 22000, Kunitz type inhibitors
-
Inhibitor from seeds of Setaria italica
-
purification and characterization
-
Inhibitor from Streptomyces sp.
-
-
-
Inhibitor from Streptomyces virginiae
-
primary structure
-
Inhibitor from Vigna unguiculata subsp. cylindrica
-
inactive with other enzymes
-
Inhibitor from Vigna unguiculata subsp. cylindrica
-
properties and kinetics of the inhibitor
-
Inhibitor of Amaranthus caudatus seeds
-
subtilisin Novo (BPN') from Bacillus amyloliquefaciens, subtilisin Carlsberg from Bacillus licheniformis
-
Inhibitor of trypsin from soybean
-
-
-
iodoacetamide
-
-
K+
at 37C and pH of 7.5, with 1 mM results in an almost complete reduction of prosubtilisin JB1 activity, 5 mM reduces prosubtilisin JB1 relative activity to 53%
L-[(1R)-1-acetamido-2-(1-naphthyl)ethyl]boronic acid
-
-
Leupeptin
-
inhibits the enzymatic activity by 20%
Methaneboronic acid
-
-
N-((tert-Butoxycarbonyl)alanylprolylphenylalanyl)-O-benzoylhydroxylamine
-
-
N-(tert-butoxycarbonyl)-L-valyl-N-(1-[bis[4-(sulfanylmethyl)phenoxy]phosphoryl]-2-phenylethyl)-L-prolinamide
-
-
N-(tert-butoxycarbonyl)-L-valyl-N-[1-(diphenoxyphosphoryl)-2-phenylethyl]-L-prolinamide
-
-
N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(3,4,5-trimethylphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
-
-
N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(4-methoxyphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
-
-
N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(4-tert-butylphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
-
-
N-acetyl-L-tryptophan amide
-
molecular dynamics simulations are performed with subtilisin in the presence and in the absence of an inhibitor both in hexane and in water. The inhibitor induces an open conformation of the S1 pocket that is maintained after the removal of the ligand in anhydrous, but not in aqueous, simulations. The analysis of fluctuations suggest that this behavior is caused by the decreased flexibility exhibited by subtilisin in hexane
N-benzoyl-L-Arg
-
product inhibition
N-benzyloxycarbonyl-Ala-Pro-Phe-chloromethyl ketone
-
synthetic inhibitor
N-ethylmaleimide
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 15%
N-tosyl-L-phenylalanyl chloromethyl ketone
-
-
N1-18[ISP]
-
synthetic peptide corresponding to the N-terminal extension behaves as a mixed noncompetitive inhibitor of active ISP
nitrogen
-
absence of C or N metabolite repression in subtilisins from KV42 isolate. Together with carbon significantly reduces protease activity of KV71 at 16 and 36 h, and KV22 at 16 h
nitrogen
-
together with carbon significantly reduces protease activity of KV01 at 16 and 36 h, and KV54 at 16 h
p-Arsanilate
-
-
p-chloromercuribenzoate
-
partial inhibition
p-Nitrophenylarsonate
-
-
p-Tolylarsonate
-
-
Pepstatin
-
almost completely inhibits acid proteolytic activity, does not inhibit alkaline proteolytic activity of purified Arp
pepstatin A
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 48%
Peptidyl chloromethyl ketones
-
-
Phenol
-
-
Phenylarsonate
-
-
phenylmethanesulfonyl fluoride
-
-
phenylmethanesulfonyl fluoride
-
-
phenylmethanesulfonyl fluoride
-
totally inhibits recombinant Sub2
phenylmethylsulfonyl fluoride
-
strain DJ-4
phenylmethylsulfonyl fluoride
-
-
phenylmethylsulfonyl fluoride
-
1 mM, 82% inhibition
phenylmethylsulfonyl fluoride
-
-
phenylmethylsulfonyl fluoride
-
1 mM, 5% residual activity
phenylmethylsulfonyl fluoride
-
inhibits alkaline proteolytic activity of purified Arp by 100%, does not inhibit acid proteolytic activity
phenylmethylsulfonyl fluoride
at 37C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 79%
phenylmethylsulfonyl fluoride
1 mM completely inhibits, in 100 mM Tris-HCl buffer, pH 7.0 at 15C for 20 min
Phenylmethylsulphonyl fluoride
-
-
PMSF
-
circular-dichroism and fluorescence studies of PMSF derivative of subtilisin indicate a closely similar structure to that of native subtilisin
Potato extract
-
subtilisin BPN'
-
propeptide
-
inhibition in a concentration-dependent manner. Wild-type propeptide is more potent than G56W-, G56S- and G56E-propeptide
-
RNA aptamer
-
-
-
Sodium dodecyl sulfate
-
activity of the enzyme is retarded by 2.3 and 244times in 1 mM and 40 mM sodium dodecyl sulfate respectively compared to that in buffer solution. No evidence of sandwich-like subtilisinsodium dodecyl sulfate complex formation, thus the enzyme does not encroach into the hydrophobic surfactant core of sodium dodecyl sulfate micelle to form an elongated structure. Retains its structural integrity in sodium dodecyl sulfate solution. Micellar crowding in the vicinity of the enzyme
Sodium dodecyl sulfate
at 37C and pH of 7.5, 0.01% reduces prosubtilisin JB1 relative activity to 52% and 0.05% reduces prosubtilisin JB1 relative activity to 29%
Suc-Val-Pro-PheP(OPh)2
-
-
tert-butyl (2S)-2-([1-[bis(2-methylphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
-
tert-butyl (2S)-2-([1-[bis(3,4,5-trimethylphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
-
tert-butyl (2S)-2-([1-[bis(3,4-dimethylphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
-
tert-butyl (2S)-2-([1-[bis(4-methoxyphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
-
tert-butyl (2S)-2-([1-[bis(4-methylphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
-
tert-butyl (2S)-2-[(1-[bis[4-(sulfanylmethyl)phenoxy]phosphoryl]-2-phenylethyl)carbamoyl]pyrrolidine-1-carboxylate
-
-
tert-butyl (2S)-2-[[1-(diphenoxyphosphoryl)-2-phenylethyl]carbamoyl]pyrrolidine-1-carboxylate
-
-
tomato inhibitor-II
-
TI-II, enzyme binding structure, the interdomain interface in TI-II consists of a small cluster of highly conserved hydrophobic residues Ile14, Pro16, Tyr98, Phe100 and Phe106 from domain I and Tyr34, Pro54 and Lys55 from domain II. Although this interface is quite small (buried surface area of 487A), it forms a stable packing arrangement between the two domains. Each reactive site loop in TI-II interacts with a separate molecule of subtilisin in the canonical manner observed in other proteinaseinhibitor complexes. The domains of TI-II appear to bind the proteinase independently of each other
-
tosyl-Phe chloromethyl ketone
-
not
tosyl-Phe chloromethyl ketone
-
-
Triton X-100
at 37C and pH of 7.5, 0.01% reduces prosubtilisin JB1 relative activity to 92% and 0.05% reduces prosubtilisin JB1 relative activity to 69%
turkey egg white inhibitor
-
-
-
Tween 20
at 37C and pH of 7.5, 0.01% reduces prosubtilisin JB1 relative activity to 53% and 0.05% reduces prosubtilisin JB1 relative activity to 70%
Zn2+
-
strain DJ-4, strong inhibition
Zn2+
-
1 mM, 59% residual activity
Zn2+
at 37C and pH of 7.5, 1 mM reduces prosubtilisin JB1 relative activity to 30%, with 5 mM results in an almost complete reduction of prosubtilisin JB1 activity
Mg2+
at 37C and pH of 7.5, 1 mM reduces prosubtilisin JB1 relative activity to 79% and 5 mM reduces prosubtilisin JB1 relative activity to 48%
additional information
-
not: tosyl-leucine chloromethyl ketone
-
additional information
-
metal chelating reagents; not: tosyl-leucine chloromethyl ketone; sulfhydryl reagents
-
additional information
-
strain DJ-4, no inhibition by EDTA or leupeptin
-
additional information
-
not inhibitory: Tween-20, Tween-40, Tween-60, Tween-80 at 1%, sodium dodecylsulfate at 0.2%, for 1 h at 30C. Not inhibitory: iodoacetate, ethylacetimide, phenylglyoxal, N-ethylmaleimide, N-bromosuccinimide
-
additional information
-
not inhibitory: EDTA at 5 mM
-
additional information
-
enzyme-specific, thermostable inhibitor from Physarum polycephalum with molecular mass of 32-33 kDa, 50% inhibition at 0.00014 mM
-
additional information
-
not inhibitory: sodium dodecylsulfate at 1 mM
-
additional information
-
not inhibited by 1,10 phenanthroline
-
additional information
-
alkaline proteolytic activity of purified Arp not inhibited by 3,4-dichloroisocumarin, tosyl lysine chloromethyl ketone, tosyl phenylalanine chloromethyl ketone, leupeptin, chymostatin, elastatinal, iodoacetamide, E-64, EDTA, 1,10-phenanthroline, aprotinin, antithrombin III, alpha1-antitrypsin and soybean trypsin inhibitor
-
additional information
-
enzymatic activity of mature subtilisin is inhibited by G56S-propeptide and wild-type propeptide in a concentration-dependent manner
-
additional information
at 37C and pH of 7.5, 0.1 mM leupeptin does not inhibit prosubtilisin JB1
-
additional information
-
is not inhibited by 1,10-phenanthroline, EDTA, E64, bestatin, phosphoramidon and pepstatin A
-
additional information
-
subtilisin-like serine protease is fully resistant to treatment with 2-5% SDS, 4-8 M urea, 10% Tween-20 or 10% Triton X-100. In gel assay, subtilisin-like serine protease is fully denatured prior to SDS-PAGE by trichloroacetic acid treatment, followed by boiling for 5 min in the presence of SDS
-
additional information
-
aldehyde and fluoromethyketone (FMK)-based inhibitors inhibit SAS-1 and -2 reversibly and only CMK-based inhibitors irreversibly inhibit the enzyme
-
additional information
-
molecular dynamics simulations, residue displacement correlations, and inhibitor design based on the enzyme inhibitor comlpex of TI-II and subtilisin, overview
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
choline
increases the enzyme activity
dichloroisocoumarin
-
-
n-Propanol
-
subtilisin precipitated and rinsed with propanol yields higher transesterfication activity. About 10000times increase in initial rates in 1-butyl-3-methylimidazolium hexafluorophosphate over what is obtained with pH tuned lyophilized powders
OSP400
-
in the presence of 10 mM calcium the half-life of the enzyme at 60C increases by 6.06fold. Among modifiers used OSP400 is most effective in stabilizing the enzyme. Thermally induced unfolding is delayed
-
OSP70
-
in the presence of 10 mM calcium the half-life of the enzyme at 60C increases by 5.20fold. Thermally induced unfolding is delayed
-
poly(ethylene glycol)
-
poly(ethylene glycol)-modified subtilisin suspended in ionic liquids exhibits excellent catalytic performance while the native enzyme shows no activity
poly(ethyleneglycol)
-
presence of poly(ethyleneglycol) combined with trehalose during lyphilization does help in obtaining a more active enzyme preparation for catalysis in ionic liquids
polyglutaraldehyde
-
in the presence of 10 mM calcium the half-life of the enzyme at 60C increases by 2.92fold. Thermally induced unfolding is delayed
Toluene
-
poly(ethylene glycol)-modified subtilisin
[C2OC1mim][Tf2N]
-
poly(ethylene glycol)-modified subtilisin
-
[C2OHmim][Tf2N]
-
poly(ethylene glycol)-modified subtilisin
-
[Emim][Tf2N]
-
poly(ethylene glycol)-modified subtilisin shows good stability in [Emim][Tf2N], and maintains 80% of its initial activity after 60 h. Preferred medium for enzymatic reaction. Enzyme activity is much higher than in conventional organic solvents. Excellent activity is associated with unique properties such as hydrophobicity and high polarity
-
methyl-beta-cyclodextrin
-
activates and enhances the enantioselectivity of subtilisin C. mainly by prevention of structural perturbation during the initial lyophilization process. Minor contributions to increased activity likely stem from other factors, such as, reduction of possible mass transport limitations and changes in enzyme dynamics. Enhanced activity and enantioselectivity is not caused by formation of cyclodextrinsubstrate or -product complexes
additional information
-
conjugation of enzyme with comb-shaped poly-(ethylen glycol) and solubilization in ionic liquids without adding water. Enzyme exhibits higher transesterification activity in solution of [Eminm][Tf2N] than in toluene. No enzymic activity in DMSO, THF, or acetonitrile
-
additional information
-
lyophilized enzyme, sharp increase in enzyme activity in correlation with an increasing amount of fumed silica added to enzyme before freeze-drying. When freezing at -20C instead of liquid nitrogen, even better activation is observed
-
additional information
-
prodomain mutant proR9 accelerates subtilisin folding. Subtilisin folding with the wild-type prodomain is slow. The wild-type prodomain binds 100times slower than proR9
-
additional information
-
three phase partitioning reveals 13fold increase in the initial transesterfication rate
-
additional information
-
improvement in the enzymatic activity for the poly(ethylene)-glycol-modified enzyme in the organic solvent that depends on the level of poly(ethylene)-glycolylation. PEGSBc conjugates with 1.1, 1.9, and 3.2 average mol of poly(ethylene)-glycol per mol of protein reveal a 30fold, 57fold, and 99fold increase in enzymatic activity as a result of poly(ethylene)-glycolylation
