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Information on EC 3.4.24.27 - thermolysin and Organism(s) Bacillus thermoproteolyticus and UniProt Accession P00800
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3.4.24.27 thermolysinSpecify your search results
Word Map
- 3.4.24.27
- chymotrypsin
- elastase
- metalloprotease
- subtilisin
- staphylococcus
-
edman
- pepsin
- aureus
- carboxypeptidase
- endopeptidase
- bromide
-
cyanogen
- collagenase
- proteinases
- dipeptide
- angiotensin
- pronase
- metalloproteinases
- alpha-chymotrypsin
-
thermolytic
- hydrolysates
- metalloendopeptidase
- stearothermophilus
-
endoproteinase
- phosphoramidon
-
i-converting
- enkephalinase
- rhodopsin
-
2-macroglobulin
-
3.4.24.11
-
cell-binding
- alcalase
-
ace-inhibitory
-
hexxh
-
dispase
- aspartame
- thiorphan
- neprilysin
-
s-carboxymethylated
-
half-cystine
- astacin
- synthesis
- industry
- nutrition
- food industry
- diagnostics
- medicine
- analysis
The taxonomic range for the selected organisms is: Bacillus thermoproteolyticus
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
thermolysin, thermolysin-like protease, tlp-ste, thermostable neutral proteinase, lic13322, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Bacillus thermoproteolyticus neutral proteinase
-
-
-
-
Proteinase, Bacillus thermoproteolyticus neutral
-
-
-
-
Thermoase
-
-
-
-
Thermoase Y10
-
-
-
-
Thermostable neutral proteinase
-
-
-
-
TLN
TLN
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
preferential cleavage: -/-Leu > -/-Phe
classical molecular dynamics simulation., and ab initio quantum mechanics/molecular mechanics, QM/MM, investigation of the Glu143-assisted water addition mechanism in thermolysin peptide hydrolysis. The mechanism consists of three distinct steps: (i) a Zn-bound water molecule is deprotonated by Glu143 and attacks the carbonyl bond of the substrate; (ii) Glu143 transfers the proton to the amide nitrogen atom; (iii) the nitrogen atom is protonated and the peptide bond is irreversibly broken. Transition state stabilization for nucleophilic attack is achieved by formation of a weak coordination bond between the substrate carbonyl oxygen atom and the Zn ion and of three strong hydrogen bonds between the substrate and protonated His231 and two solvent molecules, overview
-
preferential cleavage: -/-Leu > -/-Phe
substrate turnover is associated with hinge bending that leads to a closed conformation. Product release regenerates the open form, such that steady-state catalysis involves a continuous closing/opening cycle. Elements in the periphery of the two lobes are most mobile. These peripheral regions undergo quite pronounced structural changes during the catalytic cycle. In contrast, active site residues exhibit only a moderate degree of backbone flexibility, and the central zinc appears to be in a fairly rigid environment. The hydrogen/deuterium exchange behavior of catalytically active thermolysin is indistinguishable from that of the free enzyme
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9073-78-3
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2-N-(4-[4'-N',N'-(dimethylamino)phenylazo]-benzoyl-L-serinyl-L-phenylalanylamido)-N''-ethylaminonaphthalene-5-sulfonic acid + H2O
?
-
-
-
?
carbobenzoxy-L-aspartic acid + L-phenylalanine methyl ester
carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
condensation, the enzyme is enantioselective for the desired L-phenylalanine methyl ester substrate from a racemic mixture of DL-phenylalanine methyl ester. In contrast, although both enantiomers of carbobenzoxy-L-aspartic acid can bind to the enzyme, only carbobenzoxy-L-aspartic acid is used since carbobenzoxy-D-aspartic acid inhibits the enzyme, substrate carbobenzoxy-L-aspartic acid binding structures, detailed overview
precipitation as the water-insoluble Phe-OMe salt drives the overall reaction in the direction of peptide synthesis
-
?
cellular prion protein + H2O
?
thermolysin degrades cellular prion protein while preserving both proteinase K-sensitive and proteinase K-resistant isoforms of disease-related prion protein in both rodent and human prion strains. In variant Creutzfeldt-Jakob disease, up to 90% of total prion protein present in the brain resists degradation with thermolysin, whereas only about 15% of this material resists digestion by proteinase K
-
-
?
N-carbobenzoxy-L-Asp-L-Phe-methyl ester + H2O
N-carbobenzoxy-L-Asp + L-Phe-methyl ester
-
-
-
?
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester + H2O
?
-
-
-
?
(7-methoxycoumarin-4-yl) acetyl-L-Pro-L-Leu-Gly-L-Leu-[N3-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl]-L-Ala-L-Arg-NH2 + H2O
?
(7-methoxycoumarin-4-yl)acetyl-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys-(2,4-dinitrophenyl)-OH + H2O
?
-
a bradykinin-like substrate
-
-
?
(7-methoxycoumarin-4yl) acetyl-L-Pro-L-Leu-Gly-L-Leu-[N3-(2,4-dinitrophenyl)-L-2,3-diamino-propionyl]-L-Ala-L-Arg-NH2 + H2O
?
-
-
-
-
?
(europium(III) complex of a modified terpyridine)-K1K1K1-GFSAK1K1K-black hole quencher 2 + H2O
?
-
thermolysin cleaves the substrates at the glycine-phenylalanine bond
-
-
?
(europium(III) complex of a modified terpyridine)-K1K2K2GFSAK2K-black hole quencher 2 + H2O
?
-
thermolysin cleaves the substrates at the glycine-phenylalanine bond
-
-
?
(europium(III) complex of a modified terpyridine)-K1K2K2GFSAK2K2K-black hole quencher 2 + H2O
?
-
thermolysin cleaves the substrates at the glycine-phenylalanine bond
-
-
?
2-hydroxy-N-(4-methyl-2-nitrophenyl)-3-nitrobenzamide + H2O
?
-
-
-
-
?
3-(2-furylacryloyl)-glycyl-L-leucine amide + H2O
?
-
-
-
?
5-bromo-N-(4-bromophenyl)-2-hydroxy-3-nitro-benzamide + H2O
?
-
-
-
-
?
Ac-Gly-Leu-Ala-methylamide + H2O
?
-
model substrate, enzyme-substrate complex, docking structures, overview
-
-
ir
alphaS1-casein + H2O
caseicin A + ?
-
-
antimicrobial peptide product caseicin A = IKHQGLPQE
-
?
bovine alpha-lactalbumin + H2O
?
-
reaction at 25°C and 70°C under nonreducing conditions. At 25°C, substrate undergoes limited hydrolysis leading to peptides no longer degraded. At 70°C, protein is first quickly cleaved, then unfolded, leading to the release of intermediate peptides that may be further degraded
-
-
?
bovine beta-lactoglobulin A + H2O
?
-
analysis of 25 peptides released by enzyme at 37°C, comparison with peptides relased at 25°C, 60 and 80°C. Test of peptides for angiotensin-converting enzyme inhibiting activity
-
-
?
casein + H2O
?
-
-
-
-
?
furylacryloyl-Gly-Leu-NH2 + H2O
furylacryloyl-Gly + Leu-NH2
-
-
-
-
?
H-Gly-Phe-Ser-Ala-Lys-Asn-Gln-Ser-Asn-Gln-Arg-OH + H2O
?
-
-
-
?
Mca-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(Dnp)-OH + H2O
?
-
-
-
-
?
N,N'-diBoc-dityrosyl-(Ile-isoniazid)2 + H2O
N,N'-diBoc-dityrosyl + 2 Ile-isoniazid
-
-
-
-
?
N,N'-diBoc-dityrosyl-(Phe-isoniazid)2 + H2O
N,N'-diBoc-dityrosyl + 2 Phe-isoniazid
-
-
-
-
?
N-(2,3-dimethylphenyl)-2-hydroxy-3-nitro-benzamide + H2O
?
-
-
-
-
?
N-(2,4-dimethylphenyl)-2-hydroxy-3-nitro-benzamide + H2O
?
-
-
-
-
?
N-(2,5-dimethylphenyl)-2-hydroxy-3-nitrobenzamide + H2O
?
-
-
-
-
?
N-(2-chloro-4-nitrophenyl)-2-hydroxy-3-nitro-benzamide + H2O
?
-
-
-
-
?
N-(2-chloro-6-methylphenyl)-2-hydroxy-3-nitrobenzamide + H2O
?
-
-
-
-
?
N-(5-chloro-2-methoxyphenyl)-2-hydroxy-3-nitro-benzamide + H2O
?
-
-
-
-
?
N-(Benzyloxycarbonyl)-L-Phe + L-Phe methyl ester
N-(Benzyloxycarbonyl)-L-Phe-L-Phe methyl ester + H2O
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester + H2O
N-benzyloxycarbonyl-L-Asp + L-Phe-methyl ester
-
-
-
-
?
