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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
Q0WWH7;
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
-
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 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
malfunction
Q0WWH7;
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
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
-
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
A1YN98;
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
-
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
-
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
Q0WWH7;
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
-
AcpII comprises a prepropeptide, a catalytic domain that includes a protease-associated domain, and tandem repeat prepeptidase C-terminal domains
-
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
-
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
-
the serine protease may be involved in both mycoparasitism and antibiotic secretion
-
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
Q0WWH7;
occurrence of the two variant forms of AtSASP can be due to posttranslational modifications
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
(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
-
-
-
-
-
(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
-
-
-
-
-
(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
-
-
-
-
-
67 kDa gamma-glutamyl transpeptidase + H2O
30 kDa gamma-glutamyl transpeptidase + ?
-
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
?
-
-
-
-
?
acetyl-L-Ala-L-Ala-L-Ala-4-nitroanilide + H2O
?
-
-
-
-
?
acetyl-L-Phe + ethanol
acetyl-L-Phe ethyl ester + H2O
acetyl-L-Phe ethyl ester + H2O
acetyl-L-Phe + ethanol
acetyl-L-Tyr + ethanol
acetyl-L-Tyr ethyl ester + H2O
-
-
-
-
r
acetyl-L-Tyr ethyl ester + H2O
acetyl-L-Tyr + ethanol
acid casein + H2O
?
-
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-methylcoumarin + H2O
?
Ala-Ala-Pro-Leu-4-nitroanilide + H2O
Ala-Ala-Pro-Leu + 4-nitroaniline
-
low activity
-
-
?
Ala-Ala-Pro-Lys-4-nitroanilide + H2O
Ala-Ala-Pro-Lys + 4-nitroaniline
-
low activity
-
-
?
Ala-Ala-Pro-Met-4-nitroanilide + H2O
Ala-Ala-Pro-Met + 4-nitroaniline
-
low activity
-
-
?
Ala-Ala-Pro-Nle-4-nitroanilide + H2O
Ala-Ala-Pro-Nle + 4-nitroaniline
-
low activity
-
-
?
Ala-Ala-Pro-Phe 4-nitroanilide + H2O
?
Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Ala-Ala-Pro-Phe + 4-nitroaniline
-
low activity
-
-
?
Ala-Ala-Val-Ala-4-nitroanilide + H2O
Ala-Ala-Val-Ala + 4-nitroaniline
-
low activity
-
-
?
Benzoyl-Arg ethyl ester + H2O
?
benzoyl-L-Arg + ethanol
benzoyl-L-Arg ethyl ester + H2O
-
-
-
-
r
Benzoyl-L-Arg ethyl ester + H2O
Benzoyl-L-Arg + ethanol
Benzyloxycarbonyl-Ala-Ala-Leu 4-nitroanilide + H2O
?
-
-
-
-
-
Benzyloxycarbonyl-Gly-Gly-L-Leu-NH2 + H2O
?
-
-
-
-
-
Benzyloxycarbonyl-Gly-L-Leu-NH2 + H2O
?
-
-
-
-
-
Benzyloxycarbonyl-Gly-L-Tyr-NH2 + H2O
?
-
-
-
-
-
Benzyloxycarbonyl-glycyl-L-tyrosinamide + H2O
?
-
-
-
-
-
benzyloxycarbonyl-L-Ala ethyl ester + butanol
benzyloxycarbonyl-L-Ala butyl ester + ethanol
-
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
benzyloxycarbonyl-L-Asp methyl ester + serine amide
benzyloxycarbonyl-L-Asp-L-Ser-NH2 + methanol
-
-
-
-
?
beta subunit of haemoglobin + H2O
?
-
-
-
-
?
Bovine serum albumin + H2O
?
Carboxybenzoyl-Gly-Ala-NH2 + H2O
Carboxybenzoyl-Gly-Ala + NH3
-
subtilisin BPN'
subtilisin BPN'
-
Carboxybenzoyl-Gly-Leu-NH2 + H2O
Carboxybenzoyl-Gly-Leu + NH3
-
subtilisin BPN'
subtilisin BPN'
-
Carboxybenzoyl-Gly-Pro-Leu-Gly-Pro-OH + H2O
Carboxybenzoyl-Gly-Pro-Leu + Gly-Pro-OH
-
subtilisin BPN'
subtilisin BPN'
-
chicken-feather keratin + H2O
?
-
-
-
-
?
Clupein + H2O
?
-
-
-
-
-
D-Val-Leu-Lys p-nitroanilide + H2O
D-Val-Leu-Lys + p-nitroaniline
-
-
-
?
eglin-c + H2O
?
-
proteinase inhibitor, subtilisin DY hydrolyses the peptide bond between residues 45 and 46 in the reactive centre of eglin-c