-
additional information
-
sigma-H is required for full expression of the CSF-processing protease
-
additional information
-
no activity of poly(ethylene glycol)-modified subtilisin in methyl tert-butyl ether, tetrahydrofuran, dimethylsulfoxide, and acetonitrile
-
additional information
-
dynamics are also affected by the method of preparation of the enzyme: decreased flexibility is observed when the enzyme is chemically modified with poly ethylene glycol or colyophilized with crown ethers
-
additional information
synthetic construct
-
glycosylation of subtilisin does not cause any significant tertiary structure changes
-
additional information
-
unlike subtilisin, subtilisin-like serine protease does not require propeptide for folding
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.85
acetyl-L-Phe
-
30C, pH 8.0
0.22
acetyl-L-Phe ethyl ester
-
30C, pH 8.0
4
acetyl-L-Tyr
-
30C, pH 8.0
-
0.37
acetyl-L-Tyr ethyl ester
-
30C, pH 8.0
7
benzoyl-Arg ethyl ester
-
subtilisin Novo, subtilisin Carlsberg
10
benzoyl-Arg ethyl ester
-
subtilisn BPN'
2.1
benzoyl-L-Arg ethyl ester
-
30C, pH 8.0
8.3
benzyloxycarbonyl-Gly-p-nitroanilide
-
pH 7.5, 20C, in presence and absence of DTT
0.0054
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-4-nitroanilide
-
-
0.059
casein
-
pH 10, 45C, Vmax: 0.336 mg/min
0.13
Glp-Ala-Ala-Leu-p-nitroanilide
-
in 2% DMSO
0.18
Glp-Ala-Ala-Leu-p-nitroanilide
-
in 2% DMF
0.25
Glp-Ala-Ala-Leu-p-nitroanilide
-
in 2% DMF
0.98
L-Ala-L-Ala-L-Pro-L-Ala
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
1.99
L-Ala-L-Ala-L-Pro-L-Ala
-
wild-type, pH 9.0, 24C
1.15
L-Ala-L-Ala-L-Pro-L-Phe
-
wild-type, pH 9.0, 24C
1.93
L-Ala-L-Ala-L-Pro-L-Phe
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
0.039
L-Ala-L-Ala-L-Val-L-Ala
-
wild-type, pH 9.0, 24C
0.053
L-Ala-L-Ala-L-Val-L-Ala
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
0.223
L-Phe-L-Ala-L-Ala-L-Phe
-
wild-type, pH 9.0, 24C
0.24
L-Phe-L-Ala-L-Ala-L-Phe
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
0.62
L-Tyr-L-Val-L-Ala-L-Asp
-
wild-type, pH 9.0, 24C
0.75
L-Tyr-L-Val-L-Ala-L-Asp
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
0.15
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide
-
subtilisin DY
0.15
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide, subtilisin BPN'
0.15
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide
-
-
120
N-Acetyl-Ala methyl ester
-
subtilisin BPN'
66
N-Acetyl-Leu methyl ester
-
subtilisin BPN'
91
N-Acetyl-Lys methyl ester
-
subtilisin BPN'
17
N-acetyl-Phe ethyl ester
-
subtilisin BPN'
60
N-Acetyl-Phe methyl ester
-
subtilisin Novo
24
N-Acetyl-Trp ethyl ester
-
subtilisin BPN'
50
N-Acetyl-Trp methyl ester
-
subtilisin Carlsberg
22
N-Acetyl-Tyr ethyl ester
-
subtilisin BPN'
70
N-Acetyl-Tyr ethyl ester
-
subtilisin Novo, N-acetyl-Tyr methyl ester, subtilisin Carlsberg
90
N-Acetyl-Tyr methyl ester
-
subtilisin Novo, N-acetyl-Tyr ethyl ester, subtilisin Carlsberg, N-acetyl-Trp methyl ester, subtilisin Novo
3.13
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
-
pH 8.5, 37C
3.74
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
-
pH 8.5, 37C, mutant D182G
3.84
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
-
pH 8.5, 37C, wild-type
4.02
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
-
pH 8.5, 37C, mutant D180G/D182G
4.28
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
-
pH 8.5, 37C, mutant D180G
0.15
N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 7.4, 37C, mutant S101W/G169A/V192A
0.35
N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 7.4, 37C, wild-type
0.15
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
recombinant mutant T58A/L216W, pH 7.5, 30C
0.2
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
recombinant mutant T58A/G165Y/L216W, pH 7.5, 30C
0.227
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 8.0, 25C, recombinant mutant N212G
0.233
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 8.0, 25C, recombinant wild-type enzyme
0.278
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 8.0, 25C, recombinant mutant P210G
0.279
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 8.0, 25C, recombinant wmutant T211G
0.29
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 8.0, 25C, recombinant mutant P210G/T211G
0.301
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 8.0, 25C, recombinant mutant P210G/T211G/N212G
0.325
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
pH 8.0, 25C, recombinant mutant P210A
0.42
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
recombinant mutant T58A/G165L/L216W, pH 7.5, 30C
0.54
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
recombinant mutant T58A/G165I/L216W, pH 7.5, 30C
0.59
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
-
recombinant wild-type enzyme, pH 7.5, 30C
0.11
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
-
subtilisin-like serine protease, at 20C, in 50 mM Tris-HCl buffer, pH 7.5
0.41
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
-
subtilisin-like serine protease, at 80C, in 50 mM Tris-HCl buffer, pH 7.5
2.4
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
-
subtilisin, at 20C, in 50 mM Tris-HCl buffer, pH 8.0
7.9
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
-
subtilisin, at 80C, in 50 mM Tris-HCl buffer, pH 8.0
0.000655
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide
-
37C, pH 10.5
130
sec-phenethyl alcohol
-
after incubation in 1,4-dioxane, co-lyophilization with methyl-beta-cyclodextrin
319
sec-phenethyl alcohol
-
after a 4-day incubation period in 1,4-dioxane, co-lyophilization with methyl-beta-cyclodextrin
0.28
Suc-Ala-Ala-Ala-p-nitroanilide
-
pH 7.5, 5C
0.32
Suc-Ala-Ala-Ala-p-nitroanilide
-
pH 7.5, 20C
0.58
Suc-Ala-Ala-Ala-p-nitroanilide
-
pH 7.5, 20C, DTT
0.63
Suc-Ala-Ala-Ala-p-nitroanilide
-
pH 7.5, 40C
2.25
Suc-Ala-Ala-Ala-p-nitroanilide
-
pH 7.5, 75C
4
Suc-Ala-Ala-p-nitroanilide
-
pH 7.5, 20C
6.67
Suc-Ala-Ala-p-nitroanilide
-
pH 7.5, 20C, DTT
17
Suc-Ala-Ala-p-nitroanilide
-
pH 7.5, 40C
0.33
Suc-Ala-Ala-Pro-Ala-p-nitroanilide
-
pH 7.5, 5C or 40C
0.92
Suc-Ala-Ala-Pro-Ala-p-nitroanilide
-
pH 7.5, 75C
0.68
Suc-Ala-Ala-Pro-Leu-p-nitroanilide
-
pH 7.5, 5C
1.01
Suc-Ala-Ala-Pro-Leu-p-nitroanilide
-
pH 7.5, 40C
3.37
Suc-Ala-Ala-Pro-Leu-p-nitroanilide
-
pH 7.5, 75C
0.0027
Suc-Ala-Ala-Pro-Phe-4-nitroanilide
synthetic construct
-
1 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25C
0.0036
Suc-Ala-Ala-Pro-Phe-4-nitroanilide
synthetic construct
-
2.2 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25C
0.0037
Suc-Ala-Ala-Pro-Phe-4-nitroanilide
synthetic construct
-
3.5 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25C; subtilisin without bound lactose, in 10 mM potassium phosphate buffer, pH 7.8, at 25C
0.07
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
-
pH 7.5, 5C
0.15
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
-
pH 7.5, 20C
0.3
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
-
pH 7.5, 40C
0.44
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
-
pH 7.5, 20C, DTT
1
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
-
pH 7.5, 75C
0.8
succinyl-AAPA-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-c
1.74
succinyl-AAPA-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-g
1.9
succinyl-AAPA-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-e and S166S-f
2
succinyl-AAPA-p-nitroanilide
-
pH 8.6, 25C wild-type
1.9
succinyl-AAPE-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-a
4.4
succinyl-AAPE-p-nitroanilide
-
pH 8.6, 25C wild-type
0.39
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S166C-S-a
0.51
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S166C
0.54
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S156C-S-d
0.55
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C, wild-type and mutant M222C-S-e
0.6
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S156C-S-c and S166C-S-f
0.61
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant M222C-S-f
0.63
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant M222C-S-b
0.65
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S156C
0.67
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant M222C-S-d
0.68
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S156C-S-b and M222-S-a
0.68
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C, mutant S166C-a
0.69
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant M222C-S-c
0.7
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S166C-S-e
0.7
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C, mutant S166C-f
0.72
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S166C-S-c
0.73
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C wild-type
0.74
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S166C-S-d
0.74
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C, mutant S166C-g
0.77
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant M222C
0.78
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S156C-S-e
0.79
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S156C-S-a
0.86
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S156C-S-f
0.92
succinyl-AAPF-p-nitroanilide
-
pH 7.5, 25C mutant S166C-S-b
1.09
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-e
1.17
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-c
1.34
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-b
1.5 - 2
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-h
1.6
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-d
2.26
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-i
2.46
succinyl-AAPF-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-j
3.4
succinyl-AAPR-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-b
5.2
succinyl-AAPR-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-j
5.3
succinyl-AAPR-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-i
5.5
succinyl-AAPR-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-d
7.2
succinyl-AAPR-p-nitroanilide
-
pH 8.6, 25C wild-type
8.2
succinyl-AAPR-p-nitroanilide
-
pH 8.6, 25C mutant S166C-S-h
1.2
Succinyl-Ala-Ala-Ala 4-nitroanilide
-
subtilisin Carlsberg
1.5
Succinyl-Ala-Ala-Ala 4-nitroanilide
-
subtilisin DY
0.71
succinyl-Ala-Ala-Phe 4-nitroanilide
-
subtilisin DY
0.79
succinyl-Ala-Ala-Phe 4-nitroanilide
-
subtilisin Carlsberg
0.13
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide
-
mutant subtilisin BPN' Asp to Ser99
0.13
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide
-
-
0.13
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide
-
succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl 7-amide, subtilisin Sendai
0.36
succinyl-L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
-
mutant Y217L, pH 8.3, 25C
0.45
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide
-
mutant Y217L, pH 8.3, 25C
0.33
succinyl-L-Asp-L-Val-L-Arg-L-Ala-L-Phe-7-amido-4-methylcoumarin
-
mutant Y217L, pH 8.3, 25C
30
toluenesulfonyl-Arg methyl ester
-
subtilisin Novo, N-acetyl-Phe methyl ester, subtilisin Carlsberg
40
toluenesulfonyl-Arg methyl ester
-
subtilisin Carlsberg
0.09
Urea-denatured hemoglobin
-
30C, pH 8.0
-
0.14
Z-Ala-Ala-Leu-p-nitroanilide
-
in 20% DMSO
0.19
Z-Ala-Ala-Leu-p-nitroanilide
-
in 20% DMF
0.25
Z-Ala-Ala-Leu-p-nitroanilide
-
in 20% DMSO
4
Z-Ala-Ala-Leu-p-nitroanilide
-
in 20% DMF
0.22
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide
-
subtilisin Carlsberg
additional information
additional information
-
-
-
additional information
additional information
-
-
-
additional information
additional information
-
the temperature dependence of the kinetics and thermodynamic parameters suggest that the enzyme exists in two, i.e. cold and hot forms, at 22C the cold form turns into the hot one possibly owing to a conformational change
-
additional information
additional information
-
-
-
additional information
additional information
-
Km of subtilisin Novo chemically attached to soluble DEAE-dextran and insoluble DEAE-Sephadex
-
additional information
additional information
-
-
-
additional information
additional information
-
Michaelis-Menten kinetics using Lineweaver-Burk and Hanes plots at pH 8.5 and 22C, purified recombinant enzyme
-
additional information
additional information
-
kinetic analysis of the perhydrolytic wild-type and mutant enzyme activities, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.022
acetyl-L-Phe
Bacillus subtilis
-
30C, pH 8.0
0.38
acetyl-L-Phe ethyl ester
Bacillus subtilis
-
30C, pH 8.0
0.1
acetyl-L-Tyr
Bacillus subtilis
-
30C, pH 8.0
-
0.04
acetyl-L-Tyr ethyl ester
Bacillus subtilis
-
30C, pH 8.0
1900
benzyloxycarbonyl-Gly-p-nitroanilide
Bacillus sp.