N-carbobenzoxy-L-Asp-L-Phe methyl ester + H2O
?
-
-
-
-
?
N-carbobenzoxy-L-Asp-L-Phe-methyl ester + H2O
?
-
-
-
-
?
N-carbobenzoxy-L-Asp-L-Phe-methyl ester + H2O
N-carbobenzoxy-L-Asp + L-Phe-methyl ester
-
-
-
-
?
N-carbobenzoxy-L-Asp-L-Phe-methyl ester + H2O
N-carbobenzoxy-L-aspartic acid + L-phenylalanine methyl ester
-
-
-
-
r
N-carboxybenzoyl-L-aspartyl-L-phenylalanine methyl ester + H2O
?
-
-
-
?
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide + H2O
?
-
-
-
-
?
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O
N-[3-(2-furyl)acryloyl]-glycine + L-leucine amide
-
-
-
-
?
N-[3-(2-furyl)acryloyl]-L-leucine-L-alanine amide + H2O
?
-
i.e. FALAA
-
-
?
Nalpha-benzyloxycarbonyl-L-aspartyl-L-phenylalanine methyl ester + H2O
?
-
-
-
?
Pro-urokinase + H2O
?
-
thermolysin activates thrombin-inactivated pro-urokinase nearly as rapidly as it does the native zymogen, cleavage of Arg156-Phe157 and Lys158-Ile159
-
-
?
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O
?
-
-
-
?
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O
?
i.e. FAGLA
-
-
?
(7-methoxycoumarin-4-yl) acetyl-L-Pro-L-Leu-Gly-L-Leu-[N3-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl]-L-Ala-L-Arg-NH2 + H2O
?
-
fluorescent substrate
-
?
(7-methoxycoumarin-4-yl) acetyl-L-Pro-L-Leu-Gly-L-Leu-[N3-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl]-L-Ala-L-Arg-NH2 + H2O
?
-
i.e. MOCAc-PLGL(Dpa)AR, fluorescent substrate
-
-
?
benzyloxycarbonyl-Asp + Phe-methylester
benzyloxycarbonyl-Asp-Phe methyl ester + H2O
-
-
-
?
benzyloxycarbonyl-Asp + Phe-methylester
benzyloxycarbonyl-Asp-Phe methyl ester + H2O
-
-
-
?
Collagen + H2O
?
-
collagen in solid articular cartilage at 70°C. Overnight digestion with thermolysin completely solubilized cartilage. Following thermolysin treatments, almost all glycosaminoglycans are extracted from the cartilage
-
-
?
N-(Benzyloxycarbonyl)-L-Phe + L-Phe methyl ester
N-(Benzyloxycarbonyl)-L-Phe-L-Phe methyl ester + H2O
-
-
-
?
N-(Benzyloxycarbonyl)-L-Phe + L-Phe methyl ester
N-(Benzyloxycarbonyl)-L-Phe-L-Phe methyl ester + H2O
-
-
-
?
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O
?
-
-
-
r
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O
?
-
i.e. FAGLA
-
-
?
additional information
?
-
binding of substrates to the active site of thermolysin, overview
-
-
?
additional information
?
-
-
specificity overview: various synthetic peptides
-
-
?
additional information
?
-
-
specificity overview: oligopeptides
-
-
?
additional information
?
-
-
enzyme additionally catalyzes the transesterification of vinyl laurate to several sucrose-containing tri- and tetrasaccharides. Preferred position of acylation is the 2-OH group of the alpha-D-glucopyranose moiety linked 1 to 2 to the beta-D-fructofuranose unit
-
-
?
additional information
?
-
-
enzyme catalyzes the formation of beta-cyclodextrin esters using vinyl esters of butyrate, decanoate and laurate, as acyl donors in dimethylsulfoxide. Esterification occurs exclusively at the glucose C-2 position. Enzyme also catalyzes the synthesis of alpha-, beta-, gamma- and maltosyl-beta-cyclodextrin esters with vinyl laurate as the acyl donor in dimethylsulfoxide and dimethylformamide
-
-
?
additional information
?
-
-
enzyme prefers basic resiudes in P3 position
-
-
?
additional information
?
-
-
digestion of brain homogenates from healthy and scrapie-affected sheep as well as healthy and BSE-affected cattle, the PK digestion results in the amino-terminal truncation of PrPSc-producing PrP species with molecular weights of approx 17, 21, and 27 kDa that can only be detected with antibodies binding to the carboxy-terminal region of PrP, the so-called protease-resistant core or PrP27-30, overview
-
-
?
additional information
?
-
-
thermolysin is a thermostable neutral metalloproteinase and performs autocatalytic cleavage for pro-enzyme activation
-
-
?
additional information
?
-
-
thermolysin performs Co2+-stimulable autolysis
-
-
?
additional information
?
-
-
the major site for thermolysin cleavage specificity, (the S1' site), accepts large hydrophobic residues. Thermolysin preferentially cleaves at the N-terminal side of hydrophobic or bulky amino side chains such as Leu, Phe, Ile and Val. Thermolysin also cleaves bonds of Met, His, Tyr, Ala, Asn, Ser, Thr, Gly, Lys, Glu or Asp at the P1' site
-
-
?
additional information
?
-
-
isolated intact chloroplast from Spinacia oleracea are treated with thermolysin, mass spectrometric analysis and two-dimensional analysis of shedded envelope proteins, including 28 kDa ribonucleoprotein, cytosolic HSP70/Com70, translocon Tic40-like protein, ClpC, HSP70, and hexokinase 1, overview
-
-
?
additional information
?
-
-
synthesis and evaluation of substrates useful for the selective and sensitive assay of thermolysin, overview
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
isolated intact chloroplast from Spinacia oleracea are treated with thermolysin, mass spectrometric analysis and two-dimensional analysis of shedded envelope proteins, including 28 kDa ribonucleoprotein, cytosolic HSP70/Com70, translocon Tic40-like protein, ClpC, HSP70, and hexokinase 1, overview
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
NaCl
activates the mutant enzymes at 4 M to 17-19fold of wild-type enzyme activity, overview
Zn2+
Ca2+
Co2+
K+
-
activates, preference of monovalent cations in descending order: Na+, K+, Li+
Li+
-
activates, preference of monovalent cations in descending order: Na+, K+, Li+
Na+
NaCl
Zinc
Zn2+
additional information
Zn2+
comparison of metal preferences of Escherichia coli peptide deformylase and Bacillus thermoproteolyticus thermolysin. Both enzymes catalyze via the same chemical steps, and reproduce their different preferences for zinc or iron as competent cofactors. In thermolysin, the substrate is strongly activated and can serve as the fifth coordination ligand of zinc prior to the chemical steps. When iron replaces zinc, its stronger interaction with the hydroxide ligand may lead to higher activation barrier in thermolysin
Ca2+
-
there are two adjacent calcium ions seemingly firmly bound inside the surface of the molecule by chelation to five acidic groups: Asp138, Glu177, Asp185, Glu190 and Asp191, two additional calcium binding sites are at exposed surface regions, one chelated by Asp57 and possibly also by Asp59 and the other chelated by Asp200
Ca2+
-
four Ca2+ per enzyme molecule are required for enzyme stability
Ca2+
-
four ions per enzyme molecule required for stability
Ca2+
-
thermolysin consists of four Ca2+ ligand ions necessary for stability
Ca2+
-
thermolysin has four calcium ions responsible for its thermostability
Ca2+
-
CaCl2 does not exert any significant influence on the activity of immobilized thermolysin, but stabilizes the enzyme in absence of Zn2+
Co2+
-
active zinc ion in thermolysin can be substituted by Co2+, doubles activity
Co2+
-
increases the activity 3-4fold at up to 2 mM, but inhibits at 2-18 mM, activation-and-inhibition dual effects of Co2+ ion are analysed kinetically, Co2+-dependent activation is inhibited competitively by Zn2+ ion at 0.0001-0.001 mM
Co2+
-
the active site zinc atom of thermolysin is replaced by cobalt
Na+
-
activates, preference of monovalent cations in descending order: Na+, K+, Li+, the bell-shaped pH dependence profile of the FAGLA-hydrolyzing activity of thermolysin is shifted from pH 5.4 to pH 6.7 by the addition of 4 M NaCl
Na+
-
4 M Na+ stimulates the hydrolytic activity of wild type thermolysin about 13fold
NaCl
-
induces enzyme activation, activation of mutant enzymes is reduced compared to the wild-type enzyme
Zinc
-
the three zinc ligands are two histidines and glutamic acid
Zn2+
-
a zinc metalloproteinase that contains a HEXXH motif, one Zn2+ per enzyme molecule is required for activity
Zn2+
-
one ion per enzyme molecule essential for activity, the enzyme contains the HEXXH motif constituting the zinc catalytic site
Zn2+
-
one ion per enzyme molecule required for activity
Zn2+
-
stereochemical relationships between Gln128, Glu143, Gln225, Asp226, His231 and active site Zn2+ of thermolysin, overview
Zn2+
-
zinc metalloproteinase, one ion per enzyme molecule required for activity
Zn2+
-
residues H142, H146, and E166 coordinate the catalytic zinc
Zn2+
-
a single catalytic Zn2+ ion is essential for hydrolytic activity
Zn2+
-
thermolysin consists of three Zn2+ ligand ions necessary for activity
Zn2+
-
the enzyme has a catalytic zinc ion at the active site cleft with a tetrahedral coordination formed by the two histidines of a HEXXH motif, and a glutamic acid located 18-72 residues C-terminal of the HEXXH motif. The fourth zinc coordinating ligand in the free enzyme is a water molecule
additional information
-
thermolysin activity as well as its stability is remarkably enhanced by high concentration of neutral salts consisting of Na+, K+, Cl-, and Br- in the synthesis and hydrolysis of N-carbobenzoxy-L-aspertyl-L-phenylalanine methyl ester and hydrolysis of N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
carbobenzoxy-L-phenylalanine-phosphonamidate-L-leucyl-L-alanine
a potent phosphonamidate transition state analogue inhibitor
(E)-N-(naphthalen-1-yl)-N'-(4-oxo-2-phenylquinazolin-3(4H)-yl)acetimidamide
-
low inhibitory activity
2-(4-chlorophenyl) quinazolin-4(3H)-one
-
low inhibitory activity
2-(4-methylphenyl) quinazolin-4(3H)-one
-
low inhibitory activity
2-(4-methylphenyl)-3-(1,3-thiazol-2-yl) quinazolin-4(3H)-one
-
-
2-(4-oxo-2-methylquinazolin-3(4H)-yl) guanidine
-
low inhibitory activity
2-(N-Bromoacetyl-N-hydroxyamino)-4-methylpentanonitrile
-
irreversible