-
?
ethyl 2-(4-ethylphenoxy)propionate + butanol
butyl 2-(4-ethylphenoxy)propanoate + propanol
-
in isooctane, preferential use of S-enantiomer. Comparison with enantioselectivity in water and DMSO
-
-
?
Glp-Ala-Ala-Leu-p-nitroanilide + H2O
?
glutaryl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O
glutaryl-Ala-Ala-Pro-Leu + p-nitroaniline
relative hydrolysis rate is 22%
-
-
?
Hammarsten casein + H2O
?
Human fibrinogen + H2O
?
-
fibrinolytic activity is determined using the fibrin plate method
-
-
?
human fibronectin + H2O
?
-
-
-
-
?
insoluble elastin + H2O
?
-
-
-
-
?
kappa casein + H2O
?
-
purified caseins from animal's milk from cow, sheep, goat and water buffalo used as substrate for subtilisin
-
?
L-Ala-L-Ala-L-Ala-L-Ala-L-Pro-L-Phe + H2O
?
-
-
-
-
?
L-Ala-L-Ala-L-Phe-L-Ala-L-Ala-L-Phe + H2O
?
-
-
-
-
?
L-Ala-L-Ala-L-Pro-L-Ala + H2O
?
-
-
-
-
?
L-Ala-L-Ala-L-Pro-L-Phe + H2O
?
-
-
-
-
?
L-Ala-L-Ala-L-Val-L-Ala + H2O
?
-
-
-
-
?
L-Lys-L-His-L-Asp-L-Arg-L-Lys-L-Asp + H2O
?
-
-
-
-
?
L-Phe-L-Ala-L-Ala-L-Phe + H2O
?
-
-
-
-
?
L-phenylalanine-isopropylester + H2O
L-phenylalanine + isopropanol
-
-
-
-
-
L-Tyr-L-Val-L-Ala-L-Asp + H2O
?
-
-
-
-
?
lactate dehydrogenase + H2O
?
-
poor substrate in native form, but can be digested in heat-denatured form
-
-
?
malate dehydrogenase + H2O
?
-
poor substrate in native form, but can be digested in heat-denatured form
-
-
?
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide + H2O
?
methoxysuccinyl-Ala-Ile-Pro-Met-p-nitroanilide + H2O
methoxysuccinyl-Ala-Ile-Pro-Met + p-nitroaniline
is the most favorable substrate, relative hydrolysis rate is 100%
-
-
?
methyl mandelate + butanol
butyl mandelate + methanol
-
in isooctane, preferential use of S-enantiomer. Comparison with enantioselectivity in water and DMSO
-
-
?
N-Acetyl-Ala methyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-Acetyl-Gly ethyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-Acetyl-L-norvaline ethyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-acetyl-L-phenylalanine ethyl ester
?
-
-
-
-
?
N-acetyl-L-phenylalanine ethyl ester + 1-butanol
?
-
transesterification catalyzed by poly(ethylene glycol)-modified subtilisin
-
-
?
N-acetyl-L-phenylalanine ethyl ester + H2O
?
-
-
-
-
?
N-Acetyl-Leu methyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-Acetyl-Lys methyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-Acetyl-Phe ethyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-Acetyl-Phe methyl ester + H2O
?
N-Acetyl-Trp ethyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-Acetyl-Trp methyl ester + H2O
?
N-Acetyl-Tyr ethyl ester + H2O
?
N-acetyl-Tyr ethyl ester + H2O
N-acetyl-Tyr + ethanol
-
-
-
?
N-Acetyl-Tyr methyl ester + H2O
?
N-Acetyl-Val methyl ester + H2O
?
N-acetylglucosamine + H2O
?
N-CBZ-Gly-Gly-Leu p-nitroanilide + H2O
?
-
-
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide + H2O
N-methoxysuccinyl-Ala-Ala-Pro-Val + 4-nitroanilide
-
-
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide + H2O
N-methoxysuccinyl-Ala-Ala-Pro-Val + p-nitroaniline
N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
N-Succ-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
?
N-succinyl-Ala-Ala-Pro-Leu-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Leu + 4-nitroaniline
-
-
-
-
?
N-succinyl-Ala-Ala-Pro-Met-p-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Met + p-nitroaniline
relative hydrolysis rate is 39%
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
?
-
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
?
-
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
N-succinyl-Ala-Ala-Val-Ala-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Val-Ala + 4-nitroaniline
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-L-Ala-4-nitroanilide + H2O
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide + H2O
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
N-succinyl-L-Ala-L-Ala-L-Val-4-nitroanilide + H2O