-
pH 7.5, 20C
2500
benzyloxycarbonyl-Gly-p-nitroanilide
Bacillus sp.
-
pH 7.5, 20C, DTT
16550
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-4-nitroanilide
Bacillus intermedius
-
-
64.3
Glp-Ala-Ala-Leu-p-nitroanilide
Bacillus subtilis
-
in 2% DMF
69
Glp-Ala-Ala-Leu-p-nitroanilide
Bacillus licheniformis
-
in 2% DMF
80
Glp-Ala-Ala-Leu-p-nitroanilide
Bacillus subtilis
-
in 2% DMSO
35.5
Hammarsten casein
Bacillus clausii
-
37C, pH 10.5
-
31
L-Ala-L-Ala-L-Pro-L-Ala
Bacillus lentus
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
47
L-Ala-L-Ala-L-Pro-L-Ala
Bacillus lentus
-
wild-type, pH 9.0, 24C
185
L-Ala-L-Ala-L-Pro-L-Phe
Bacillus lentus
-
wild-type, pH 9.0, 24C
403
L-Ala-L-Ala-L-Pro-L-Phe
Bacillus lentus
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
10
L-Ala-L-Ala-L-Val-L-Ala
Bacillus lentus
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
12
L-Ala-L-Ala-L-Val-L-Ala
Bacillus lentus
-
wild-type, pH 9.0, 24C
90
L-Phe-L-Ala-L-Ala-L-Phe
Bacillus lentus
-
wild-type, pH 9.0, 24C
189
L-Phe-L-Ala-L-Ala-L-Phe
Bacillus lentus
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
4
L-Tyr-L-Val-L-Ala-L-Asp
Bacillus lentus
-
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24C
10
L-Tyr-L-Val-L-Ala-L-Asp
Bacillus lentus
-
wild-type, pH 9.0, 24C
47
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide
Bacillus sp.
-
subtilisin DY
57
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide
Bacillus amyloliquefaciens, Bacillus sp.
-
subtilisin Carlsberg
0.0064
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C
0.0074
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C, mutant D180G
0.0079
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C, mutant D182G
0.0089
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C; pH 8.5, 37C, mutant D180G/D182G
231.3
N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus amyloliquefaciens
-
pH 7.4, 37C, wild-type
586.2
N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus amyloliquefaciens
-
pH 7.4, 37C, mutant S101W/G169A/V192A
1.7
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant mutant T58A/G165I/L216W, pH 7.5, 30C
15.1
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant mutant T58A/G165L/L216W, pH 7.5, 30C
22.8
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant mutant T58A/G165Y/L216W, pH 7.5, 30C
33.1
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant mutant T58A/L216W, pH 7.5, 30C
172.6
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant wild-type enzyme, pH 7.5, 30C
419
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
pH 8.0, 25C, recombinant wild-type enzyme
462
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
pH 8.0, 25C, recombinant wmutant T211G
475
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
pH 8.0, 25C, recombinant mutant N212G
638
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
pH 8.0, 25C, recombinant mutant P210G
697
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
pH 8.0, 25C, recombinant mutant P210G/T211G
715
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
pH 8.0, 25C, recombinant mutant P210A
759
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
pH 8.0, 25C, recombinant mutant P210G/T211G/N212G
1.6
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
Thermococcus kodakarensis
-
subtilisin-like serine protease, at 20C, in 50 mM Tris-HCl buffer, pH 7.5
14
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
Thermococcus kodakarensis
-
subtilisin, at 20C, in 50 mM Tris-HCl buffer, pH 8.0
25
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
Thermococcus kodakarensis
-
subtilisin-like serine protease, at 80C, in 50 mM Tris-HCl buffer, pH 7.5
440
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
Thermococcus kodakarensis
-
subtilisin, at 80C, in 50 mM Tris-HCl buffer, pH 8.0
70.2
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide
Bacillus clausii
-
37C, pH 10.5
222
Suc-Ala-Ala-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 5C
750
Suc-Ala-Ala-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 20C, DTT
1200
Suc-Ala-Ala-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 20C
22000
Suc-Ala-Ala-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 40C
97000
Suc-Ala-Ala-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 75C
93
Suc-Ala-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 20C, in presence and absence of DTT
3700
Suc-Ala-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 40C
222
Suc-Ala-Ala-Pro-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 5C
8800
Suc-Ala-Ala-Pro-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 40C
130000
Suc-Ala-Ala-Pro-Ala-p-nitroanilide
Bacillus sp.
-
pH 7.5, 75C
389
Suc-Ala-Ala-Pro-Leu-p-nitroanilide
Bacillus sp.
-
pH 7.5, 5C
97000
Suc-Ala-Ala-Pro-Leu-p-nitroanilide
Bacillus sp.
-
pH 7.5, 40C
90
Suc-Ala-Ala-Pro-Phe-4-nitroanilide
synthetic construct
-
3.5 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25C
132
Suc-Ala-Ala-Pro-Phe-4-nitroanilide
synthetic construct
-
2.2 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25C
166
Suc-Ala-Ala-Pro-Phe-4-nitroanilide
synthetic construct
-
1 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25C
200
Suc-Ala-Ala-Pro-Phe-4-nitroanilide
synthetic construct
-
subtilisin without bound lactose, in 10 mM potassium phosphate buffer, pH 7.8, at 25C
148
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
Bacillus sp.
-
pH 7.5, 5C
2900
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
Bacillus sp.
-
pH 7.5, 20C, DTT
10000
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
Bacillus sp.
-
pH 7.5, 20C
130000
Suc-Ala-Ala-Pro-Phe-p-nitroanilide
Bacillus sp.
-
pH 7.5, 40C
0.052 - 2.1
succinyl-AAPA-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-g
6.8
succinyl-AAPA-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-c and S166C-S-e
9.65
succinyl-AAPA-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-g
17.7
succinyl-AAPA-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C wild-type
28.2
succinyl-AAPA-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-f
1.75
succinyl-AAPE-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C wild-type
14.5
succinyl-AAPE-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-a
0.017
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C-S-a
0.49
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C-S-c
0.833
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S166C-S-f
1.04
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C-S-f
1.64
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C-S-e
2.94
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C-S-e
3.7
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C-S-e
3.8
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S166C-S-e
4.8
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-f
6.08
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C-S-f
6.9
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S166C-S-d and M222C-S-d
7.2
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C-S-b
11.8
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S166C-S-b
14.2
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S166C
16.3
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S166C-S-f
16.9
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-c
17.3
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C
17.4
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S166C-S-a
18
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C-S-c
21.8
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C-S-d
23.1
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-c
23.4
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C-S-b
24.2
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C-S-c
25
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-b
29
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-g
30
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C-S-a
31
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S166C-S-c
31.6
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C-S-e
33.3
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant M222C-S-a
39
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C-S-f
43
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C mutant S156C
47
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-d
48
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 7.5, 25C, wild-type
48
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C, mutant S166C-h
50
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-a
67
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-i
76
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-j
82
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-e
153
succinyl-AAPF-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C wild-type
0.16
succinyl-AAPR-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C wild-type
0.17
succinyl-AAPR-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-b
0.35
succinyl-AAPR-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-h
0.43
succinyl-AAPR-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-i
0.68
succinyl-AAPR-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-d
1.06
succinyl-AAPR-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-j
6.08
succinyl-AAPR-p-nitroanilide
Bacillus lentus
-
pH 8.6, 25C mutant S166C-S-d; pH 8.6, 25C mutant S166C-S-j
1.6
Succinyl-Ala-Ala-Ala 4-nitroanilide
Bacillus sp.
-
subtilisin Carlsberg
1.8
Succinyl-Ala-Ala-Ala 4-nitroanilide
Bacillus sp.
-
subtilisin DY
52
succinyl-Ala-Ala-Phe 4-nitroanilide
Bacillus sp.
-
subtilisin DY
45
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide
Bacillus amyloliquefaciens
-
mutant subtilisin BPN' Asp99Ser
57
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide
Bacillus amyloliquefaciens, Bacillus sp.
-
subtilisin BPN'
6.4
succinyl-L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
Bacillus amyloliquefaciens
-
mutant Y217L, pH 8.3, 25C
15
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant wild-type enzyme, in presence of 2 M GdmCl
-
34
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant S62I/A153V/G166S/I205V, in presence of 2 M GdmCl
-
35
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant wild-type enzyme, in presence of SDS
-
42
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant S62I/A153V/G166S/I205V, in absence of GdmCl or SDS
-
45
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant S62I/A153V/G166S/I205V, in presence of SDS
-
50
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant wild-type enzyme, in absence of GdmCl or SDS
-
62
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant G166M, in presence of 2 M GdmCl
-
65
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant S62I, in presence of SDS
-
70
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant S62I/G166M, in presence of 2 M GdmCl
-
81
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant S62I/G166M, in presence of SDS
-
102
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant S62I, in absence of GdmCl or SDS
-
104
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant G166M, in presence of SDS
-
123
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant S62I/G166M, in absence of GdmCl or SDS
-
141
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide
Bacillus subtilis
-
pH 8.5, 37C, recombinant mutant G166M, in absence of GdmCl or SDS
-
210
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide
Bacillus amyloliquefaciens
-
mutant Y217L, pH 8.3, 25C
68
succinyl-L-Asp-L-Val-L-Arg-L-Ala-L-Phe-7-amido-4-methylcoumarin
Bacillus amyloliquefaciens
-
mutant Y217L, pH 8.3, 25C
29
succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl 7-amide
Bacillus sp.