2-benzamido-N-(3-(4-oxo-2-phenylquinazolin-3(4H)-yl)propyl)benzamide
-
low inhibitory activity
2-chloro-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
-
low inhibitory activity
2-hydroxy-N-(4-methyl-2-nitrophenyl)-3-nitrobenzamide
-
competitive
2-phenyl-3-(1, 3-thiazol-2-yl) quinazolin-4(3H)-one
-
low inhibitory activity
2-phenyl-3-[[(1E)-phenylmethylene] amino]-2,3-dihydroquinazolin-4(1H)-one
-
low inhibitory activity
2-[(biphenyl-4-ylsulfonyl)[2-(hydroxyamino)-2-oxoethyl]amino]-N-[2-(4-sulfamoylphenyl)ethyl]acetamide (non-preferred name)
-
-
2-[benzyl[2-(hydroxyamino)-2-oxoethyl]amino]-N-[2-(4-sulfamoylphenyl)ethyl]acetamide (non-preferred name)
-
the phenyl group of the strong binder occupies the S'2-subpocket, while a second ring system occupy the S1-subpocket in both thermolysin and pseudolysin, EC 3.4.24.27
2-[benzyl[2-(hydroxyamino)-2-oxoethyl]amino]-N-[3-(4-phenylpiperazin-1-yl)propyl]acetamide (non-preferred name)
-
the phenyl group of the strong binder occupies the S'2-subpocket, while a second ring system occupy the S1-subpocket in both thermolysin and pseudolysin, EC 3.4.24.26
2-[[2-(hydroxyamino)-2-oxoethyl][(4-methoxyphenyl)sulfonyl]amino]-N-[2-(4-sulfamoylphenyl)ethyl]acetamide (non-preferred name)
-
-
2-[[2-(hydroxyamino)-2-oxoethyl][(4-phenoxyphenyl)sulfonyl]amino]-N-[2-(4-sulfamoylphenyl)ethyl]acetamide (non-preferred name)
-
-
3-(2-aminoethyl)-2-(4-methylphenyl) quinazolin-4(3H)-one
-
low inhibitory activity
3-(2-hydroxyethyl) quinazolin-4(3H)-one
-
low inhibitory activity
3-(3-aminopropyl)-2-(4-methylphenyl) quinazolin-4(3H)-one
-
low inhibitory activity
3-(4-iodophenyl)-2-phenylquinazolin-4(3H)-one
-
low inhibitory activity
3-(isopropylideneamino)-2,2-dimethyl-2,3-dihydroquinazolin-4(1H)-one
-
potent inhibitor
3-([(1E)-[4-(dimethylamino) phenyl]methylene]amino)-2-methylquinazolin-4(3H)-one
-
-
3-([(1E)-[4-(dimethylamino) phenyl]methylene]amino)-2-phenylquinazolin-4(3H)-one
-
-
3-amino-2-(hydrazinomethyl) quinazolin-4(3H)-one
-
low inhibitory activity
3-phenyl-2-(trifluoromethyl) quinazolin-4(3H)-one
-
potent inhibitor
3-Phenylpropionyl-L-Phe
-
crystallographic study of the binding to thermolysin
3-[[(1E)-(2-hydroxynaphthalen-1-yl)methylene]amino]-2-phenylquinazolin-4(3H)-one
-
low inhibitory activity
3-[[(1E)-(3-chlorophenyl)methylene]amino]-2-phenylquinazolin-4(3H)-one
-
-
3-[[(1E)-(4-fluorophenyl)methylene]amino]-2-phenylquinazolin-4(3H)-one
-
low inhibitory activity
4-methyl-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)benzamide
-
low inhibitory activity
5-bromo-N-(4-bromophenyl)-2-hydroxy-3-nitro-benzamide
-
competitive
carbobenzoxy-L-Phe
-
crystallographic study of the binding to thermolysin
ClCH2CO-DL-(N-OH)Leu-OCH3
-
specific, irreversible, pH-dependence of inhibition
Cu2+-Cys-Gly-His-Lys
-
stimulation at concentration up to 0.01 mM, inhibition at higher concentrations of 0.01-0.1 mM, binding and kinetics, overview
ethanimidic acid N-[4-oxo-2-phenyl-3(4H)-quinazolinyl]-ethyl ester
-
-
ethyl (4-oxo-3,4-dihydroquinazolin-2-yl)acetate
-
low inhibitory activity
Hydroxamic acid inhibitors
-
binding to thermolysin suggests a pentacoordinate zinc intermediate
-
methyl 2-[(trifluoroacetyl) amino] benzoate
-
low inhibitory activity
N-(2,3-dimethylphenyl)-2-hydroxy-3-nitro-benzamide
-
competitive
N-(2,4-dimethylphenyl)-2-hydroxy-3-nitro-benzamide
-
competitive
N-(2-chloro-4-nitrophenyl)-2-hydroxy-3-nitro-benzamide
-
competitive
N-(2-chloro-6-methylphenyl)-2-hydroxy-3-nitrobenzamide
-
competitive
N-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
-
low inhibitory activity
N-(2-methyl-4-oxoquinazolin-3(4H)-yl)benzamide
-
low inhibitory activity
N-(2-phenyl-4-oxoquinazolin-3(4H)-yl)acetamide
-
low inhibitory activity
N-(5-chloro-2-methoxyphenyl)-2-hydroxy-3-nitro-benzamide
-
competitive
N-chloroacetyl-N-hydroxyleucine methyl ester
-
irreversible inhibition
N-chloroacetyl-N-hydroxyleucyl-alanyl-glycinamide
-
irreversible inhibition
n-Pentanol
-
saturation concentration of activation at 60%, inhibition at higher concentration
N-[(2R)-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl]-N-[(4-phenoxyphenyl)sulfonyl]glycine
-
-
Z-L-phenylalanine
-
enzyme binding structure and kinetics, chemical shift of the carboxylate carbon upon enzyme binding, overview. The carbobenzyloxyl protecting group and not the phenylalanyl phenyl group that is bound in the S1' specificity pocket and the alpha carboxylate group is directly coordinated to the active site zinc atom
Z-L-tryptophan
-
inhibits full length stromelysin_1-477 and truncated stromelysin_100-264, enzyme binding structure and kinetics, chemical shift of the carboxylate carbon upon enzym ebinding, overview. The tryptophan side chain can bind in the S1 specificity site of stromelysin with the tryptophan alpha carboxylate group coordinated to the active site zinc atom. L-tryptophan binds equally strongly to zinc or cobalt substituted thermolysin
[(biphenyl-4-ylmethyl)[2-(hydroxyamino)-2-oxoethyl]amino]acetic acid
-
-
[(biphenyl-4-ylsulfonyl)[2-(hydroxyamino)-2-oxoethyl]amino]acetic acid
-
-
[1-[2-(hydroxyamino)-2-oxoethyl]-2-[3-(4-phenylpiperazin-1-yl)propyl]hydrazinyl]acetic acid
-
-
[[(4-methoxyphenyl)sulfonyl](2-oxo-2-[[2-(4-sulfamoylphenyl)ethyl]amino]ethyl)amino]acetic acid
-
-
[[2-(hydroxyamino)-2-oxoethyl](4-nitrobenzyl)amino]acetic acid
-
-
[[2-(hydroxyamino)-2-oxoethyl](4-phenoxybenzyl)amino]acetic acid
-
-
[[2-(hydroxyamino)-2-oxoethyl][(4-methoxyphenyl)sulfonyl]amino]acetic acid
-
-
[[2-(hydroxyamino)-2-oxoethyl][(4-phenoxyphenyl)sulfonyl]amino]acetic acid
-
-
Co2+
-
competitive, 72% inhibition at 18 mM, partial protection or reverse effect by Ca2+ at 0.5 mM, Co2+ show partly Ca2+-sensitive and partly Ca2+-insensitive inhibition, molecular mechanism, Co21 accelerates the autolysis, overview
Co2+
-
increases the activity 3-4fold at up to 2 mM, but inhibits at 2-18 mM, activation-and-inhibition dual effects of Co2+ ion are analysed kinetically
dipeptides
-
crystallographic study of the binding to thermolysin
N-Phosphoryl-L-Leu amide
-
thermolysin-inhibitor complexes examined by NMR spectroscopy
phosphoramidon
-
the OH-group is not essential for the binding to thermolysin
phosphoramidon
-
i.e. N-(alpha-L-rhamnopyranosyloxyphospho)-L-Leu-L-Trp, thermolysin-inhibitor complexes examined by NMR spectroscopy
phosphoramidon
-
crystallographic study of the complex of phosphoramidon with thermolysin
additional information
not inhibited by 3-hydroxy-2-methylthieno[3,2-d]pyrimidine-4(3H)-one and 3-(acetylamino)-4-methylthiophene-2-carboxylic acid
-
additional information
binding of inhibitors to the active site of thermolysin, structure overview
-
additional information
-
identification of inhibitors by structure based virtual screening and study of binding modes by docking
-
additional information
-
not inhibited by 2-phenyl-3-[(E)-phenyldiazenyl]-1,2,3,4-tetrahydro-10H[1,2,4]triazino[6,1b]-quinazolin-10-one, 3-mesityl-2-phenylquinazolin-4(3H)-one, 3-(2-hydroxyethyl)-2-methylquinazolin-4(3H)-one, and 3-amino-2-phenylquinazolin-4(3H)-one
-
additional information
-
inhibitor synthesis, docking analysis and binding structure, molecular modeling, overview. When the compounds possess two ring systems, the largest and most electron rich ring system seems to occupy the S1-subpocket. The fourth zinc coordinating ligand in the free enzyme is a water molecule. Upon inhibitor binding this water molecule is replaced by a metal binding group of the inhibitor
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ca2+
-
accelerated hydrolysis of some tryptic peptides derived from bovine beta-casein in presence of Ca2+, while other peptides are not affected
Cu2+-Cys-Gly-His-Lys
-
stimulation at concentration up to 0.01 mM, inhibition at higher concentrations of 0.01-0.1 mM, binding and kinetics, overview
n-Pentanol
-
saturation concentration of activation at 60%, inhibition at higher concentration
NaCl
-
up to 40fold increase in activity in presence of 4 M NaCl, substrate MOCAc-PLGL(Dpa)AR. Degree of activation depends on substrate
additional information
-
direct effect of organic solvents on the microenvironment of the enzyme largely depends on the molecular structure of the solvent
-
additional information
-
no change in hydrolysis of tryptic peptides derived from beta-casein by presence of Na+
-
additional information
-
microwave power output increases the level of activity of the immobilized thermolysin by 4.5fold, optimally at 40 W, overview
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.002
(7-methoxycoumarin-4yl) acetyl-L-Pro-L-Leu-Gly-L-Leu-[N3-(2,4-dinitrophenyl)-L-2,3-diamino-propionyl]-L-Ala-L-Arg-NH2
-
pH and temperature not specified in the publication
0.00449
(europium(III) complex of a modified terpyridine)-K1K2K2GFSAK2K-black hole quencher 2
-
pH and temperature not specified in the publication
-
0.65 - 0.72
Benzyloxycarbonyl-(4-nitro)Phe-Leu-Ala
-
depending on mode of preparation of the form of enzyme
0.98 - 1.39
Benzyloxycarbonyl-Gly-Phe-Leu-Ala
-
depending on mode of preparation of the form of enzyme
0.55 - 1.52
Benzyloxycarbonyl-Phe-Leu-Ala
-
depending on mode of preparation of the form of enzyme
0.77
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.38
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116Q, pH 7.5, 25°C
0.39
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant wild-type enzyme, pH 7.5, 25°C
0.41
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116A, pH 7.5, 25°C
0.48
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116D, pH 7.5, 25°C
0.49
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116T, pH 7.5, 25°C