?
N-tosyl-L-Arg methyl ester + H2O
N-tosyl-L-Arg + methanol
N-trans-cinnamoyl imidazole + H2O
(E)-cinnamate + imidazole
-
-
-
-
?
Nalpha-benzoyl-DL-Arg-4-nitroanilide + H2O
Nalpha-benzoyl-DL-Arg + 4-nitroaniline
-
-
-
-
?
Nalpha-Benzoyl-L-Arg ethyl ester + H2O
?
-
-
-
-
-
Oxidized insulin B-chain + H2O
?
oxidized insulin chain B + H2O
?
-
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
-
catalyzed by native subtilisin
-
-
?
peanut flour + H2O
?
-
6 h alcalase hydrolysate
-
-
?
peptidyl-4-nitroanilide + H2O
?
-
-
-
?
Phe-Ala-Ala-Phe-4-nitroanilide + H2O
Phe-Ala-Ala-Phe + 4-nitroaniline
-
highly preferred substrate
-
-
?
porcine brain tubulin + H2O
peptides
-
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
-
-
-
-
?
sec-phenethyl alcohol + vinyl butyrate
?
-
-
-
-
?
serum albumin + H2O
?
-
-
-
-
?
silk fibroin + H2O
?
-
-
-
-
?
Spirulina platensis powder + H2O
?
-
-
-
-
?
strobilurin fungicide + H2O
?
-
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-methyl-coumaryl-7-amide + H2O
?
-
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
succinyl-AAPA-p-nitroanilide + H2O
?
succinyl-AAPE-p-nitroanilide + H2O
?
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
succinyl-AAPF-p-nitroanilide + H2O
succinyl-alanyl-alanyl propyl-phenylalanine
-
-
-
?
succinyl-AAPR-p-nitroanilide + H2O
?
-
-
-
?
Succinyl-Ala-Ala 4-nitroanilide + H2O
?
Succinyl-Ala-Ala-Ala 4-nitroanilide + H2O
?
succinyl-Ala-Ala-Ala-Ala-Ala-p-nitroanilide + H2O
succinyl-AAAAA + p-nitroaniline
-
-
-
?
succinyl-Ala-Ala-Ala-p-nitroanilide + H2O
succinyl-AAA + p-nitroaniline
-
-
-
?
succinyl-Ala-Ala-p-nitroanilide + H2O
succinyl-Ala-Ala + p-nitroaniline
-
-
-
?
succinyl-Ala-Ala-Pro-Ala-p-nitroanilide + H2O
succinyl-AAPA + p-nitroaniline
-
-
-
?
succinyl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O
succinyl-AAPL + p-nitroaniline
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
?
succinyl-Ala-Ala-Pro-Phe-7-amido-4-methylcoumarin + H2O
succinyl-Ala-Ala-Pro-Phe + 7-amino-4-methylcoumarin
-
-
-
-
?
succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
-
-
-
-
?
succinyl-Ala2-Phe-4-methylcoumarin 7-amide + H2O
?
-
-
-
?
succinyl-L-Ala-L-Ala-L-Ala-4-nitroanilide + H2O
?
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
succinyl-L-Ala-L-Ala-L-Phe + 7-amino-4-methylcoumarin
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-Gly-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-Gly + 4-nitroaniline
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Ala-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Ala + 4-nitroaniline
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Lys-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Lys + 4-nitroaniline
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Met-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Met + 4-nitroaniline
-
-
-
-
?
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
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Phe + 4-nitroaniline
-
-
-
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Tyr-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Tyr + 4-nitroaniline
-
-
-
-
?
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
-
-
-
-
?
Succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide + H2O
?
succinyl-Phe-Ala-Ala-Phe-p-nitroanilide + H2O
succinyl-Phe-Ala-Ala-Phe + p-nitroaniline
-
-
-
?
Toluenesulfonyl-Arg methyl ester + H2O
?
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
type I keratin + H2O
?
-
-
-
-
?
Urea-denatured hemoglobin + H2O
?
vinyl butyrate + H2O
?
-
-
-
-
?
Z-Ala-Ala-Leu-OCH3 + H2O
?
-
-
-
-
?
Z-Ala-Ala-Leu-OMe + Phe-p-nitroanilide
Z-Ala-Ala-Leu-Phe-p-nitroanilide + methanol
-
-
-
-
?
Z-Ala-Ala-Leu-p-nitroanilide + H2O
?
Z-Ala-Phe-OMe + H2O
Z-Ala-Phe-NH2 + methanol
-
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 30°C 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
-
-
-
-
?
additional information
?
-
acetyl-L-Phe + ethanol