-
subtilisin Sendai
0.04
Urea-denatured hemoglobin
Bacillus subtilis
-
30C, pH 8.0
-
70.9
Z-Ala-Ala-Leu-p-nitroanilide
Bacillus subtilis
-
in 20% DMF
77
Z-Ala-Ala-Leu-p-nitroanilide
Bacillus licheniformis
-
in 20% DMSO
100
Z-Ala-Ala-Leu-p-nitroanilide
Bacillus subtilis
-
in 20% DMSO
104
Z-Ala-Ala-Leu-p-nitroanilide
Bacillus licheniformis
-
in 20% DMF
97
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide
Bacillus sp.
-
subtilisin
additional information
additional information
Bacillus sp., Bacillus subtilis
-
-
-
additional information
additional information
Bacillus subtilis
-
-
-
additional information
additional information
Bacillus sp.
-
-
-
additional information
additional information
Bacillus sp.
-
turnover number of subtilisin Novo chemically attached to soluble DEAE-dextran and insoluble DEAE-Sephadex
-
additional information
additional information
synthetic construct
-
increasing the level of glycosylation causes a linearly dependent reduction in structural dynamics, which leads to a decrease in the catalytic turnover rate for both, the enzyme acylation and deacylation steps
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.67
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C
2798
1.73
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C, mutant D180G
2798
2.11
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C, mutant D182G
2798
2.21
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C, mutant D180G/D182G
2798
2.84
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
Dichelobacter nodosus
-
pH 8.5, 37C
2798
660
N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus amyloliquefaciens
-
pH 7.4, 37C, wild-type
19580
3910
N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus amyloliquefaciens
-
pH 7.4, 37C, mutant S101W/G169A/V192A
19580
3.1
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant mutant T58A/G165I/L216W, pH 7.5, 30C
953
36
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant mutant T58A/G165L/L216W, pH 7.5, 30C
953
114
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant mutant T58A/G165Y/L216W, pH 7.5, 30C
953
220.7
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant mutant T58A/L216W, pH 7.5, 30C
953
292.5
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide
Bacillus licheniformis
-
recombinant wild-type enzyme, pH 7.5, 30C
953
6
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
Thermococcus kodakarensis
-
subtilisin, at 20C, in 50 mM Tris-HCl buffer, pH 8.0
3147
15
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
Thermococcus kodakarensis
-
subtilisin-like serine protease, at 20C, in 50 mM Tris-HCl buffer, pH 7.5
3147
56
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
Thermococcus kodakarensis
-
subtilisin, at 80C, in 50 mM Tris-HCl buffer, pH 8.0
3147
62
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide
Thermococcus kodakarensis
-
subtilisin-like serine protease, at 80C, in 50 mM Tris-HCl buffer, pH 7.5
3147
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00274
2,2'-[[(1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphoryl]bis(oxybenzene-4,1-diyl)]diacetic acid
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
2,2'-[[(1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphoryl]bis(oxybenzene-4,1-diyl)]diacetic acid
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.00135
2,2'-[[(1-[[1-(tert-butoxycarbonyl)-L-prolyl]amino]-3-methylbutyl)phosphoryl]bis(oxybenzene-3,1-diyl)]diacetic acid
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(2,3,5-trimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(2,3-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(2,3-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(2,5-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(2-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(2-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(3,4,5-trimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(3,4,5-trimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(3,4-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(3,4-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(3-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(3-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(3-methoxyphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(4-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(4-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(4-ethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(4-ethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(4-methoxyphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(4-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis(4-tert-butylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.00792
bis(4-tert-butylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.000076
bis[4-(methylsulfonyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis[4-(propan-2-yl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis[4-(propan-2-yl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.00213
bis[4-(sulfanylmethyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
bis[4-(sulfanylmethyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.000000003
chymotrypsin inhibitor
-
-
-
0.000000017
chymotrypsin inhibitor 2 mutant M59A, chymotrypsin inhibitor 2 mutant M59F
-
-
-
0.00000013
chymotrypsin inhibitor 2 mutant M59G
-
-
-
0.000000019
chymotrypsin inhibitor 2 mutant M59K
-
-
-
0.0000000033
chymotrypsin inhibitor 2 mutant M59Y
-
-
-
0.00000046
chymotrypsin inhibitor 2 mutant Y61A
-
-
-
0.00637
diphenyl (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
diphenyl (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.0000043
EPI1a
-
predicted by use of Laskowski algorithm
-
50
EPI1b
-
predicted by use of Laskowski algorithm
-
0.0000000339
fungal protease inhibitor F
-
mutant T29M
-
0.000000148
fungal protease inhibitor F
-
wild-type
-
0.000000285
fungal protease inhibitor F
-
mutant T29L
-
0.000000327
fungal protease inhibitor F
-
mutant T29F
-
0.000000678
fungal protease inhibitor F
-
mutant T29E
-
0.00000245
fungal protease inhibitor F
-
mutant T29R
-
0.00000468
fungal protease inhibitor F
-
mutant T29G
-
0.000049
human LEKTI
-
pH 7.4, 25C
-
0.0000000036
human proteinase inhibitor 9
-
pH 7.5, 25C
-
0.000093
N-(tert-butoxycarbonyl)-L-valyl-N-(1-[bis[4-(sulfanylmethyl)phenoxy]phosphoryl]-2-phenylethyl)-L-prolinamide
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.00023
N-(tert-butoxycarbonyl)-L-valyl-N-[1-(diphenoxyphosphoryl)-2-phenylethyl]-L-prolinamide
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(3,4,5-trimethylphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.00006
N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(4-methoxyphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(4-tert-butylphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.001
N1-18[ISP]
-
synthetic peptide corresponding to the N-terminal extension behaves as a mixed noncompetitive inhibitor of active ISP, pH and temperature not specified in the publication
0.0025
RNA aptamer
-
pH 7.8, 37C
-
0.00206
Suc-Val-Pro-PheP(OPh)2
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
tert-butyl (2S)-2-([1-[bis(2-methylphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.02
tert-butyl (2S)-2-([1-[bis(3,4,5-trimethylphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.036
tert-butyl (2S)-2-([1-[bis(3,4-dimethylphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.00705
tert-butyl (2S)-2-([1-[bis(4-methoxyphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.00228
tert-butyl (2S)-2-([1-[bis(4-methylphenoxy)phosphoryl]-3-methylbutyl]carbamoyl)pyrrolidine-1-carboxylate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.0053
tert-butyl (2S)-2-[(1-[bis[4-(sulfanylmethyl)phenoxy]phosphoryl]-2-phenylethyl)carbamoyl]pyrrolidine-1-carboxylate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
0.0121
tert-butyl (2S)-2-[[1-(diphenoxyphosphoryl)-2-phenylethyl]carbamoyl]pyrrolidine-1-carboxylate
-
in 50 mM Tris, 1M NaCl, pH 7.5 containing 0.01% Triton X-100
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
700
guanidinium hydrochloride
Bacillus subtilis
-
pH 7.5, 37C, recombinant wild-type enzyme
960
guanidinium hydrochloride
Bacillus subtilis
-
pH 7.5, 37C, recombinant mutant N218S
1260
guanidinium hydrochloride
Bacillus subtilis
-
pH 7.5, 37C, recombinant mutant T224A
2730
guanidinium hydrochloride
Bacillus subtilis
-
pH 7.5, 37C, recombinant mutant S62I/A153V/G166S/I205V
3070
guanidinium hydrochloride
Bacillus subtilis
-
pH 7.5, 37C, recombinant mutant S62I/A153V/G166S/T224A/T240S
4650
guanidinium hydrochloride
Bacillus subtilis
-
pH 7.5, 37C, recombinant mutant S62I/A153V/G166S/I205V/N218S/T224A
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.02
-
40C, pH 7.5, solvent: hexane
0.23
-
40C, pH 7.5, solvent: diethanolammonium chloride
6
-
with N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide as substrate at pH 7.0
28.1
-
pH 8.0, 30C
80
-
40C, pH 7.5, solvent: buffer K2HPO4/KH2PO4
115.9
toward collagen
510
-
subtilisin-like serine protease
696
-
enzyme purified 1480fold
3100
-
subtilisin
4357
-
pH 10.0, 30C
6852
-
37C, pH 10.5
additional information
-
assay procedure for esterolytic ,subtilisin Carlsberg or subtilisin BPN', and proteolytic activity
additional information
-
specific activity of purified protease 174 units/mg, 1 unit = DELTA A420 of 1.0 per min at 75C with 0.2% azocasein
additional information
-
strain DJ-4, specific activity 0.98
additional information
-
easy and flexible green fluorescence protein based assay for enzymic activity, which is performed under almost authentic washing conditions
additional information
-
the virulent protease BprV is more efficient than its sequence homolog BprB
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7
recombinant enzyme
7 - 8
-
denatured hemoglobin, subtilisin Novo, subtilisin BPN'
7 - 8
broad
7.5
recombinant prosubtilisin JB1, at 37C
7.5
-
assay at
8.5
-
substrate casein
8.5
-
assay at
8.5 - 9
in 100 mM glycine-NaOH buffer
8.6
GN111900.1
-
9
-
substrate benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-4-nitroanilide
10
-
succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide
10
-
strain DJ-4
10
-
with beta-casein
10 - 11
-
casein, subtilisin Carlsberg
10.5
-
native subtilisin and its modified counterparts
10.7
-
subtilisin Novo immobilized on porous glass with the aid of hexamethylene diisocyanate
additional information
-
alteration of pH-profile by protein engineering
additional information
-
mutant subtilisin Asp to Ser99, change of just one surface charge which is 14-15 A from the active site of subtilisin has a significant effect on the pH-dependence
additional information
-
-
additional information
-
pI: greater than 9.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
3.5 - 11
-
native subtilisin and its modified counterparts
4 - 11
-
strain DJ-4
4 - 8
recombinant prosubtilisin JB1, at 37C
6 - 10
-
over 60% of maximum activity at pH 6-10, with a sharp decline in activity above pH 10, at 50C, profile overview
6 - 9
-
protease activity decreases significantly below pH 6 and above pH 9
7 - 11
-
at pH 7.0, 75% of maximum activity
7 - 11
-
optimal reaction
7 - 11.5
-
subtilisin-like serine protease: pH 7.0-11.5, subtilisin: pH 8.0 and 11.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
-
assay with human fibronectin
25
-
assay at
37
-
assay with N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide
37
-
assay at
37
-
assay at
37
-
assay at
40
-
strain DJ-4
40
recombinant enzyme
45
at pH 9.5 in 100 mM glycine-NaOH buffer
50
-
succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl 7-amide
55
-
in presence of Ca2+
60
-
in absence of Ca2+
60
-
recombinant enzyme
60
GN111900.1
-
60 - 65
-
subtilisin Novo immobilized on porous glass with the aid of hexamethylene diisocyanate
67
-
native subtilisin and its modified counterparts
70
-
in presence of 5 mM Ca2+
100
-
subtilisin-like serine protease: 100C, subtilisin: 90C
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
15 - 65
-
activity range, profile overview
20 - 80
-
G56S-propeptide is almost completely degraded within 510 min at 80C, 1530 min at 60C and 3 h at 40C, but is not completely degraded upon incubation for 3 h at 20C. Pro-subtilisin is rapidly autoprocessed into propeptide and mature subtilisin within 1 min even at 20C. However, the subsequent degradation of propeptide proceeds much more slowly than that of G56S-propeptide. Propeptide is almost completely degraded only when it is incubated at 80C for 30 min. It is not completely degraded for up to 3 h at 20, 40 and 60C
30
-
about 40% of maximum activity
30 - 60
-
at 30C, 78% of maximum activity
30 - 70
-
40% of maximal activity at 30C and 70C, 70% at 50C, at pH 8.6, profile overview
45 - 50
-
optimal reaction
additional information
-
mesophilic
additional information
-
thermophilic
additional information
-
cold adapted, negatively charged residues on the surface may be responsible for cold adaptation
additional information
cold adapted, negatively charged residues on the surface may be responsible for cold adaptation