5.8
L-phenylalanine methyl ester
-
mutant enzyme D150E, at pH 7.5, at 25°C
7
L-phenylalanine methyl ester
-
mutant enzyme D150W, at pH 7.5, at 25°C
7.3
L-phenylalanine methyl ester
-
mutant enzyme D150A, at pH 7.5, at 25°C
8.7
L-phenylalanine methyl ester
-
wild type enzyme, at pH 7.5, at 25°C
8.8
L-phenylalanine methyl ester
-
mutant enzyme N227H, at pH 7.5, at 25°C
11.3
L-phenylalanine methyl ester
-
mutant enzyme F114A, at pH 7.5, at 25°C
12.9
L-phenylalanine methyl ester
-
mutant enzyme I168A, at pH 7.5, at 25°C
0.42
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant wild-type enzyme, pH 7.5, 25°C
0.44
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117E, pH 7.5, 25°C
0.5
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117R, pH 7.5, 25°C
0.57
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117K, pH 7.5, 25°C
0.06
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.0, 25°C, recombinant mutant L144S
0.1
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P/L144S
0.16
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S/D150E/S53D, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.17
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S/D150E, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.18
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P
0.18
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme D150E/I168A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.19
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.24
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225V
0.28
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q128A
0.29
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225A
0.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutants Q128K and Q225R
0.33
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.0, 25°C, recombinant wild-type enzyme
0.33
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q128E
0.34
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225D
0.39
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225K
0.4
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant enzyme
0.4
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant wild-type enzyme
0.45
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant wild-type enzyme
0.46
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme I168A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.5
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme D150E, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.52
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, native enzyme
0.52
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, native wild-type enzyme
0.54
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.55
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/L155A/G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.63
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225E
0.63
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
wild type enzyme, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.69
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/L155A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.83
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L155A/G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.94
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
1.29
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L155A, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.7
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150E, at pH 7.5, at 25°C
0.8
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme I168A, at pH 7.5, at 25°C
1
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme F114A, at pH 7.5, at 25°C
1
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
wild type enzyme, at pH 7.5, at 25°C
1.3
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme N227H, at pH 7.5, at 25°C
2.1
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150A, at pH 7.5, at 25°C
2.1
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150W, at pH 7.5, at 25°C
49
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme F114A, at pH 7.5, at 25°C
54
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme D150W, at pH 7.5, at 25°C
57
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme D150A, at pH 7.5, at 25°C
60
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme D150E, at pH 7.5, at 25°C
62
N-carbobenzoxy-L-aspartic acid
-
wild type enzyme, at pH 7.5, at 25°C
70
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme N227H, at pH 7.5, at 25°C
105
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme I168A, at pH 7.5, at 25°C
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
thermodynamics of wild-type and mutant enzymes
-
additional information
additional information
steady-state kinetics, overview
-
additional information
additional information
-
overview: Km of synthetic oligopeptides
-
additional information
additional information
-
effect of various n-pentanal concentrations of Km
-
additional information
additional information
-
Michaelis-Menten kinetics, overview
-
additional information
additional information
-
Michaelis-Menten kinetics, overview
-
additional information
additional information
-
kinetics and thermodynamics of wild-type and mutant enzymes in absence or presence of salt, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.36
(7-methoxycoumarin-4yl) acetyl-L-Pro-L-Leu-Gly-L-Leu-[N3-(2,4-dinitrophenyl)-L-2,3-diamino-propionyl]-L-Ala-L-Arg-NH2
-
pH and temperature not specified in the publication
2.6
(europium(III) complex of a modified terpyridine)-K1K2K2GFSAK2K-black hole quencher 2
-
pH and temperature not specified in the publication
-
861 - 951
Benzyloxycarbonyl-(4-nitro)Phe-Leu-Ala
-
depending on mode of preparation of the form of enzyme
407 - 968
Benzyloxycarbonyl-Gly-Phe-Leu-Ala
-
depending on mode of preparation of the form of enzyme
362 - 605
Benzyloxycarbonyl-Phe-Leu-Ala
-
depending on mode of preparation of the form of enzyme
6.9
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
5.4
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116T, pH 7.5, 25°C
6.7
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116A, pH 7.5, 25°C
7.1
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116Q, pH 7.5, 25°C
7.7
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant wild-type enzyme, pH 7.5, 25°C
12
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116D, pH 7.5, 25°C
0.3
L-phenylalanine methyl ester
-
mutant enzyme F114A, at pH 7.5, at 25°C
1.3
L-phenylalanine methyl ester
-
wild type enzyme, at pH 7.5, at 25°C
2.1
L-phenylalanine methyl ester
-
mutant enzyme D150A, at pH 7.5, at 25°C
2.3
L-phenylalanine methyl ester
-
mutant enzyme N227H, at pH 7.5, at 25°C
2.5
L-phenylalanine methyl ester
-
mutant enzyme D150E, at pH 7.5, at 25°C
3.2
L-phenylalanine methyl ester
-
mutant enzyme D150W, at pH 7.5, at 25°C
4.9
L-phenylalanine methyl ester
-
mutant enzyme I168A, at pH 7.5, at 25°C
4.8
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117R, pH 7.5, 25°C
7.9
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant wild-type enzyme, pH 7.5, 25°C
8.4
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117K, pH 7.5, 25°C
10.5
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117E, pH 7.5, 25°C
1.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225A
1.6
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225V
3.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P
3.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225D
3.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/L155A/G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
3.4
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.0, 25°C, recombinant wild-type enzyme
3.6
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q225R
3.7
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q128K
3.8
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant enzyme
3.8
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant wild-type enzyme
3.8
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutant Q128A
4.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, native enzyme
4.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, native wild-type enzyme
4.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant mutants Q128E and Q225E
4.7
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.5, 25°C, recombinant wild-type enzyme and mutant Q225K
5.4
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
5.9
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
wild type enzyme, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
6.3
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L155A/G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
6.4
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/L155A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
6.7
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.0, 25°C, recombinant mutant L144S
7
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P/L144S
7.4
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
7.7
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme I168A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
8.7
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L155A, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
9.4
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
15.5
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme D150E/I168A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
15.6
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S/D150E/S53D, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
16.5