acetyl-L-Phe ethyl ester + H2O
-
-
-
-
r
acetyl-L-Phe + ethanol
acetyl-L-Phe ethyl ester + H2O
-
-
-
-
r
acetyl-L-Phe ethyl ester + H2O

acetyl-L-Phe + ethanol
-
-
-
-
r
acetyl-L-Phe ethyl ester + H2O
acetyl-L-Phe + ethanol
-
-
-
-
r
acetyl-L-Tyr ethyl ester + H2O

acetyl-L-Tyr + ethanol
-
-
-
-
r
acetyl-L-Tyr ethyl ester + H2O
acetyl-L-Tyr + ethanol
-
-
-
-
r
Ala-Ala-Phe 7-amido-4-methylcoumarin + H2O

?
-
-
-
-
?
Ala-Ala-Phe 7-amido-4-methylcoumarin + H2O
?
B6VFQ8;
-
-
-
?
Ala-Ala-Phe 7-amido-4-methylcoumarin + H2O
?
B6VFQ8;
-
-
-
?
Ala-Ala-Pro-Phe 4-nitroanilide + H2O

?
-
-
-
-
-
Ala-Ala-Pro-Phe 4-nitroanilide + H2O
?
-
-
-
-
-
azocasein + H2O

?
B6VFQ8;
-
-
-
?
azocasein + H2O
?
B6VFQ8;
-
-
-
?
azocasein + H2O
?
-
-
-
-
?
azocasein + H2O
?
-
-
-
-
?
Benzoyl-Arg ethyl ester + H2O

?
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
Benzoyl-Arg ethyl ester + H2O
?
-
subtilisin BPN', subtilisin Amylosacchariticus
-
-
-
Benzoyl-L-Arg ethyl ester + H2O

Benzoyl-L-Arg + ethanol
-
-
-
-
r
Benzoyl-L-Arg ethyl ester + H2O
Benzoyl-L-Arg + ethanol
-
-
-
-
r
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-4-nitroanilide + H2O

benzyloxycarbonyl-L-Ala-L-Ala-L-Leu + 4-nitroaniline
-
-
-
-
?
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-4-nitroanilide + H2O
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu + 4-nitroaniline
-
-
-
-
?
beta-casein + H2O

?
-
from bovine milk
-
-
?
beta-casein + H2O
?
-
from bovine milk
-
-
?
Bovine serum albumin + H2O

?
-
poor substrate in native form, but can be digested in heat-denatured form
-
-
?
Bovine serum albumin + H2O
?
-
-
-
-
?
Bovine serum albumin + H2O
?
B6VFQ8;
-
-
-
?
Bovine serum albumin + H2O
?
B6VFQ8;
-
-
-
?
Bovine serum albumin + H2O
?
-
-
-
-
?
Bovine serum albumin + H2O
?
-
-
-
-
?
Bovine serum albumin + H2O
?
-
-
-
-
?
casein + H2O

?
38% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on casein
-
-
?
casein + H2O
?
38% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on casein
-
-
?
casein + H2O
?
-
caseinolytic activity is determined using the casein plate method
-
-
?
chitin + H2O

?
-
-
-
-
?
Collagen + H2O

?
81% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on collagen
-
-
?
Collagen + H2O
?
81% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on collagen
-
-
?
cuticle + H2O

?
-
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
-
-
?
cuticle + H2O
?
-
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
-
-
?
Cytochrome c + H2O

?
-
from equine heart
-
-
?
Cytochrome c + H2O
?
-
from equine heart
-
-
?
Elastin + H2O

?
7% activity relative to gelatin as substrate
-
-
?
Elastin + H2O
?
-
-
-
-
?
Elastin + H2O
?
-
-
-
-
?
Fibrin + H2O

?
-
strain DJ-4
-
?
Fibrin + H2O
?
-
strain DJ-4
-
?
Fibrinogen + H2O

?
rapid hydrolysis of Aalpha-, Bbeta- and gamma-chains. At very low concentration, no cleavage of gamma-chain
-
-
?
Fibrinogen + H2O
?
rapid hydrolysis of Aalpha-, Bbeta- and gamma-chains. At very low concentration, no cleavage of gamma-chain
-
-
?
Fibrinogen + H2O
?
B6VFQ8;
-
-
-
?
Fibrinogen + H2O
?
B6VFQ8;
-
-
-
?
Gelatin + H2O

?
best substrate for AcpII. Also purified AcpII-C and AcpII-C-DELTAPA act well on gelatin
-
-
?
Gelatin + H2O
?
best substrate for AcpII. Also purified AcpII-C and AcpII-C-DELTAPA act well on gelatin
-
-
?
Gelatin + H2O
?
-
-
-
-
?
Gelatin + H2O
?
B6VFQ8;
-
-
-
?
Gelatin + H2O
?
B6VFQ8;
-
-
-
?
Gelatin + H2O
?
-
-
-
-
?
Glp-Ala-Ala-Leu-p-nitroanilide + H2O

?
-
-
-
-
?
Glp-Ala-Ala-Leu-p-nitroanilide + H2O
?
-
-
-
-
?
Glp-Ala-Ala-Leu-p-nitroanilide + H2O
?
-
-
-
-
?
Hammarsten casein + H2O

?
-
activation energy for hydrolysis 10.59 kcal per mol
-
-
?
Hammarsten casein + H2O
?
-
activation energy for hydrolysis 10.59 kcal per mol
-
-
?
Hemoglobin + H2O

?
-
-
-
-
-
Hemoglobin + H2O
?
-
denatured
-
-
-
Hemoglobin + H2O
?
-
-
-
-
?
Hemoglobin + H2O
?
-
-
-
-
?
Keratin + H2O