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.91
sequence calculation
8
-
isoelectric focusing, KV42
8.19
-
isoelectric focusing, KV42
8.4
-
isoelectric focusing
8.62
-
isoelectric focusing, KV71
8.8
-
determined in granulated dextran gels
8.8
-
isoelectric focusing
8.83
-
isoelectric focusing, KV71
8.92
-
isoelectric focusing, KV42
8.98
-
isoelectric focusing, KV42
9.1
-
isoelectric focusing, KV71
9.2
-
isoelectric focusing
9.4
-
subtilisin Carlsberg
9.47
-
isoelectric focusing, KV71 and KV42
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
from Sigma; subtilisin Carlsberg: from Sigma
Manually annotated by BRENDA team
-
from Novo Industries; subtilisin Carlsberg: from Sigma
Manually annotated by BRENDA team
-
from Boehringer; from Serva
Manually annotated by BRENDA team
-
the commercial enzyme subtilsin BPN' from Sigma: kinetic and physical properties indicate that the commercial enzyme is probably subtilisin Carlsberg isolated from Bacillus licheniformis, much work which has been performed on commercial subtilisin BPN' from 1975 up to 1985 must be revaluated
Manually annotated by BRENDA team
-
subtilisin Carlsberg: from Sigma
Manually annotated by BRENDA team
-
bacterial enzyme preparation, Alcalase from Novo Industries; subtilisin Novo: crude enzyme preparation (Alcalase 2.0T) from Novo
Manually annotated by BRENDA team
-
from Novo Industries
Manually annotated by BRENDA team
-
from Enzyme Development Corporation, New York
Manually annotated by BRENDA team
Bacillus intermedius Mrz 19
-
-
-
Manually annotated by BRENDA team
mRNA levels and enzyme activity increase during leaf senescence in leaves senescing during both the vegetative or the reproductive phase of the plant life cycle, but this increase is more pronounced in reproductive plants
Manually annotated by BRENDA team
additional information
the enzyme is expressed in all aboveground organs, but not in roots
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
immunofluorescence shows SUB1 on the trichomonad surface, and in dispersed vesicles throughout the cytoplasm
Manually annotated by BRENDA team
-
extracellular in healthy tissues, shifts to cytoplasm upon programmed cell death induction
-
Manually annotated by BRENDA team
Bacillus sp. DJ-4, Bacillus sp. G-825-6, Fusarium equiseti CBS119568, Trichoderma harzianum ACCC30371
-
-
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacillus amyloliquefaciens subsp. plantarum (strain DSM 23117 / BGSC 10A6 / FZB42)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Thermococcus kodakarensis (strain ATCC BAA-918 / JCM 12380 / KOD1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
16900
-
SDS-PAGE
717205
27000
-
subtilisin BPN', sedimentation equilibrium measurement
29427
27000
-
-
717697
27230
-
subtilisin Sendai, amino acid sequence
29459
27280
-
subtilisin Carlsberg, amino acid sequence
29427
27290
-
subtilisin Novo, determination from amino acid composition
29425
27400
-
subtilisin Carlsberg, sedimentation equilibrium measurement
29427
27500
-
subtilisin amylosacchariticus, sedimentation equilibrium measurement
29429
27500
-
Bacillus stearothermophilus NCIMB10278 gene expressed in Bacillus subtilis DB104/pZS101
29478
27530
-
Bacillus sp., subtilisin BPN', determination from amino acid composition
29425
27670
-
Bacillus sp., subtilisin amylosacchariticus, amino acid sequence
29427
27800
calculated from the sequence
652805
28000
-
Bacillus sp., subtilisin amylosacchariticus, sedimentation equilibrium measurement
29427
29000
-
strain DJ-4, SDS-PAGE
650609
31000
-
-
649830
31550
-
mass spectroscopy, processed form
718327
34780
-
MALDI-TOF
718405
36000
-
SDS-PAGE
649692
37000
-
sequence analysis or mass spectrometry
707232
38000
SDS-PAGE
718103
41000
-
SDS-PAGE
718118
41300
calculated online with ProtParam tools
707878
42000
calculated online with ProtParam tools
707878
42000
calculated online with ProtParam tools
707878
42000
calculated online with ProtParam tools
707878
44000
-
Pro-subtilisin-like serine protease derivative with N- and/or C-terminal truncation, gel filtration
710470
44200
-
subtilisin-like serine protease, sequence analysis
710470
45000
-
pro-S255A, gel filtration
684164
55000
-
Pro-subtilisin-like serine protease derivative with N- and/or C-terminal truncation, gel filtration
710470
55000
-
dimer, gel filtration
718405
62000
purified prosubtilisin JB1, Western blotting analysis
707215
65000
-
Pro-subtilisin-like serine protease, gel filtration
710470
68630
-
Pro-subtilisin-like serine protease, sequence analysis
710470
74500
-
MALDI-TOF
717456
80000
-
single polypeptide chain
717456
82000
Western blot, mature form
718185
84000
-
SDS-PAGE
717456
101000
calculated fromc DNA
718185
169600
C5VW36
sequence analysis
707888
170000
-
SDS-PAGE
718443
additional information
-
amino acid sequence of: subtilisin BPN'
29425
additional information
-
X-ray structure
29426
additional information
-
amino acid composition of: subtilisin BPN'; subtilisin amylosacchariticus; subtilisin Carlsberg; subtilisin Novo
29427
additional information
-
amino acid sequence of: subtilisin BPN'; subtilisin amylosacchariticus; subtilisin Carlsberg; subtilisin DY
29428
additional information
-
characterization of a folding intermediate
29446
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 28000, Bacillus sp., SDS-PAGE
?
-
x * 25000, Bacillus sp., SDS-PAGE
?
-
x * 28000, SDS-PAGE
?
-
x * 27000, SDS-PAGE
?
x * 29000, mature enzyme
?
-
x * 26500, SDS-PAGE
?
-
x * 31000, SDS-PAGE, x * 27636, mass spectrometry
?
x * 55000, SDS-PAGE, x * 65000, SDS-PAGE
?
-
x * 29000, native enzyme, SDS-PAGE, x * 29141, sequence calculation, x * 29140, recombinant enzyme, mass spectrometry
?
GN111900.1
x * 27000, recombinant enzyme, SDS-PAGE
?
-
x * 34000, SDS-PAGE
?
x * 59000-61000, zymogen enzyme form, DS-PAGE
?
x * 44900, about, sequence calculation, x * 69000, recombinant enzyme, SDS-PAGE
?
Alkalimonas collagenimarina AC40T
-
x * 55000, SDS-PAGE, x * 65000, SDS-PAGE
-
?
Bacillus clausii GMBAE 42
-
x * 26500, SDS-PAGE
-
?
Bacillus gibsonii DSM 14391
-
x * 27000, recombinant enzyme, SDS-PAGE
-
?
Bacillus intermedius Mrz 19
-
x * 28000, SDS-PAGE
-
?
-
x * 29000, mature enzyme
-
?
-
x * 28000, Bacillus sp., SDS-PAGE
-
?
Bacillus sp. GX6644
-
x * 25000, Bacillus sp., SDS-PAGE
-
?
Bacillus subtilis CN2
-
x * 31000, SDS-PAGE, x * 27636, mass spectrometry
-
?
Bacillus subtilis IBTC-3
-
x * 27000, SDS-PAGE
-
?
Fusarium equiseti CBS119568
-
x * 29000, native enzyme, SDS-PAGE, x * 29141, sequence calculation, x * 29140, recombinant enzyme, mass spectrometry
-
?
Trichoderma harzianum ACCC30371
-
x * 44900, about, sequence calculation, x * 69000, recombinant enzyme, SDS-PAGE
-
dimer
-
MALDI-TOF, 2 * 34782 Da
monomer
-
1 * 29000, strain DJ-4, SDS-PAGE
monomer
-
-
monomer
-
pro-S255A, SDSPAGE
monomer
-
1 * 27000, SDS-PAGE
monomer
-
1* 42000, gel filtration, subtilisin-like serine protease. 1 * 65000, SDS-PAGE, Pro-S359A
monomer
-
SDS-PAGE, 1 * 16900 Da
monomer
-
1 * 29000, strain DJ-4, SDS-PAGE
-
monomer
Bacillus subtilis TA41
-
-
-
additional information
-
N-terminal amino acid sequence
additional information
-
N-terminal and internal amino acid sequencing, overview
additional information
-
N-terminal amino acid sequence
-
additional information
Fusarium equiseti CBS119568
-
N-terminal and internal amino acid sequencing, overview
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
-
propeptide is absolutely required to fold into a kinetically trapped conformer. Comparison of folding kinetics and intermediates with intracellular serine proteases
proteolytic modification
-
sites of primary autoproteolysis of the purified recombinant Fe protease are Leu2-Thr3, Gly11-Leu12, Trp143-Ala144, Ala173-Ser174, Ala179-Asn180, and Trp219-Tyr220 (numbered according to the amino acids in the mature protease)
proteolytic modification
Fusarium equiseti CBS119568
-
sites of primary autoproteolysis of the purified recombinant Fe protease are Leu2-Thr3, Gly11-Leu12, Trp143-Ala144, Ala173-Ser174, Ala179-Asn180, and Trp219-Tyr220 (numbered according to the amino acids in the mature protease)
-
proteolytic modification
-
expressed as pre-pro-proteins whose prodomains are autocatalytically processed
proteolytic modification
the enzyme matures from the inactive precursor, Pro-Tk-subtilisin (Pro-TKS), upon autoprocessing and degradation of the propeptide (Tkpro)
proteolytic modification
in silico analyses predicts signal and pro-peptides at the N-terminus
additional information
occurrence of the two variant forms of AtSASP can be due to posttranslational modifications
proteolytic modification
-
expressed as pre-pro-proteins whose prodomains are autocatalytically processed
additional information
-
contains no carbohydrate
proteolytic modification
-
prosequence regulates ISP activity through two distinct modes: active site blocking and catalytic triad rearrangement. The full-length proenzyme is inactive until specific proteolytic processing removes the first 18 amino acids that comprise the N-terminal extension, with processing appearing to be performed by ISP itself
additional information
-
contains no carbohydrate
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure at 2.4 A resolution
-
1.8 A resolution structure of an inactive form (by replacing the catalytic nucleophile Ser 221 with alanine) of the protease subtilisin S189, in complex with azide and with a prodomain substrate that spans the active site. The substrate is well ordered across the active site, and the azide anion is observed bound adjacent to Ala 32. Although S189, like wild-type subtilisin, has Ser as the catalytic nucleophile at residue 221, these crystal structures have Ala 221 to prevent cleavage of the substrate
-
in complex with chymotrypsin inhibitor 2 and its mutants M59K, M59Y, M59F, M59A, M59G, Y61A
-
wild-type and thermostable mutant 7150 subtilisin at 1.8 A resolution
-
structure of the processed protein is determined at 2.6 A resolution and compared with that of the full-length protein, in which the N-terminal extension binds back over the active site. A conserved proline introduces a backbone kink that shifts the scissile bond beyond reach of the catalytic serine and in addition the catalytic triad is disrupted. In the processed form, access to the active site is unblocked by removal of the N-terminal extension and the catalytic triad rearranges to a functional conformation
-
subtilisin Carlsberg covalently bound to the inhibitor L-[(1R)-1-acetamido-2-(1-naphthyl)ethyl]boronic acid, refined at 2.65 A is used in the studies; the structure is determined to a resolution of 1.56 A. ISP from Bacillus clausii is dimeric, with residues from the C terminus making a major contribution to the dimer interface by crossing over to contact the partner subunit. A short N-terminal extension binds back across the active site to provide a potential novel regulatory mechanism of intrinsic proteolytic activity: a proline residue conserved throughout the ISPs introduces a kink in the polypeptide backbone that lifts the target peptide bond out of reach of the catalytic residues
-
at 1.4 A resolution
-
crystal structure at 1.4 A resolution
-
hanging drop vapor diffusion, crystals belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell dimensions of 52.65 A, 61.25 A and 74.75 A
-
subtilisin DY in a 1:1 complex with the synthetic inhibitor N-benzyloxycarbonyl-Ala-Pro-Phe-chloromethyl ketone, space group P2(1)2(1)2(1), unit cell dimensions of a = 5.28 nm, b = 7.27 nm and c = 5.98 nm
-
trans-cinnamoyl-subtilisin, space group P2(1)2(1)2(1), unit cell dimensions in H2O a = 77.0, b = 55.0, c = 53.6, unit cell dimensions in acetonitrilea = 76.4 A, b = 55.3 A and c = 52.8 A
-
crystal structure at 2.5 A
-
crystal structure determined by molecular replacement, crystals grown in hanging drops, monoclinic space group P2(1), with cell dimensions a : 44.1 A, b : 51.7 A, c : 52.8 A, coordinates and structure factor amplitudes in the ProteinDataBank codes 1DBI and R1DBISF
crystal structure of a subtilisin BPN' complex with N-benzoyl-L-Arg
-
crystal structure of subtilisin BPN' with Streptomyces subtilisin inhibitor at 2.6 A and 4.3 A
-
crystal structure of subtilisin E with PMSF inhibitor at 2 A resolution
-
crystallographic study
-
refined 1.2 A crystal structure of the complex formed between subtilisin Carlsberg and the inhibitor eglin c
-
refined crystal structure of complex of subtilisin BPN' and Streptomyces subtilisin inhibitor at 1.8 A resolution
-
refined crystal structures of subtilisin Novo in complex with wild-type and two mutant eglins
-
structure at 2.5 A resolution
-
X-ray analysis
-
crystal structure of subtilisin E with PMSF inhibitor at 2 A resolution
-
subtilisin S41 surface is rich in hydrophilic residues, particularly Asp, which may contribute to its adaptation to psychrophilic conditions
-
X-ray analysis
-