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme D150E, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
17
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S/D150E, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
1.5
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme F114A, at pH 7.5, at 25°C
9.5
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
wild type enzyme, at pH 7.5, at 25°C
12.3
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme I168A, at pH 7.5, at 25°C
17.1
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme N227H, at pH 7.5, at 25°C
20.5
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150E, at pH 7.5, at 25°C
24.4
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150A, at pH 7.5, at 25°C
49.4
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150W, at pH 7.5, at 25°C
0.3
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme F114A, at pH 7.5, at 25°C
1.3
N-carbobenzoxy-L-aspartic acid
-
wild type enzyme, at pH 7.5, at 25°C
2.1
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme D150A, at pH 7.5, at 25°C
2.3
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme N227H, at pH 7.5, at 25°C
2.5
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme D150E, at pH 7.5, at 25°C
3.2
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme D150W, at pH 7.5, at 25°C
4.9
N-carbobenzoxy-L-aspartic acid
-
mutant enzyme I168A, at pH 7.5, at 25°C
additional information
additional information
-
effect of various n-pentanal concentrations of turnover number
-
additional information
additional information
-
overview: turnover number of synthetic oligopeptides
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
11
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116T, pH 7.5, 25°C
16
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116A, pH 7.5, 25°C
19
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116Q, pH 7.5, 25°C
20
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant wild-type enzyme, pH 7.5, 25°C
27
N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester
purified recombinant mutant N116D, pH 7.5, 25°C
26
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
purified recombinant mutant N116T, pH 7.5, 25°C
31
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
purified recombinant mutant N116A, pH 7.5, 25°C
37
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
purified recombinant mutant N116Q, pH 7.5, 25°C
37
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
purified recombinant wild-type enzyme, pH 7.5, 25°C
117
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
purified recombinant mutant N116D, pH 7.5, 25°C
9.6
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117R, pH 7.5, 25°C
14.7
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117K, pH 7.5, 25°C
18.8
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant wild-type enzyme, pH 7.5, 25°C
23.9
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
-
recombinant mutant G117E, pH 7.5, 25°C
6
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/L155A/G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
6.7
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L155A, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
7.5
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L155A/G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
7.9
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
9.2
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme S53D/L155A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
9.4
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
wild type enzyme, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
10
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
16.8
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme I168A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
33.1
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme D150E, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
49.7
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
96.2
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S/D150E/S53D, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
99
N-carbobenzoxy-L-Asp-L-Phe methyl ester
-
mutant enzyme L144S/D150E, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
1.5
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme F114A, at pH 7.5, at 25°C
9.3
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
wild type enzyme, at pH 7.5, at 25°C
11.7
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150A, at pH 7.5, at 25°C
13.7
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme N227H, at pH 7.5, at 25°C
16
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme I168A, at pH 7.5, at 25°C
23
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150W, at pH 7.5, at 25°C
27.6
N-carbobenzoxy-L-Asp-L-Phe-methyl ester
-
mutant enzyme D150E, at pH 7.5, at 25°C
0.23
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D/L155A/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.26
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L155A, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.29
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.29
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
wild type enzyme, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.31
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.32
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.33
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D/L155A, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.34
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L155A/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.62
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme I168A, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
0.79
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme D150E, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
1.82
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L144S, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
2.04
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L144S/D150E, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
2.1
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L155A, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
2.49
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L144S/D150E/S53D, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
3
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme I168A, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
3.2
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D/L155A, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
3.3
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D/L155A/G8C/N60C/S65P, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
3.5
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme G8C/N60C/S65P, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
3.8
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
wild type enzyme, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
3.9
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
4
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme D150E, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
4
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D/G8C/N60C/S65P, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
4.3
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L155A/G8C/N60C/S65P, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
6
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L144S/D150E, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
6.6
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L144S, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
9
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme S53D, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
11.7
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
-
mutant enzyme L144S/D150E/S53D, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C
15
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
-
recombinant mutant G117R, pH 7.5, 25°C
16
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
-
recombinant mutant G117K, pH 7.5, 25°C
33
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
-
recombinant mutant G117E, pH 7.5, 25°C
40
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
-
recombinant wild-type enzyme, pH 7.5, 25°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.57
2-(acetyloxy)-3-chlorobenzoic acid
in 100 mM Tris/HCl, pH 7.5, 2 mM CaCl2, 4% (v/v) DMSO, at 25°C
0.315
2-ethyl-3-hydroxyquinazolin-4(3H)-one
in 100 mM Tris/HCl, pH 7.5, 2 mM CaCl2, 4% (v/v) DMSO, at 25°C
0.128
3-hydroxy-2-isopropylquinazolin-4(3H)-one
in 100 mM Tris/HCl, pH 7.5, 2 mM CaCl2, 4% (v/v) DMSO, at 25°C
0.93
3-hydroxy-2-methylquinazolin-4(3H)-one
in 100 mM Tris/HCl, pH 7.5, 2 mM CaCl2, 4% (v/v) DMSO, at 25°C
1.7
3-methylaspirin
in 100 mM Tris/HCl, pH 7.5, 2 mM CaCl2, 4% (v/v) DMSO, at 25°C
0.000000068
carbobenzoxy-L-phenylalanine-phosphonamidate-L-leucyl-L-alanine
pH and temperature not specified in the publication
0.00005
N-(1-carboxy-3-phenyl-propyl)-L-leucyl-L-tryptophan
pH and temperature not specified in the publication
0.0012
[(2S)-2-sulfanyl-3-phenylpropanoyl]Gly-(5-Ph)Pro
pH 8.0, 37°C
0.000042
[(2S)-2-sulfanyl-3-phenylpropanoyl]Phe-Tyr
pH 8.0, 37°C
0.000048
[(2S,R)-2-sulfanylheptanoyl]Phe-Ala
pH 8.0, 37°C
0.000000064
1-beta-D-arabinofuranosyl-N4-lauroylcytosine
-
25°C, pH 7.5
0.0593
2-(4-methylphenyl)-3-(1,3-thiazol-2-yl) quinazolin-4(3H)-one
-
at pH 7.0 and 25°C
2.06
2-hydroxy-N-(4-methyl-2-nitrophenyl)-3-nitrobenzamide
-
25°C, pH 7.5
0.000243
3-(isopropylideneamino)-2,2-dimethyl-2,3-dihydroquinazolin-4(1H)-one