?
1.5% activity relative to gelatin as substrate
-
-
?
Keratin + H2O
?
1.5% activity relative to gelatin as substrate
-
-
?
Keratin + H2O
?
-
-
-
-
?
KOH chitin + H2O

?
-
-
-
-
?
KOH chitin + H2O
?
-
-
-
-
?
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide + H2O

?
-
-
-
-
-
MeO-succinyl-Ala-Ala-Phe 4-nitroanilide + H2O
?
-
-
-
-
-
N-Acetyl-Phe methyl ester + H2O

?
-
subtilisin Amylosacchariticus
-
-
-
N-Acetyl-Phe methyl ester + H2O
?
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Trp methyl ester + H2O

?
-
subtilisin Amylosacchariticus
-
-
-
N-Acetyl-Trp methyl ester + H2O
?
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Tyr ethyl ester + H2O

?
-
-
-
-
-
N-Acetyl-Tyr ethyl ester + H2O
?
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Tyr ethyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-Acetyl-Tyr methyl ester + H2O

?
-
subtilisin Amylosacchariticus
-
-
-
N-Acetyl-Tyr methyl ester + H2O
?
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Val methyl ester + H2O

?
-
subtilisin Carlsberg, subtilisin Novo
-
-
-
N-Acetyl-Val methyl ester + H2O
?
-
subtilisin BPN'
-
-
-
N-acetylglucosamine + H2O

?
-
restricted feeding of N-acetylglucosamine (the monomer of chitin) to biomass of KV71 induces protease to 61% of that on cuticle
-
-
?
N-acetylglucosamine + H2O
?
-
N-acetylglucosamine has little effect on subtilisin production by KV01 (11% of that on cuticle)
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide + H2O

N-methoxysuccinyl-Ala-Ala-Pro-Val + p-nitroaniline
-
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide + H2O
N-methoxysuccinyl-Ala-Ala-Pro-Val + p-nitroaniline
-
-
-
?
N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide + H2O
N-methoxysuccinyl-Ala-Ala-Pro-Val + p-nitroaniline
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O

N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O

N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
relative hydrolysis rate is 94%
-
-
?
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-L-Ala-4-nitroanilide + H2O

?
-
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-L-Ala-4-nitroanilide + H2O
?
-
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

?
-
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-Leu-4-nitroanilide + H2O
?
-
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-Leu-4-nitroanilide + H2O
?
-
-
-
-
-
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O

?
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide + H2O
?
-
-
-
-
?
N-succinyl-L-Ala-L-Ala-L-Val-4-nitroanilide + H2O

?
-
2.2% of the activity with N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide
-
-
?
N-succinyl-L-Ala-L-Ala-L-Val-4-nitroanilide + H2O
?
-
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
-
-
-
-
-
N-tosyl-L-Arg methyl ester + H2O
N-tosyl-L-Arg + methanol
-
subtilisin Carlsberg, subtilisin Novo, subtilisin Amylosacchariticus
-
-
-
Oxidized insulin B-chain + H2O

?
-
enzyme primarily hydrolyzes Leu15-Tyr16 bond and secondarily Gln4-His5, Ser9-His10, Phe24-Phe25 and Lys29-Ala30
-
-
-
Oxidized insulin B-chain + H2O
?
-
enzyme primarily hydrolyzes Leu15-Tyr16 bond and secondarily Gln4-His5, Ser9-His10, Phe24-Phe25 and Lys29-Ala30
-
-
-
proCSF + H2O

CSF
-
-
-
-
?
proCSF + H2O
CSF
-
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O

Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
?
Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
?
succinyl-AAPA-p-nitroanilide + H2O

?
-
-
-
?
succinyl-AAPA-p-nitroanilide + H2O
?
-
-
-
-
-
succinyl-AAPF-p-nitroanilide + H2O

succinyl-AAPF + p-nitroaniline
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
-
-
-
?
succinyl-AAPF-p-nitroanilide + H2O
succinyl-AAPF + p-nitroaniline
-
-
-
?
Succinyl-Ala-Ala 4-nitroanilide + H2O

?
-
-
-
-
-
Succinyl-Ala-Ala 4-nitroanilide + H2O
?
-
-
-
-
-
Succinyl-Ala-Ala-Ala 4-nitroanilide + H2O

?
-
-
-
-
-
Succinyl-Ala-Ala-Ala 4-nitroanilide + H2O
?
-
-
-
-
-
succinyl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O

succinyl-AAPL + p-nitroaniline
-
-
-
?
succinyl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O
succinyl-AAPL + p-nitroaniline
-
-
-
?
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O

succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
-
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
-
-
-
-
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
-
-
-
?
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
-
-
-
-
?
Succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide + H2O

?
-
-
-
-
-
Succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide + H2O
?
-
-
-
-
-
ubiquitin + H2O