by the hanging-drop vapour-diffusion method. Crystals of AprB2 diffract to 1.7 A resolution. The crystals belong to space group P1, with unit-cell parameters a = 42.7, b = 45.8, c = 45.7 A , alpha = 98.4, beta = 114.0, gamma = 114.6. The crystals contain one molecule in the asymmetric unit, with a solvent content of 36%; by the hanging-drop vapour-diffusion method. Crystals of AprV2 diffract to 2.0 A resolution. The crystals belong to space group P1, with unit-cell parameters a = 43.1, b = 46.0, c = 47.2 A , alpha = 97.8, beta = 115.2, gamma = 115.2. The crystals contain one molecule in the asymmetric unit, with a solvent content of ca. 36%; by the hanging-drop vapour-diffusion method. Crystals of BprB diffract to 1.8 A resolution. The crystals belong to space group P21, with unit-cell parameters a = 38.5, b = 90.5, c = 44.1 A , beta = 109.9. The crystals contain one molecule in the asymmetric unit, with a solvent content of 36%; by the hanging-drop vapour-diffusion method. Crystals of BprV diffract to 2.0 A resolution. The crystals belong to space group P21, with unit-cell parameters a = 38.5, b = 89.6, c = 47.7 A , beta = 113.6. The crystals contain one molecule in the asymmetric unit, with a solvent content of 40%
crystal structure shows that an unusual extended disulfhide-tethered loop functions as an exosite, mediating effective enzyme-substrate interactions. The disulphide bond and Tyr92, which is located at the exposed end of the loop, is functionally important
-
data reveal that that the S1 pocket of BprB is less hydrophobic but bigger than that of basic protease BprV (sequence homologue); data reveal that that the S1 pocket of BprV is more hydrophobic but smaller than that of basic protease BprB
-
by the batch method from buffer solution saturated with Na2SO4 ca. 13% (w/v) as precipitant. Placement of several Cs+ and Cl- ions in crystals of subtilisin Carlsberg. The protein conformation is very similar to that of the enzyme without CsCl in acetonitrile. 11 defined sites for Cs+ cations and 8 Cl- anions around the protein molecule, although most of these have partial occupancy and may represent nonspecific binding sites. Two Cs+ and two Cl- ions are close to the mouth of the active site cleft, where they may affect catalysis. CsCl-treated subtilisin crystals transferred to acetonitrile show catalytic activity several fold higher than the reference crystals containing Na+
by the hanging drop vapor-diffusion method, at 0.8 A resolution. Crystals belong to the monoclinic space group P21. Unit cell parameters are a = 46.09 A, b = 62.67 A, c = 84.87 A, and beta = 95.5, indicating two molecules of Sph in the asymmetric unit. The crystal structures of the psychrophilic Bacillus TA41 subtilisin S41 and the mesophilic subtilisin Sph are nearly identical with the same calcium-loaded state in that five calcium ions are bound to each protein molecule; by the hanging drop vapor-diffusion method, at 1.4 A resolution. Crystals belong to either the tetragonal space group P41212 or P43212. Unit cell dimensions are a = b = 61 A and c = 174.77 A , indicating one monomer in the asymmetric unit. The crystal structures of the psychrophilic subtilisin S41 and the mesophilic Bacillus sphaericus subtilisin Sph are nearly identical with the same calcium-loaded state in that five calcium ions are bound to each protein molecule
serine protease inhibitor CrSPI-1 complexed with subtilisin. Crystals diffracted to 2.6 A resolution and belong to space group P2(1), with unit-cell parameters a = 73.8, b = 65.0, c = 111.9 A, beta = 95.4. Presence of one heterotrimer (1:2 ratio of CrSPI-1:subtilisin) and one free subtilisin molecule in the asymmetric unit
synthetic construct
-
a comparative study of psychrophilic, mesophilic and thermophilic subtilisin-like serine proteases by all-atom molecular dynamics simulations is performed. The thermophilic subtilisin presents a high affinity calcium binding site which is not structurally conserved in the mesophilic and psychrophilic counterparts, which at the same position show a stable salt bridge network and no stabilizing intra-molecular interactions, respectively
-
by sitting-drop, vapor-diffusion method. Crystal structures of the autoprocessed/unautoprocessed form and mature forms of subtilisin at 1.89 A and 1.70 A resolution, respectively. Crystals show a unique Ca2+-binding site and that the N-terminal region of the mature domain (Gly70Pro82), which binds tightly to the main body in the unautoprocessed form, is disordered and mostly truncated in the autoprocessed and mature forms, respectively. Crystal structure of the propeptide:S324A-subtilisin complex at 1.65 a resolution
-
by the sitting drop vapor diffusion method. Crystal structures of propeptides in complex with S324A-subtilisin. Conformation of the propeptide is altered by the mutation, such that nonglycine residues at position 56 assume a right-handed conformation and hydrophobic interactions at the core region decrease
-
pro-S255A crystallized in complex with Ca2+, by sitting-drop vapour-diffusion method, to 2.3 A resolution. Crystal belongs to space group I222, with unit cell parameters a = 92.69, b = 121.78, c = 77.53 A. Matthews coefficient is 2.6 A 3 Da-1 and the solvent content is 53.1%.
-
high content of acidic residues mainly found on its surface, making it charged
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
1 - 2
-
denaturation
29425
1 - 3
-
24 h, 76% loss of activity
669738
2 - 12
-
subtilisin-like serine protease is fully stable between pH 7-11, whereas it is not fully stable below pH 6 and above pH 12. It loses more than 85% of its activity below pH 3 and at pH 13. Subtilisin is fully stable between pH 2-12
710470
3 - 11
-
stable within this range
669822
4
-
both active and denatured subtilisins are present simultaneously, rapid autolysis
29425
4
recombinant enzyme, about 85% residual activity after 48 h at 4C, native enzyme, 60% residual activity
668657
4 - 12
-
enzyme loses its activity at pH lower 4, retains 60% of its activity at pH higher 12
717697
4.5
-
24 h, complete loss of activity
29466
5.5 - 10
-
24 h, stable
29466
5.5 - 9
-
stable
29458
6.3 - 9.5
-
-
667756
9 - 12.2
-
30C, stable for 24 h
669738
11
-
above, destabilization of enzyme structure
29425
12.7
-
24 h, 38% loss of activity
669738
additional information
-
subtilisin Novo immobilized on porous glass with the aid of hexamethylene diisocyanate is stable in a wider range of pH than the native enzyme
29480
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
22 - 45
-
presence of Ca2+
667756
30 - 40
-
pH 10.5, stable for 2 h
669738
37
-
relatively unstable when incubated at 37C for 18 h (80% reduction)
718443
40
residual activity is 80%, 60%, or 3% of the original activity after 15 min incubation at 45C, 50C, or 55C, respectively. CaCl2 slightly increases the thermal stability of AcpII up to 45C, and the residual activity is decreased to 80% after incubation at 50C for 15 min
707260
45
-
heating the recombinant protease at 45C results in a 20% reduction of activity
718443
50
-
3 h, 30% loss of activity, thermostable strain
29431
50
-
stable below
29458
50
-
strain DJ-4, 80% of the enzyme's activity remains after incubation for 1 h
650609
50
-
pH 10.5, 2 h, 85% residual activity
669738
50
-
30 min, stable
669822
50
-
remarkable loss of activity at temperature higher than 50C
717697
55
-
rapid denaturation above
29425
55
-
stable up to
671007
60
-
30 min, presence of 2 mM calcium, about 50% loss of activity
29478
60
recombinant enzyme, stable for 40 min, native enzyme, 30% loss of activity after 40 min
668657
60
-
30 min, 85% residual activity
669822
60
-
30 min, 85% resiudal activity
671007
62 - 71
synthetic construct
-
increasing the level of glycosylation causes a linearly dependent reduction in structural dynamics, which leads to an increase in thermostability: subtilisin without bound lactose: 64C, 1 mol lactose bound per 1 mol subtilisin: 62C, 2.2 mol lactose bound per 1 mol subtilisin: 69C and 3.5 mol lactose bound per 1 mol subtilisin: 71C
707805
65
-
pH 8.0, 10 mM CaCl2, half-life of wild-type enzyme: 59 min, half-life of mutant 7150 enzyme: 223 min
29450
65
-
pH 10.0, 10 min, stable up to
29459
65
-
heating the recombinant protease at 65C results in a 65% reduction of activity
718443
70
-
pH 10.0, 10 min, complete loss of activity
29459
70
-
30 min, 20% residual activity
669822
70 - 105
highly thermostable, heat-treating for 2 h at 70C activates the protease, half-life at 85C in presence of DTT is 34 min, in absence of DTT 317 min, at 105C complete loss of activity is virtually instantaneous in absence of sorbitol, in presence of 90% sorbitol the half-life is 104 min
652805
80
-
subtilisin-like serine protease is stable at 80C for at least 3. It loses activity at 90C and 100C with half-lives of more than 3 h and 100 min, respectively. Subtilisin loses its activity at 100C with a half-life of 50 min
710470
100
-
heating the recombinant protease at 100C results in a total loss of activity
718443
additional information
-
Ca2+ stabilizes the enzyme upon heat treatment
29478
additional information
-
subtilisin Novo immobilized on porous glass with the aid of hexamethylene diisocyanate is stable in a wider range of temperature than the native enzyme
29480
additional information
the high structural similarity between Bacillus sphaericus Sph and S41 and their identical calcium-binding mode, indicates that the calcium-loaded state is not responsible for the cold adaptation of S41
710507
additional information
requirement of Ca2+ ions for the hyperthermostability of Tk-subtilisin.The Ca1, Ca6, and Ca7 ions, especially the Ca1 ion, contribute to the hyperthermostabilization of Tk-subtilisin
726988
additional information
GN111900.1
thermostability profiles of purified wild-type enzyme, and mutants N253D, Q256E, and N253D/Q256E, overview
731201
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Alteration of stability to oxidative, thermal and alkaline inactivation by protein engineering
-
stability is a property of subtilisin which has proven particularly amenable to enhancement via random mutagenesis
-
subtilisin is readily capable of degrading itself
-
Relatively resistant to denaturation by guanidine solutions
-
Urea, 4 M, no effect
-
stability of wild-type an dmutant enzymes in the presence of GdmCl or SDS, overview
-
Substrate binding stabilizes the enzyme
-
Urea, 8 M, only slow loss of ordered structure and caseinolytic activity
-
Ca2+ stabilizes the enzyme upon heat treatment
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2-methyl-2-propanol
-
effect on synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2, yield of dipeptide is 56.9% at 85% 2-methyl-2-propanol
acetonitrile
-
effect on synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2, yield of dipeptide is 75.5% at 85% acetonitrile
acetonitrile
-
flexibility of subtilisin Carlsberg decreases during prolonged storage. Is more flexible in acetonitrile than in 1,4-dioxane
chloroform
-
effect on synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2, yield of dipeptide is 51.3% at 85% chloroform
dimethylformamide
-
effect on synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2, yield of dipeptide is 33.55% at 85% dimethylformamide
dioxane
-
in 1,4-dioxane, flexibility of subtilisin Carlsberg decreases during prolonged storage: incubation of lyophilized enzyme for a maximum of 96 h (4 days) in 1,4-dioxane, a solvent in which this enzyme is most active and enantioselective, shows a decrease in the rate of exchange of both fast and slow exchanging protons
Ethanol
-
effect on synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2, yield of dipeptide is 49.4% at 85% ethanol
Ethyl acetate
-
effect on synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2, yield of dipeptide is 43.7% at 85% ethyl acetate
Methanol
-
effect on synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2, yield of dipeptide is 16.3% at 85% methanol; effect on synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2, yield of dipeptide is 33.55% at 85% methanol
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by ammonium sulfate precipitation, to almost homogeneity, 364fold with recovery of 24.9% of the original activity. AcpII-C and AcpII-C-DELTAPA purified 109- and 219fold, with overall yields of 6.9% and 0.36%, respectively
using Ni-NTA chromatography
-
by affinity purification, more than 95% pure
-
subtilisin BPN' and recombinant variants
-
using a HiTrap Q HP column
-
from culture filtrate
-
using Ni-NTA chromatography
-
recombinant wild-type and mutant enzymes from Bacillus subtilis strain DB104 by dialysis and anion and cation exchange chromatography
GN111900.1
wild-type and mutant enzymes
-
by ammonium sulfate precipitation followed by gel filtration
-
overview: isolation procedure
-
purification by single-step affinity chromatography of a crude enzyme preparation (Alcalase 2.0T) from Novo
-
recombinant enzyme
-
strain DJ-4
-
strain TA41
-
by gel filtration
-
prosubtilisin purified by ammonium sulfate precipitation and gel filtration
purification of pro-subtilisin E wild-type and mutant (Asp32 to Asn) and intramolecular autoprocessing to active subtilisin E in vitro
-