-
at pH 7.0 and 25°C
0.0183
3-([(1E)-[4-(dimethylamino) phenyl]methylene]amino)-2-methylquinazolin-4(3H)-one
-
at pH 7.0 and 25°C
0.042
3-([(1E)-[4-(dimethylamino) phenyl]methylene]amino)-2-phenylquinazolin-4(3H)-one
-
at pH 7.0 and 25°C
10100
3-amino-2-(4-chlorophenyl)quinazolin-4(3H)-one
-
at pH 7.0 and 25°C
3.12
3-amino-2-(4-nitrophenyl)quinazolin-4(3H)-one
-
at pH 7.0 and 25°C
0.0379
3-amino-2-(trifluoromethyl) quinazolin-4(3H)-one
-
at pH 7.0 and 25°C
0.0549
3-amino-2-methylquinazolin-4(3H)-one
-
at pH 7.0 and 25°C
0.0000115
3-phenyl-2-(trifluoromethyl) quinazolin-4(3H)-one
-
at pH 7.0 and 25°C
4
3-[[(1E)-(3-chlorophenyl)methylene]amino]-2-phenylquinazolin-4(3H)-one
-
at pH 7.0 and 25°C
4.81
5-bromo-N-(4-bromophenyl)-2-hydroxy-3-nitro-benzamide
-
25°C, pH 7.5
1.6
beta-phenylpropionyl-L-phenylalanine
-
pH 6.8, 25°C, enzyme contains Zn2+
106
ethanimidic acid N-[4-oxo-2-phenyl-3(4H)-quinazolinyl]-ethyl ester
-
at pH 7.0 and 25°C
294
N-(2,3-dimethylphenyl)-2-hydroxy-3-nitro-benzamide
-
25°C, pH 7.5
161
N-(2,4-dimethylphenyl)-2-hydroxy-3-nitro-benzamide
-
25°C, pH 7.5
1.45
N-(2,5-dimethylphenyl)-2-hydroxy-3-nitrobenzamide
-
25°C, pH 7.5
697
N-(2-chloro-4-nitrophenyl)-2-hydroxy-3-nitro-benzamide
-
25°C, pH 7.5
0.047
N-(2-chloro-6-methylphenyl)-2-hydroxy-3-nitrobenzamide
-
25°C, pH 7.5
2.95
N-(2-ethylphenyl)-2-hydroxy-3-nitrobenzamide
-
25°C, pH 7.5
8.14
N-(5-chloro-2-methoxyphenyl)-2-hydroxy-3-nitro-benzamide
-
25°C, pH 7.5
0.019
Z-L-tryptophan
-
pH 5.0, 25°C, enzyme contains Zn2+, presence of 1 M NaSCN
0.021
Z-L-tryptophan
-
pH 5.0, 25°C, enzyme contains Co2+, presence of 1 M NaSCN
additional information
additional information
-
detailed inhibition kinetics with Co2+ and Zn2+, overview
-
additional information
additional information
-
inhibition kinetics with Co2+ and Ca2+, overview
-
additional information
additional information
-
organic solvent inhibition kinetics, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.07685
2-(4-methylphenyl)-3-(1,3-thiazol-2-yl) quinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
0.067
2-[(biphenyl-4-ylsulfonyl)[2-(hydroxyamino)-2-oxoethyl]amino]-N-[2-(4-sulfamoylphenyl)ethyl]acetamide (non-preferred name)
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.012
2-[benzyl[2-(hydroxyamino)-2-oxoethyl]amino]-N-[2-(4-sulfamoylphenyl)ethyl]acetamide (non-preferred name)
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.008
2-[benzyl[2-(hydroxyamino)-2-oxoethyl]amino]-N-[3-(4-phenylpiperazin-1-yl)propyl]acetamide (non-preferred name)
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.164
2-[[2-(hydroxyamino)-2-oxoethyl][(4-methoxyphenyl)sulfonyl]amino]-N-[2-(4-sulfamoylphenyl)ethyl]acetamide (non-preferred name)
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.16
2-[[2-(hydroxyamino)-2-oxoethyl][(4-phenoxyphenyl)sulfonyl]amino]-N-[2-(4-sulfamoylphenyl)ethyl]acetamide (non-preferred name)
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.0002477
3-(isopropylideneamino)-2,2-dimethyl-2,3-dihydroquinazolin-4(1H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
0.01832
3-([(1E)-[4-(dimethylamino) phenyl]methylene]amino)-2-methylquinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
0.04203
3-([(1E)-[4-(dimethylamino) phenyl]methylene]amino)-2-phenylquinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
12.74
3-amino-2-(4-chlorophenyl)quinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
3.118
3-amino-2-(4-nitrophenyl)quinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
0.03786
3-amino-2-(trifluoromethyl) quinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
0.05489
3-amino-2-methylquinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
0.0000115
3-phenyl-2-(trifluoromethyl) quinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
4.002
3-[[(1E)-(3-chlorophenyl)methylene]amino]-2-phenylquinazolin-4(3H)-one
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
122.6
ethanimidic acid N-[4-oxo-2-phenyl-3(4H)-quinazolinyl]-ethyl ester
Bacillus thermoproteolyticus
-
at pH 7.0 and 25°C
1
N-[(2R)-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl]-N-[(4-phenoxyphenyl)sulfonyl]glycine
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
1
[(biphenyl-4-ylmethyl)[2-(hydroxyamino)-2-oxoethyl]amino]acetic acid
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.44
[(biphenyl-4-ylsulfonyl)[2-(hydroxyamino)-2-oxoethyl]amino]acetic acid
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
1
[1-[2-(hydroxyamino)-2-oxoethyl]-2-[3-(4-phenylpiperazin-1-yl)propyl]hydrazinyl]acetic acid
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.637
[[(4-methoxyphenyl)sulfonyl](2-oxo-2-[[2-(4-sulfamoylphenyl)ethyl]amino]ethyl)amino]acetic acid
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.755
[[2-(hydroxyamino)-2-oxoethyl](4-nitrobenzyl)amino]acetic acid
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.524
[[2-(hydroxyamino)-2-oxoethyl](4-phenoxybenzyl)amino]acetic acid
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.27
[[2-(hydroxyamino)-2-oxoethyl][(4-methoxyphenyl)sulfonyl]amino]acetic acid
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
0.114
[[2-(hydroxyamino)-2-oxoethyl][(4-phenoxyphenyl)sulfonyl]amino]acetic acid
Bacillus thermoproteolyticus
-
pH 7.5, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
12000
purified recombinant wild-type enzyme, pH 7.5, 25°C, substrate N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
5300
purified recombinant mutant N116A, pH 7.5, 25°C, substrate N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
5400
purified recombinant mutant N116T, pH 7.5, 25°C, substrate N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
6800
purified recombinant mutant N116Q, pH 7.5, 25°C, substrate N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
7900
purified recombinant mutant N116D, pH 7.5, 25°C, substrate N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide
0.42
0.89
10.2
-
purified mutant enzyme S53D/L155A, using casein as substrate, at pH 7.5 and 25°C
10.5
-
purified mutant enzyme D150E, using casein as substrate, at pH 7.5 and 25°C
10.8
11.3
-
purified mutant enzyme S53D, using casein as substrate, at pH 7.5 and 25°C
11.9
-
purified mutant enzyme S53D/G8C/N60C/S65P, using casein as substrate, at pH 7.5 and 25°C
11000
-
recombinant wild-type enzyme, substrate casein, pH 7.5, 25°C
12000
-
recombinant mutant G117E, substrate casein, pH 7.5, 25°C
3.5
-
purified mutant enzyme L144S/D150E, using casein as substrate, at pH 7.5 and 25°C
3.6
-
purified mutant enzyme L144S, using casein as substrate, at pH 7.5 and 25°C
6.7
-
purified mutant enzyme L155A, using casein as substrate, at pH 7.5 and 25°C
7.5
-
purified mutant enzyme D150E/I168A, using casein as substrate, at pH 7.5 and 25°C
7.9
-
purified mutant enzyme S53D/L155A/G8C/N60C/S65P, using casein as substrate, at pH 7.5 and 25°C
8.1
-
purified mutant enzyme L155A/G8C/N60C/S65P, using casein as substrate, at pH 7.5 and 25°C
9.6
-
purified mutant enzyme I168A, using casein as substrate, at pH 7.5 and 25°C
additional information
10.8
-
purified mutant enzyme G8C/N60C/S65P, using casein as substrate, at pH 7.5 and 25°C
10.8
-
purified wild type enzyme, using casein as substrate, at pH 7.5 and 25°C
additional information
one proteolytic unit is defined as the amount which liberates a quantity of acid-soluble peptides that corresponds to an increase in A275 of 0.0074 (A275 of 1 mg of tyrosine)/min
additional information
-
catalytic efficiency of wild-type and mutant enzymes in absence or presence of 4 M NaCl, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.5 - 8.5
for hydrolysis of neutral substrate N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide, wild-type and mutants N112D, N112E exhibit bell-shaped pH-dependence. For hydrolysis of negatively charged substrate N-carbobenzoxy-L-Asp-L-Phe methyl ester, wild-type shows bell-shaped pH-dependence, for mutants N112D and N112E, pH-dependence of the ratio kcat/Km decreases with increase in pH from 5.5 to 8.5
additional information
additional information
-
bell-shaped pH-dependence with importance of surfacecharges of thermolysin, the bell-shaped pH dependence profile of the FAGLA-hydrolyzing activity of thermolysin is shifted from pH 5.4 to pH 6.7 by the addition of 4 M NaCl
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25 - 70
-
hydrolysis of bovine alpha-lactalbumin at 25°C and 70°C under nonreducing conditions. At 25°C, substrate undergoes limited hydrolysis leading to peptides no longer degraded. At 70°C, protein is first quickly cleaved, then unfolded, leading to the release of intermediate peptides that may be further degraded
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
-
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
pro-enzyme form, folding and autocleavage takes place in the periplasmic space, overview
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
thermolysin and pseudolysin belong to subclan MA(E) of peptidases, also known as the Glu-zincins, of the matrix metalloproteinases family of zinc-containing endoproteinases with broad substrate specificity and extracellular matrix components are among the substrates
physiological function
-
thermolysin family proteases are related to various diseases such as bacterial infections, cholera, gastritis and peptic ulcers, and gastric carcinoma
additional information
additional information
proposed mechanism of action for the thermolysin-catalyzed synthesis of aspartame precursor carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, overview. Residues E143, Y157, and H231 represent the catalytic triad, enzyme active site structure analysis