?
-
from bovine blood cells
-
-
?
ubiquitin + H2O
?
-
from bovine blood cells
-
-
?
Urea-denatured hemoglobin + H2O

?
-
-
-
-
?
Urea-denatured hemoglobin + H2O
?
-
-
-
-
?
Z-Ala-Ala-Leu-p-nitroanilide + H2O

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Z-Ala-Ala-Leu-p-nitroanilide + H2O
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Z-Ala-Ala-Leu-p-nitroanilide + H2O
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additional information

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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
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additional information
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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
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additional information
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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
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additional information
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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
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additional information
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reactions catalyzed: 1. peptide bond hydrolysis, 2. ester bond hydrolysis, 3. transesterification, 4. transpeptidation
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additional information
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reactions catalyzed: 1. peptide bond hydrolysis, 2. ester bond hydrolysis, 3. transesterification, 4. transpeptidation
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additional information
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alteration of substrate specificity by protein engineering
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additional information
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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
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additional information
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substrates bind in a less catalytically favorable conformation after the enzyme has been exposed to organic media for several hours
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additional information
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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
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additional information
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random mutagenesis to enhance the activity of subtilisin in organic solvents
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additional information
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under relatively non-aqueous conditions, immobilized subtilisin is able to synthesize phenylacetic acid ethyl ester
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additional information
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high elastolytic acitivity
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additional information
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preferentially hydrolyzes the ester bond of Ala, but significant hydrolysis is observed with other aliphatic (Gly, Leu) and aromatic (Tyr) amino acids
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additional information
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reactions catalyzed: 1. peptide bond hydrolysis, 2. ester bond hydrolysis, 3. transesterification, 4. transpeptidation
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additional information
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reactions catalyzed: 1. peptide bond hydrolysis, 2. ester bond hydrolysis, 3. transesterification, 4. transpeptidation
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additional information
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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 75°C, 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 40°C
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additional information
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biosynthesis of subtilisin requires participation of an N-terminal prodomain
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additional information
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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 75°C, 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 40°C
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additional information
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high elastolytic acitivity
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additional information
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preferentially hydrolyzes the ester bond of Ala, but significant hydrolysis is observed with other aliphatic (Gly, Leu) and aromatic (Tyr) amino acids
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additional information
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random mutagenesis to enhance the activity of subtilisin in organic solvents
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additional information
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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
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additional information
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reactions catalyzed: 1. peptide bond hydrolysis, 2. ester bond hydrolysis, 3. transesterification, 4. transpeptidation
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additional information
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does not cleave proPhrE
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additional information
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the enzyme shows activity towards complex substrates, e.g. skimmed milk
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additional information
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the enzyme shows activity towards complex substrates, e.g. skimmed milk
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additional information
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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
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additional information
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random mutagenesis to enhance the activity of subtilisin in organic solvents
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additional information
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enzymes prefers cleaving after hydrophobic residues (and in particular P1 leucine)
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additional information
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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
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additional information
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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
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additional information
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displays preference for cleavage after Glu
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additional information
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Sub2 undergoes self-digestion at high concentrations
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additional information
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enzyme is able to hydrolyze a range of fluorogenic substrates, including AFC derivatives of VEID, VAD, YVAD, IETD, VDVAD and LEHD
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additional information
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enzyme is able to hydrolyze a range of fluorogenic substrates, including AFC derivatives of VEID, VAD, YVAD, IETD, VDVAD and LEHD
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additional information
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degrades protein components of both nematode (Meloidogyne sp.) and insect (Phthorimaea opercullella) eggs
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additional information
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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
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additional information
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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
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additional information
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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
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additional information
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autoprocessing of Pro-subtilisin-like serine protease
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(dimethylamino)-1-naphthalenesulfonyl fluoride
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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
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 40%
2,2'-[[(1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphoryl]bis(oxybenzene-4,1-diyl)]diacetic acid
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2,2'-[[(1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphoryl]bis(oxybenzene-4,1-diyl)]diacetic acid
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2,2'-[[(1-[[1-(tert-butoxycarbonyl)-L-prolyl]amino]-3-methylbutyl)phosphoryl]bis(oxybenzene-3,1-diyl)]diacetic acid
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2-phenylethaneboronic acid
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2-phenylethanesulfonic acid
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3,4-dichloroisocoumarin
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4-(2-aminoethyl)-benzenesulfonyl fluoride
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4-(2-aminoethyl)benzenesulfonyl fluoride
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4-(4'-Aminophenylazo)phenylarsonic acid
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4-bromophenacyl bromide
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5-Dimethylaminonaphthalene-1-sulfonate