purified to 23.5fold by a combination of cation and anion exchange chromatography, ethanol precipitation followed by reverse-phase HPLC
-
recombinant wild-type and mutant enzymes from Bacillus subtilis strain WB600 culture supernatant by anion and cation exchange chromatography and ultrafilatration
using Ni-NTA chromatography
by affinity chromatography; by affinity chromatography; by affinity chromatography; by affinity chromatography
native extracellular enzyme from culture supernatant by anion exchangc chromatography, ammonium sulfate fractionation, hydrophobic interaction chromatography, ultrafiltration, and gel filtration, recombinant enzyme from Trichoderma reesei
-
NCIMB10278, gene expressed in Bacillus subtilis DB104/pZS101
-
by ion-exchange chromatography
-
to homogeneity by ion exchange chromatography and gel filtration. Alkaline proteolytic activity enriched 1480fold with a yield of 27%
-
native enzyme from culture filtrate by ultrafiltration, anion exchange and hydrophobic interaction chromatography, to homogeneity
-
using ion-exchange chromatography, ultrafiltration and gel filtration
-
G56S-propeptide and His-tagged G56S-propeptide purified by gel filtration
-
G56W-propeptide and G56E-propeptide, S324A-subtilisin purified in denatured form
-
mutants purified in a urea-denatured form
-
pro-S255A purified in the presence of 8 M urea. Refolded by removing urea in the presence of Ca2+. Further purified by gel filtration
-
S324A-subtilisin and propeptide, by gel filtration
-
subtilisin-like serine protease and Pro-S359A, at 4C, by sonication, centrifugation, ammonium sulfate precipitation and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
vectors pRSET-acpII, pRSET-acpII-tr (truncated AcpII Met1-Arg675), pRSET-acpII-C (catalytic domain of AcpII) and pRSET-acpII-C-DELTAPA (AcpII-C with the protease-associated domain deleted) expressed in Escherichia coli BL21(DE3) pLysS
gene At3g14067, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression in an enzyme-knockout line sasp-1 or SALK_147962 and functional complementation
overexpressed in Escherichia coli as a His-tagged fucion protein
-
almost every property of subtilisin has been altered by protein engineering, e.g. substrate specificity, pH-profile and stability to oxidative, thermal and alkaline inactivation
-
Bacillus amyloliquefaciens gene expressed in Bacillus subtilis I-168 (Marburg strain)
-
expressed in the protease deficient host Bacillus subtilis ISW1214
-
in vivo formation and stability of engineered disulfide bonds in subtilisin
-
inactive mutants of subtilisin BPN expressed in Bacillus subtilis. Expression of prodomain mutant proR9 in Escherichia coli strain BL21(DE3)
-
subtilisin 7150, mutagenized subtilisin with enhanced thermal stability
-
subtilisin BPN' cloned, sequenced and expressed at high levels from its natural promoter sequences in Bacillus subtilis
-
subtilisin variants expressed in Escherichia coli
-
expressed in Escherichia coli
-
expression of wild-type and mutant enzymes in Bacillus subtilis strain DB104, subcloning in Escherichia coli strain DH5alpha
GN111900.1
cloning in Escherichia coli strain DH5alpha and Saccharomyces cerevisiae strain YPH499, recombinant expression of wild-type and mutant enzymes in Bacillus megaterium strain WH320 from a shuttle expression vector carrying a ribosome-binding site and a signal sequence of extracellular metalloprotease from Bacillus megaterium downstream of xylose isomerase promoter
-
recombinant expression of mutant enzyme sin Bacillus subtilis strain DB104, subcloning in Escherichia coli strain DH5alpha
-
AkP gene cloned and expressed in Escherichia coli PB5517
-
expression of full-length enzyme and mature form, expression of mature form does not give enzymatic activity
replacement of the Bacillus subtilis subtilisin structural gene with an in vitro-derived deletion mutant
-
subtilisin Carlsberg from Bacillus licheniformis expressed in Bacillus subtilis
-
subtilisin E produced in Escherichia coli
-
aprE gene rraprE
-
gene aprE, construction of error-prone PCR libraries, recombinant expression of wild-type and mutant enzymes in Bacillus subtilis strain WB600 and secretion to the culture medium
-
gene aprE, recombinant expression of wild-type and mutant enzymes in Bacillus subtilis strain WB600 and secretion to the culture medium
-
prosubtilisin JB1 cloned into the GST fusion protein expression vector pGEX-4T-1 and transformed into Escherichia coli DH5alphaMCR
replacement of the Bacillus subtilis subtilisin structural gene with an in vitro-derived deletion mutant
-
subtilisin E produced in Escherichia coli
-
subtilisin gene from Bacillus subtilis PTCC 1023 is cloned into the vector pET-15b and expressed in Escherichia coli strain BL21
vector pET22b-AprB2 encoded residues 1-474 of the AprB2 precursor with a C-terminal His6 transformed into Escherichia coli DH5alpha. Expression construct transformed into Escherichia coli strain Rosetta-Gami(DE3)pLysS; vector pET22b-AprV2 encoded residues 1-474 of the AprV2 precursor with a C-terminal His6 tag transformed into Escherichia coli strain Rosetta-Gami(DE3)pLysS; vector pET22b-BprB encoded residues 1-476 of BprB with a C-terminal His6 tag transformed into Escherichia coli strain Rosetta-Gami(DE3)pLysS; vector pET22b-BprV encoded residues 1-476 of BprV with a C-terminal His6 tag and transformed into Escherichia coli strain Rosetta-Gami(DE3)pLysS
DNA and amino acid sequence determination and analysis
-
gene prtS8A, DNA and amino acid sequence determination and analysis, fusion to the Trichoderma reesei cbh1 (cel7A) promoter and recombinant expression in Trichoderma reesei
-
Bacillus stearothermophilus NCIMB10278 gene expressed in Bacillus subtilis DB104/pZS101
-
full-length SUB2 gene from strain GS115 transformed into Escherichia coli. Overexpressed in Pichia pastoris KM71
-
cloned and amino acid sequence deduced from its cDNA
-
DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain DH5alpha
-
cloned and expressed in Escherichia coli
-
Escherichia coli HB101 transformed with plasmid pBR1560 containing the wild-type prepro-subtilisin. G56S-propeptide and His-tagged G56S-propeptide overexpressed in Escherichia coli BL21-CodonPlus(DE3)-RIL transformed with vector pET25b and in Escherichia coli BL21(DE3) with vector pET28a, respectively
-
overexpression of pro-S255A by pET25b in Escherichia coli BL21-Codon-Plus(DE3)
-
pET25b derivatives for overproduction of Pro-G56W, Pro-G56E, G56W-propeptide, and G56E-propeptide. G56W-propeptide overproduced in soluble form, while G56E-propeptide overproduced in insoluble form. S324A-subtilisin overproduced in inclusion bodies
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prosubtilisn, DELTAloop-subtilisin, DELTACa2-Pro-S324A, and DELTACa3-Pro-S324A cloned into pET25b vector and overexpressed in Escherichia coli in inclusion bodies
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the gene encoding the 1689 derivative without a putative N-terminal signal sequence, Pro-subtilisin-like serine protease (Met + Ala1-Gly640), ligated into pET25b vector, mutant Pro-S359A overexpressed in Escherichia coli BL21-CodonPlus(DE3)-RIL cells
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gene ThSS45, DNA and amino acid sequence determination and analysis, phylogenetic tree, quantitative expression analysis, recombinant expression in in Escherichia coli strain BL21
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
transcription of gene ThSS45 is up-regulated when grown in mineral medium, under carbon starvation, and nitrogen starvation, and in the presence of 1% root powder (3.5fold), 1% stem powder, and 1% leaf powder derived from Populus davidiana P. bolleana (Shanxin poplar) aseptic seedlings. Gene transcription of ThSS45 is also slightly up-regulated by 1% Alternaria alternata cell wall and 5% Alternaria alternata fermentation liquid
transcription of gene ThSS45 is up-regulated when grown in mineral medium, under carbon starvation, and nitrogen starvation, and in the presence of 1% root powder (3.5fold), 1% stem powder, and 1% leaf powder derived from Populus davidiana P. bolleana (Shanxin poplar) aseptic seedlings. Gene transcription of ThSS45 is also slightly up-regulated by 1% Alternaria alternata cell wall and 5% Alternaria alternata fermentation liquid
Trichoderma harzianum ACCC30371
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transcript abundance is found to be modulated by the availability of iron in the growth medium
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A116E
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site-directed mutagenesis, BPN'
A194P
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site-directed mutagenesis, savinase
A1C
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does not change the catalytic properties of subtilisin S189 but allows the introduction of a fluorescent group at its N-terminus
D32A
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engineered variant of the protease subtilisin, denoted S189, mutation renders the enzyme's activity dependent on the presence of certain small anions such as fluoride or azide. Is activated more than 3000fold by azide
D32N
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does not fold at all in the presence of proR9
D97G
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site-directed mutagenesis, subtilisin E
E89S
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site-directed mutagenesis, BPN'
G131D
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site-directed mutagenesis, BPN'
G166R
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site-directed mutagenesis, subtilisin E
G166S
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site-directed mutagenesis, BPN'
G169A
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stable only in presence of excess Ca2+
G169A
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site-directed mutagenesis, BPN'
G169A
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mutant shows 1.4fold activity compared to wild-type
G195E
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site-directed mutagenesis, BPN'
H120D
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site-directed mutagenesis, savinase
I107V
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site-directed mutagenesis, BPN'
K213R
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site-directed mutagenesis, BPN'
K235L
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site-directed mutagenesis, savinase
K256Y
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site-directed mutagenesis, BPN'
K27R
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site-directed mutagenesis, BPN'
L126I
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site-directed mutagenesis, BPN', stable only in presence of excess Ca2+
L126I
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site-directed mutagenesis, BPN'
M50F
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site-directed mutagenesis, BPN',stabilized in EDTA, but destabilized slightly in CaCl2
N109S
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site-directed mutagenesis, BPN'
N118S
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site-directed mutagenesis, subtilisin E
N155L
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folding kinetics very similar to that of S221A
N181D
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site-directed mutagenesis, subtilisin E
N218D
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site-directed mutagenesis, BPN'
N218S
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site-directed mutagenesis, BPN', stabilized significantly, either with or without Ca2+
N218S
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site-directed mutagenesis, BPN'; site-directed mutagenesis, subtilisin E
N76D
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site-directed mutagenesis, BPN'; site-directed mutagenesis, subtilisin E
N77K
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site-directed mutagenesis, BPN'
P14L
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site-directed mutagenesis, subtilisin E
P168G
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weaker binding of proR9
P172D
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site-directed mutagenesis, BPN'
Q19E/Q271E
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site-directed mutagenesis, BPN'
Q206Cox
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site-directed mutagenesis, BPN'
Q271E
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site-directed mutagenesis, BPN', stabilized significantly, either with or without Ca2+
Q271E
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site-directed mutagenesis, BPN'
R170Y
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site-directed mutagenesis, BPN'
S101K/G169A
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mutant shows 3.3fold activity compared to wild-type
S101L/G169A
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mutant shows 2.9fold activity compared to wild-type
S101R/G169A