additional information
structure analsis using enzyme structure PDB ID 8TLN. Catalytically important residues Glu143 and His231. Important role of Asn116 in the activity and stability of thermolysin presumably by stabilizing the beta-hairpin structure. In the N-terminal domain of thermolysin, two antiparallel beta-strands, Asn112-Ala113-Phe114-Trp115 and Ser118-Gln119-Met120-Val121-Tyr122 are connected by an Asn116-Gly117 turn to form a beta-hairpin structure
additional information
-
development and evaluation of a method providing information at the surface of the outer envelope membrane, based on specific tagging with biotin or proteolysis using thermolysin, a non-membrane permeable protease. Envelope, thylakoid, and stroma proteins are separated by two-dimensional electrophoresis and analyzed by immunostaining and mass spectrometry, overview
additional information
-
the glutamic acid of the HEXXH motif is catalytically important
additional information
-
the peptide products from hydrolysis of elastin and collagen by the enzyme have an inhibitory effect on angiotensin I-converting enzyme and antihypertensive activity, overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
34600
26000
-
x * 22 852, propeptide of thermolysin, sequence calculation, x * 26000, mature enzyme, SDS-PAGE
34600
34800
-
Bacillus thermoproteolyticus, amino acid composition, amino acid sequence
37500
-
Bacillus thermoproteolyticus, sedimentation equilibrium method
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
-
1 * 37000-38000, Bacillus thermoproteolyticus, SDS-PAGE, gel filtration in 6 M guanidine hydrochloride
additional information
?
-
x * 22 852, propeptide of thermolysin, sequence calculation, x * 26000, mature enzyme, SDS-PAGE
additional information
-
in presence of Ca2+, enzyme exhibits four autodegradation sites, one of them being Gly154-Leu155
additional information
-
structure determination and molecular dynamics simulations, overview
additional information
-
the enzyme consists of a beta-rich N-terminal domain and an alpha-helix C-terminal domain connected by a central alpha-helix, which is located at the bottom of the active site cleft
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
proteolytic modification
-
the pre-pro-enzyme contains a signal peptide and a prosequence, the prosequence acts as an intramolecular chaperone for autocatalytic cleavage of the linking peptide bond
proteolytic modification
-
thermolysin is synthesized as inactive pre-proenzyme and receives autocatalytic cleavage of the peptide bond linking the pro- and mature sequences, overview
proteolytic modification
-
thermolysin performs Co2+-stimulable autolysis
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
analysis of the crystal structure of enzyme complexed with substrates L-phenylalanine methyl ester and carbobenzoxy-L-aspartic, or with inhibitor carbobenzoxy-L-phenylalanine-phosphonamidate-L-leucyl-L-alanine, X-ray diffraction structure determination and analysis, modeling
native thermolysin in complex with 3-methylaspirin, 3-methylaspirin ethyl ester, and 3-methylaspirin cyclopropyl ester, sitting drop vapor diffusion method, using water as reservoir solution, at 18°C
thiocyanate as crystallizing agent, space group P6(1)22, unit-cell parameters a and b : 93.17 A
TLN-inhibitor complex, 3 alpha-mercaptoacyldipeptides in the thermolysin active site crystallized, hexagonal crystals, space group P6(1)22, [(2S)-2-sulfanyl-3-phenylpropanoyl]Gly-(5-Ph)Pro, parameters a : b : 93.50 A, c : 131.30 A, [(2S)-2-sulfanyl-3-phenylpropanoyl]Phe-Tyr, a : b : 93.97 A, c : 131.79 A, [(2S,R)-2-sulfanylheptanoyl]Phe-Ala, a : b : 93.31 A, c : 131.83 A
crystallization of tetragonal thermolysin, at room temperature, structure determination and anaylsis, fluctuations and solvent-accessible surface areas, biological nanopores and water densities, and radial distributions of water and ions, overview
-
data in presence of 4 M NaCl, introduced into crystals originally grown without NaCl
-
multiple-solvent crystal structure determination cell dimensions dependent on isopropanol concentration
-
resiudes H142, H146, and E166 coordinate the catalytic zinc, while E143 and H231 are required for the catalytic activity
-
three times crystallized and lyophilized preparation, Lot T5CB491
-
three-times crystallized preparation, Lot T5CB491 from Daiwa Kasei, Osaka
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
N112A
no enzymic activity in supernatant of cells expressing mutant
N112D
supernatants of cells expressing mutant show 18% of wild-type activity
N112E
supernatants of cells expressing mutant show 5% of wild-type activity
N112H
no enzymic activity in supernatant of cells expressing mutant
N112K
no enzymic activity in supernatant of cells expressing mutant
N112R
no enzymic activity in supernatant of cells expressing mutant
N116A
site-directed mutagenesis, the mutant shows slightly decreased activity compared to the wild-type enzyme
N116D
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
N116Q
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
N116T
site-directed mutagenesis, the mutant shows slightly decreased activity compared to the wild-type enzyme
S198D
site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme
S218D
site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme
S254D
site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme
S25D
site directed mutagenesis, the catalytic activity is of the mutant enzyme is similar to the wild-type in absence of NaCl, but increased in presence of 4 M NaCl
S53D
site directed mutagenesis, the catalytic activity is of the mutant enzyme is similar to the wild-type in absence of NaCl, but increased in presence of 4 M NaCl, increased thermostability in presence of 10 mM CaCl2
S65D
site directed mutagenesis, the catalytic activity is of the mutant enzyme is similar to the wild-type in absence of NaCl, but increased in presence of 4 M NaCl, increased thermostability in presence of 10 mM CaCl2
A4T/G8C/T56A/G58A/N60C/T63F/S65P/A69P
-
the mutant shows altered thermodynamics
A4T/T56A/G58A/T63F/S65P/A69P
-
the mutant shows altered thermodynamics
D150A
D150E
D150H
-
105% residual activity with casein, 37% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed
D150K
-
51% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
D150R
-
44% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
D150W
D170A
DELTA127
-
absence of CaCl2, 18% of wild-type activity, presence of 5 mM CaCl2, 71% of wild-type activity. Decrease in amount of enzyme secreted compared to wild-type
E143A
-
site-directed mutagenesis, E143A might exist as a complex with the propetide in the supernatant, inactive mutant, the autocatalytic activity is affected
F114A
F114H
-
18% residual activity with casein, 20% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
G117D
-
site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type
G117E
-
site-directed mutagenesis, the mutant enzyme shows increased activity compared to the wild-type enzyme, the kcat/Km value is 80% of wild-type level with N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide, but 130% with N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
G117K
-
site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type, the kcat/Km value is 40% of wild-type level with N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide, but 80% with N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
G117R
-
site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type, the kcat/Km value is 40% of wild-type level with N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide and N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester
G8C/N60C/S65P
G8C/N60C/S65P/L144S
H231A
-
the mutant shows 500fold decreased catalytic efficiency compared to the wild-type enzyme
I168A
I168H
-
13% residual activity with casein, 35% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed
L144S
L144S/D150E
-
the mutation yields the most significant increase in the hydrolytic activities for N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide and N-carbobenzoxy-L-Asp-L-Phe methyl ester and shows about 30% casein-hydrolytic activity compared to the wild type enzyme
L144S/D150E/S53D
-
the triple mutant shows improved activity and stability with about 30% casein-hydrolytic activity compared to the wild type enzyme
L144S/D150W/N227H
-
the mutant shows 10fold decreased catalytic efficiency compared to the wild-type enzyme
L144S/I168A
-
the mutation abolishes the hydrolytic activities for N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide and N-carbobenzoxy-L-Asp-L-Phe methyl ester
L155A
L155A/G8C/N60C/S65P
-
the mutant shows about 80% casein-hydrolytic activity compared to the wild type enzyme
L155F
-
thermostability at 80°C increases with amino acid substitutions at L155 in the order of wild-type, Gly, Ser, Phe, Ala. Autodegradation site shifts from G154-L155 to F155-I156 and the bond I164-D165 is newly recognized as an autodegradation site
L155G
-
thermostability at 80°C increases with amino acid substitutions at L155 in the order of wild-type, Gly, Ser, Phe, Ala. Autodegradation site shifts from G154-L155 to G155-I156 and the bond I164-D165 is newly recognized as an autodegradation site
L155S
M205P
-
absence of CaCl2, 0.52% of wild-type activity, presence of 5 mM CaCl2, 48% of wild-type activity. Decrease in amount of enzyme secreted compared to wild-type
N112A
-
site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected
N112D
N112E
-
site-directed mutagenesis, the autocatalytic activity is affected