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circular-dichroism and fluorescence studies of 5-dimethylaminonaphthalene-1-sulfonyl derivative of subtilisin indicate a closely similar structure to that of native subtilisin
alpha2-Macroglobulin
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inhibits alkaline proteolytic activity of purified Arp by 80%
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Benzyloxycarbonyl-(Ala)n-PheCH2Cl
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benzyloxycarbonyl-(Ala)2-PheCH2Cl is the best inhibitor
Benzyloxycarbonyl-Ala-XaaCH2Cl
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Xaa : Gly, Val, Ala, Leu, Phe
Benzyloxycarbonyl-L-phenylalanylbromomethane
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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
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bis(2,3-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(2,3-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(2,5-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(2-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(2-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(3,4,5-trimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(3,4,5-trimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(3,4-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(3,4-dimethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(3-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(3-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(3-methoxyphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(4-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(4-chlorophenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(4-ethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(4-ethylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(4-methoxyphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(4-methylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis(4-tert-butylphenyl) (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis(4-tert-butylphenyl) (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis[4-(methylsulfonyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis[4-(propan-2-yl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis[4-(propan-2-yl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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bis[4-(sulfanylmethyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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bis[4-(sulfanylmethyl)phenyl] (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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Brij 35
B6VFQ8;
at 37°C 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
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subtilisin BPN'
Ca2+
B6VFQ8;
at 37°C and pH of 7.5, 1 mM reduces prosubtilisin JB1 relative activity to 57% and 5 mM reduces prosubtilisin JB1 relative activity to 45%
Chiral amine- and aminoalcohol-derivatives
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Chymotrypsin I inhibitor from potato
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subtilisin Carlsberg, subtilisin BPN'
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chymotrypsin inhibitor 2
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chymotrypsin inhibitor 2 mutant M59A
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chymotrypsin inhibitor 2 mutant M59F
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chymotrypsin inhibitor 2 mutant M59G
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chymotrypsin inhibitor 2 mutant M59K
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chymotrypsin inhibitor 2 mutant M59Y
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chymotrypsin inhibitor 2 mutant Y61A
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Co2+
B6VFQ8;
at 37°C 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
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Kazal-type inhibitor from the hepatopancreas of the Carcinoscorpius rotundicauda, potently inhibits subtilisin
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diisopropyl fluorophosphate
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diisopropylphosphofluoridate
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Dipeptidyl chloromethyl ketones
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diphenyl (1-[[(benzyloxy)carbonyl]amino]-2-phenylethyl)phosphonate
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diphenyl (1-[[(benzyloxy)carbonyl]amino]-3-methylbutyl)phosphonate
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E-64
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 31%
E-64c
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 25%
EGTA
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 34%
EPI1a
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four-cysteine atypical Kazal-domain of protease inhibitor EPI1 from Phytophthora infestans. 80% inhibition at 0.00015 mM
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EPI1b
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typical Kazal-domain of protease inhibitor EPI1 from Phytophthora infestans, little inhibitory effect
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fungal protease inhibitor F
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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
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guanidine hydrochloride
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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
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Hg+
B6VFQ8;
at 37°C 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
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noncompetitive inhibition
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human proteinase inhibitor 9
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PI9, serine proteinase inhibitor
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Inhibitor from Dolichos biflorus
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purification and properties
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Inhibitor from egg white
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Inhibitor from seeds of Canavalia lineata
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Inhibitor from seeds of Setaria italica
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purification and characterization
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Inhibitor from Streptomyces sp.
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Inhibitor from Streptomyces virginiae
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primary structure
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Inhibitor from Vigna unguiculata subsp. cylindrica
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Inhibitor of Amaranthus caudatus seeds
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Inhibitor of trypsin from soybean
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K+
B6VFQ8;
at 37°C 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
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leupeptin
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inhibits the enzymatic activity by 20%
Mg2+
B6VFQ8;
at 37°C and pH of 7.5, 1 mM reduces prosubtilisin JB1 relative activity to 79% and 5 mM reduces prosubtilisin JB1 relative activity to 48%
N-((tert-Butoxycarbonyl)alanylprolylphenylalanyl)-O-benzoylhydroxylamine
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N-(tert-butoxycarbonyl)-L-valyl-N-(1-[bis[4-(sulfanylmethyl)phenoxy]phosphoryl]-2-phenylethyl)-L-prolinamide
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N-(tert-butoxycarbonyl)-L-valyl-N-[1-(diphenoxyphosphoryl)-2-phenylethyl]-L-prolinamide
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N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(3,4,5-trimethylphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
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N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(4-methoxyphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
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N-(tert-butoxycarbonyl)-L-valyl-N-[1-[bis(4-tert-butylphenoxy)phosphoryl]-3-methylbutyl]-L-prolinamide
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N-acetyl-L-tryptophan amide
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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
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product inhibition
N-benzyloxycarbonyl-Ala-Pro-Phe-chloromethyl ketone
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synthetic inhibitor
N-ethylmaleimide
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 15%
N-tosyl-L-phenylalanyl chloromethyl ketone
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N1-18[ISP]
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synthetic peptide corresponding to the N-terminal extension behaves as a mixed noncompetitive inhibitor of active ISP
p-chloromercuribenzoate
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partial inhibition
p-Nitrophenylarsonate
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Pepstatin
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almost completely inhibits acid proteolytic activity, does not inhibit alkaline proteolytic activity of purified Arp
pepstatin A
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 48%
Peptidyl chloromethyl ketones
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phenylmethanesulfonyl fluoride
phenylmethylsulfonyl fluoride
Phenylmethylsulphonyl fluoride
-
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Potato extract
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subtilisin BPN'
-
propeptide
-
inhibition in a concentration-dependent manner. Wild-type propeptide is more potent than G56W-, G56S- and G56E-propeptide
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Suc-Val-Pro-PheP(OPh)2
-
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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 proteinase–inhibitor complexes. The domains of TI-II appear to bind the proteinase independently of each other
-
tosyl-Phe chloromethyl ketone
Triton X-100
B6VFQ8;
at 37°C 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
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Tween 20
B6VFQ8;
at 37°C and pH of 7.5, 0.01% reduces prosubtilisin JB1 relative activity to 53% and 0.05% reduces prosubtilisin JB1 relative activity to 70%
1,4-dioxane