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mutant shows 2.2fold activity compared to wild-type
S101W/G169A
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mutant shows 3.9fold activity compared to wild-type
S101W/G169A/V192A
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mutant shows 1.4fold activity compared to wild-type, kcat doubled compared to wild-type, Km decreased compared to wild-type
S128G
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site-directed mutagenesis, savinase
S161C
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site-directed mutagenesis, subtilisin E
S188P
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site-directed mutagenesis, BPN', stable only in presence of excess Ca2+
S188P
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site-directed mutagenesis, BPN'
S194P
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site-directed mutagenesis, BPN'; site-directed mutagenesis, subtilisin E
S221A
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a delta75-83 mutant. Shows slow folding reaction. Is unstable at low metal concentration, unfolding at a rate of 0.6 h-1. If denatured, it is refolded with an excess of proR9
S221A
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inactive form of subtilisin S189
S221C
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site-directed mutagenesis, BPN'
S221C
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folding kinetics very similar to that of S221A
S53TI
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site-directed mutagenesis, BPN'
S63D
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site-directed mutagenesis, BPN'
S78D
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site-directed mutagenesis, BPN'
T22C/S87C/S221C
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if denatured, it is refolded with an excess of proR9. About 75% is rapidly bound to proR9. 25% undergoes a slow step prior to binding
T254A
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site-directed mutagenesis, BPN', stable only in presence of excess Ca2+
T254A
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site-directed mutagenesis, BPN'
V205I
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site-directed mutagenesis, savinase
V8I
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site-directed mutagenesis, BPN'
Y217K
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site-directed mutagenesis, BPN', stabilized significantly, either with or without Ca2+
Y217K
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site-directed mutagenesis, BPN'
Y217L
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commercial version of enzyme, i.e. SBT*
N115D
GN111900.1
site-directed mutagenesis, inactive mutant
N154D
GN111900.1
site-directed mutagenesis, the mutant has 107% proteolytc activity compared to the wild-type enzyme
N167D
GN111900.1
site-directed mutagenesis, the mutant has 77% proteolytc activity compared to the wild-type enzyme
N205D
GN111900.1
site-directed mutagenesis, the mutant has 113% proteolytc activity compared to the wild-type enzyme
N236D
GN111900.1
site-directed mutagenesis, the mutant has 115% proteolytc activity compared to the wild-type enzyme
N242D
GN111900.1
site-directed mutagenesis, the mutant has 118% proteolytc activity compared to the wild-type enzyme
N250D
GN111900.1
site-directed mutagenesis, the mutant has 113% proteolytc activity compared to the wild-type enzyme
N253D
GN111900.1
site-directed mutagenesis, the mutant has 135% proteolytc activity compared to the wild-type enzyme and shows a dramatic pH activity profiles shifted towards higher activity at lower pH range of pH 8.5-10
N253D/Q256E
GN111900.1
site-directed mutagenesis, the mutant shows a dramatic pH activity profiles shifted towards higher activity at lower pH range of pH 8.5-10, the mutant has a 2fold increased activity compared to the wild-type enzyme with a thermal resistance increased by 2.4C at pH 8.5
N60D
GN111900.1
site-directed mutagenesis, the mutant has 32% proteolytc activity compared to the wild-type enzyme
N97D
GN111900.1
site-directed mutagenesis, the mutant has 146% proteolytc activity compared to the wild-type enzyme
Q107E
GN111900.1
site-directed mutagenesis, the mutant has 53% proteolytc activity compared to the wild-type enzyme
Q12E
GN111900.1
site-directed mutagenesis, the mutant has 114% proteolytc activity compared to the wild-type enzyme
Q176E
GN111900.1
site-directed mutagenesis, the mutant has 97% proteolytc activity compared to the wild-type enzyme
Q185E
GN111900.1
site-directed mutagenesis, inactive mutant
Q200E
GN111900.1
site-directed mutagenesis, the mutant has 125% proteolytc activity compared to the wild-type enzyme
Q230E
GN111900.1
site-directed mutagenesis, the mutant has 107% proteolytc activity compared to the wild-type enzyme
Q256E
GN111900.1
site-directed mutagenesis, the mutant has 129% proteolytc activity compared to the wild-type enzyme and shows a dramatic pH activity profiles shifted towards higher activity at lower pH range of pH 8.5-10
N250D
Bacillus gibsonii DSM 14391
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site-directed mutagenesis, the mutant has 113% proteolytc activity compared to the wild-type enzyme
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N97D
Bacillus gibsonii DSM 14391
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site-directed mutagenesis, the mutant has 146% proteolytc activity compared to the wild-type enzyme
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Q107E
Bacillus gibsonii DSM 14391
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site-directed mutagenesis, the mutant has 53% proteolytc activity compared to the wild-type enzyme
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Q12E
Bacillus gibsonii DSM 14391
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site-directed mutagenesis, the mutant has 114% proteolytc activity compared to the wild-type enzyme
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M222C
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site-directed mutagenesis combined with chemical modification
S156C
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site-directed mutagenesis combined with chemical modification
S166C
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site-directed mutagenesis combined with chemical modification
V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I
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mutant containing the highly flexible region of psychrophilic enzyme TA39 subtilisin. Mutant shows the same temperature optimum and pH-profile as wild-type, but higher specificity for substrate succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide and broader substrate specificity. Mutant has a decreased thermostability, but increased activity at low temperature
G165X
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site-directed mutagenesis, catalytic constants of subtilisin Carlsberg for perhydrolysis of methylpropionate, methylbutyrate and methylpentanoate are increased up to 3.5fold, 5.4fold and 5.5fold, respectively, while proteolysis is decreased up to 100fold for N-succinyl-Ala-Ala-Pro-Phe-pnitroanilide substrate
N212G
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site-directed mutagenesis, the mutant shows increased kcat compared to the wild-type enzyme
P210A
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site-directed mutagenesis, the mutant shows increased kcat compared to the wild-type enzyme
P210G
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site-directed mutagenesis, the mutant shows 1.5fold increased kcat compared to the wild-type enzyme
P210G/T211G
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site-directed mutagenesis, the mutant shows 1.7fold increased kcat compared to the wild-type enzyme
P210G/T211G/N212G
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site-directed mutagenesis, the mutant shows 1.8fold increased kcat compared to the wild-type enzyme
S161C
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the mutant brings increased stability to subtilisin compared to the wild-type
T211G
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site-directed mutagenesis, the mutant shows 1.8fold increased kcat compared to the wild-type enzyme
T58A/L216W
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site-directed mutagenesis, the mutant catalyzes in addition to its proteolytic activity the generation of peroxycarboxylic acids from corresponding esters in the presence of hydrogen peroxide
A153V
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site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, but does not show improved properties in chaotropic conditions
G166M
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site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and improved properties in chaotropic conditions
G166S
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site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and improved properties in chaotropic conditions
I205V
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site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, but does not show improved properties in chaotropic conditions
K211P/R212A
mutant is more thermostable compared to wild-type, half-life at 60C 10times longer compared to wild-type. Molecular dynamics simulation at 10C and 90C reveal that the average global flexibility of both variants is slightly higher than wild-type
N218S
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site-directed mutagenesis, the mutant shows increased activity and stability in the presence of GdmCl or SDS compared to the wild-type enzyme
S62I
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site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and slightly improved properties in chaotropic conditions
S62I/A153V/G166S/I205V
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site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and highly improved properties in chaotropic conditions
S62I/A153V/G166S/I205V
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site-directed mutagenesis, the mutant shows increased activity and stability in the presence of GdmCl or SDS compared to the wild-type enzyme
S62I/A153V/G166S/I205V/N218S/T224A
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site-directed mutagenesis, the mutant shows high activity and stability in the presence of GdmCl or SDS compared to the wild-type enzyme
S62I/A153V/G166S/T224A/T240S
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site-directed mutagenesis, the mutant shows high activity and stability in the presence of GdmCl or SDS compared to the wild-type enzyme; site-directed mutagenesis, the mutant shows increased activity and stability in the presence of GdmCl or SDS compared to the wild-type enzyme
S62I/G166M
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site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and highly improved properties in chaotropic conditions
T224A
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site-directed mutagenesis, the mutant shows increased activity and stability in the presence of GdmCl or SDS compared to the wild-type enzyme
D180G
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mutant using substrate N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide Km similar to wild-type, kcat value enhanced, kcat/Km value enhanced by 4%
D180G/D182G
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using substrate N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide Km similar to wild-type, kcat value highly enhanced, kcat/Km value enhanced by 33%
D182G
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using substrate N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide Km similar to wild-type, kcat value enhanced, kcat/Km value enhanced by 27%
DELTA83-99
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truncation mutant is not functional
Y92A
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mutant shows decreased elastin degradation
Y92D
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mutant shows decreased elastin degradation
Y92F
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mutant shows increased elastin degradation
Y92L
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mutant shows decreased elastin degradation
G6G
C5VW36
possesses a single Tn917 insertion, is devoid of subtilisin-like proteinase activity, has longer generation times and is more susceptible to killing by whole blood than the wild-type
M3G
C5VW36
possesses a single Tn917 insertion, is devoid of subtilisin-like proteinase activity, has longer generation times and is more susceptible to killing by whole blood than the wild-type
G6G
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possesses a single Tn917 insertion, is devoid of subtilisin-like proteinase activity, has longer generation times and is more susceptible to killing by whole blood than the wild-type
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