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104fold decrease in the enzyme’s 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
antipain

-
-
antipain
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 75%
antipain
-
totally inhibits recombinant Sub2
Aprotinin

-
-
Aprotinin
-
inhibits the enzymatic activity by 15%
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
-
chymostatin

-
-
chymostatin
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 95%
chymostatin
-
totally inhibits recombinant Sub2
Cu2+

-
strain DJ-4, strong inhibition
Cu2+
B6VFQ8;
at 37°C 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%
EDTA

5 mM partially inhibits by 35%, in 100 mM Tris-HCl buffer, pH 7.0 at 15°C for 20 min
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
EDTA
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 52%
EDTA
-
subtilisin exhibits little activity at 80°C in the presence of 10 mM. Subtilisin-like serine protease is fully active even in the presence of 10 mM EDTA
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 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 Amaranthus caudatus seeds
-
subtilisin Novo (BPN') from Bacillus amyloliquefaciens, subtilisin Carlsberg from Bacillus licheniformis
-
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
phenylmethanesulfonyl fluoride

-
-
phenylmethanesulfonyl fluoride
-
-
phenylmethanesulfonyl fluoride
-
totally inhibits recombinant Sub2
phenylmethylsulfonyl fluoride

1 mM completely inhibits, in 100 mM Tris-HCl buffer, pH 7.0 at 15°C for 20 min
phenylmethylsulfonyl fluoride
-
1 mM, 82% inhibition
phenylmethylsulfonyl fluoride
-
-
phenylmethylsulfonyl fluoride
-
strain DJ-4
phenylmethylsulfonyl fluoride
-
-
phenylmethylsulfonyl fluoride
-
1 mM, 5% residual activity
phenylmethylsulfonyl fluoride
B6VFQ8;
at 37°C and pH of 7.5, 0.1 mM inhibits prosubtilisin JB1 by 79%
phenylmethylsulfonyl fluoride
-
inhibits alkaline proteolytic activity of purified Arp by 100%, does not inhibit acid proteolytic activity
PMSF

-
-
PMSF
-
circular-dichroism and fluorescence studies of PMSF derivative of subtilisin indicate a closely similar structure to that of native subtilisin
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 subtilisin–sodium 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
B6VFQ8;
at 37°C and pH of 7.5, 0.01% reduces prosubtilisin JB1 relative activity to 52% and 0.05% reduces prosubtilisin JB1 relative activity to 29%
tosyl-Phe chloromethyl ketone

-
not
tosyl-Phe chloromethyl ketone
-
-
turkey egg white inhibitor

-
-
-
turkey egg white inhibitor
-
-
-
Zn2+

-
strain DJ-4, strong inhibition
Zn2+
-
1 mM, 59% residual activity
Zn2+
B6VFQ8;
at 37°C 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
additional information

-
enzyme-specific, thermostable inhibitor from Physarum polycephalum with molecular mass of 32-33 kDa, 50% inhibition at 0.00014 mM
-
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
-
not inhibitory: Tween-20, Tween-40, Tween-60, Tween-80 at 1%, sodium dodecylsulfate at 0.2%, for 1 h at 30°C. Not inhibitory: iodoacetate, ethylacetimide, phenylglyoxal, N-ethylmaleimide, N-bromosuccinimide
-
additional information
-
molecular dynamics simulations, residue displacement correlations, and inhibitor design based on the enzyme inhibitor comlpex of TI-II and subtilisin, overview
-
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: EDTA at 5 mM
-
additional information
-
not inhibitory: sodium dodecylsulfate at 1 mM
-
additional information
B6VFQ8;
at 37°C 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
-
not inhibited by 1,10 phenanthroline
-
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
-
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
-
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