N112H
-
site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected
N112K
-
site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected
N112R
-
site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected
N116D/Q119R/D150Q/Q225R
-
the mutant shows 4fold decreased catalytic efficiency compared to the wild-type enzyme
N227A
-
72% residual activity with casein , 28% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed
N227D
-
11% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
N227E
-
36% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
N227H
N227K
-
29% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
N227R
-
55% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
P208H
-
absence of CaCl2, 0.61% of wild-type activity, presence of 5 mM CaCl2, 61% of wild-type activity. Decrease in amount of enzyme secreted compared to wild-type
Q128A
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
Q128E
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme
Q128K
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Q225A
-
site-directed mutagenesis, the mutant shows altered pKa value and stimulation of activity by NaCl and reduced activity with the negatively charged substrate N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester substrate compared to the wild-type enzyme
Q225D
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
Q225E
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme
Q225K
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
Q225R
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme
Q225V
-
site-directed mutagenesis, the mutant shows altered pKa value and stimulation of activity by NaCl and reduced activity with the negatively charged substrate N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester substrate compared to the wild-type enzyme
R203A
-
the mutant shows 5fold decreased catalytic efficiency compared to the wild-type enzyme
R203M
S103A
-
the mutant shows 3fold decreased catalytic efficiency compared to the wild-type enzyme
S169A
-
112% residual activity with casein, 64% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
S234A
-
88% residual activity with casein, 17% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed
S234D
-
5% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
S234E
-
4% residual activity with casein, 7% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
S234H
-
32% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
S53D/G8C/N60C/S65P
-
the mutant shows about 110% casein-hydrolytic activity compared to the wild type enzyme
S53D/L155A
-
the mutation yields the greatest increase in the thermal stability and shows about 90% casein-hydrolytic activity compared to the wild type enzyme
S53D/L155A/G8C/N60C/S65P
-
the mutant shows about 70% casein-hydrolytic activity compared to the wild type enzyme
V230A
-
17% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
V230K
-
3% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
V230R
-
6% residual activity with casein, 12% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157A
-
11% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157D
-
1% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157E
-
13% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157H
-
24% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157K
-
7% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
additional information
D150A
-
131% residual activity with casein, 81% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
D150A
-
mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type
D150E
-
128% residual activity with casein, 228% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
D150E
-
mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type
D150W
-
81% residual activity with casein, 60% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed
D150W
-
mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type
F114A
-
28% residual activity with casein, 8% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
G8C/N60C/S65P
-
site-directed mutagenesis, the mutant shows a similar catalytic efficiency compared tot he wild-type enzyme
G8C/N60C/S65P
-
the mutation increases the stability of thermolysin
G8C/N60C/S65P
-
the triple mutation increases the stability of thermolysin as high as the individual mutations do
G8C/N60C/S65P/L144S
-
site-directed mutagenesis, the mutant shows about 6fold increased catalytic effiency compared to the wild-type enzyme
G8C/N60C/S65P/L144S
-
the mutant is more active and stable than wild type thermolysin
I168A
-
69% residual activity with casein, 125% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed
I168A
-
mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type
I168A
-
the mutation increases the activity of thermolysin and shows about 90% casein-hydrolytic activity compared to the wild type enzyme
L144S
-
site-directed mutagenesis, the mutant shows about 10fold increased catalytic effiency compared to the wild-type enzyme
L144S
-
the mutation increases the activity of thermolysin and shows about 30% casein-hydrolytic activity compared to the wild type enzyme
L155A
-
absence of CaCl2, 87% of wild-type activity, presence of 5 mM CaCl2, 83% of wild-type activity. Amount of enzyme secreted is about the same level as wild-type
L155A
-
thermostability at 80°C increases with amino acid substitutions at L155 in the order of wild-type, Gly, Ser, Phe, Ala. Autodegradation site shifts from G154-L155 to A155-I156 and the bond I164-D165 is newly recognized as an autodegradation site
L155A
-
the mutation increases the stability of thermolysin and shows about 60% casein-hydrolytic activity compared to the wild type enzyme
L155S
-
thermostability at 80°C increases with amino acid substitutions at L155 in the order of wild-type, Gly, Ser, Phe, Ala. Autodegradation site shifts from G154-L155 to S155-I156 and the bond I164-D165 is newly recognized as an autodegradation site
L155S
-
the mutant shows increased stability at 80°C compared to the wild-type enzyme
N112D
-
site-directed mutagenesis, the autocatalytic activity is affected
N227H
-
19% residual activity with casein, 19% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed
N227H
-
mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type
R203M
-
the mutant shows 2300fold decreased catalytic efficiency compared to the wild-type enzyme
additional information
-
a mutant thermolysin is affected by its autocatalytic digestion activity
additional information
-
generation of an engineered enzyme with a higher activity in the synthesis of N-carbobenzyloxy L-Asp-L-Phe methyl ester
additional information
-
evaluation of an efficient method for the immobilization of thermolysin using sodium chloride salting-in and consecutive microwave irradiation, overview. 4.6% of the relative activity for the immobilized thermolysin is detected when the immobilization mixture contains no salts, including ZnCl2, CaCl2 or NaCl
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7
-
after precipitation with trichloroacetic acid the purified enzyme irreversibly loses activity and solubility at neutral pH
31136
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
80
first-order rate constant of the thermal inactivation at 80°C in the presence of 1-100mM CaCl2
85
30 min, 51% remaining activity of recombinaqnt wild-type enzyme, 35-78% remaining activity of mutant enzymes
80
80
-
1 h, heated enzyme has the same maximum velocity but lower affinity for substrate than the native
80
-
thermoinactivation of thermolysin at 80°C in 50% of solvents, half-lives of 3-20 min, with the increase in solvent hydrophobicity, thermal stability of the enzyme decreases, overview
80
-
the rate constant (kobs) for thermal inactivation at 80°C is 84000 s-1 for the wild type enzyme
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
important role of Asn116 in the activity and stability of thermolysin presumably by stabilizing the beta-hairpin structure. In the N-terminal domain of thermolysin, two antiparallel beta-strands, Asn112-Ala113-Phe114-Trp115 and Ser118-Gln119-Met120-Val121-Tyr122 are connected by an Asn116-Gly117 turn to form a beta-hairpin structure
After precipitation with trichloroacetic acid the purified enzyme irreversibly loses activity and solubility at neutral pH
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2-methyl-2-butanol
-
no reduction in activity of immobilized thermolysin after 2 h in 5% tert-amyl alcohol, whereas the free enzyme loses 32.5% of its catalytic activity
dimethylformamide
-
half-life is 6 min at 80°C, trehalose stabilizes the enzyme in DMF
n-propanol
-
half-life is 3 min at 80°C, glycerol stabilizes the enzyme
additional information
-
solvents cause mixed inhibition of thermolysin, kinetic and structural studies, overview
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
commercial enzyme powder further purified to remove salts
recombinant mature wild-type and mutant enzymes 7.2-11fold from Escherichia coli by hydrophobic interaction and affinity chromatography
recombinant wild-type and mutant enzymes from Escherichia coli K12 strain JM109 to homogeneity by Gly-D-Phe heat treatment at 60°C for 20 min, affinity chromatography and gel filtration
by Gly-D-Phe affinity chromatography, coupling to the resin by epichlorohydrin, 1,4-butandiol diglycidyl ether, or 1,6-hexanediol diglycidyl ether, method optimization and evaluation, overview
-
evaluation of Gly-D-Phe, Gly-L-Leu, and D-Phe as affinity ligands for thermolysin, each of the ligands is immobilized to a resin. The optimum pH for adsorption of thermolysin is pH 5.0 to pH 6.0 for each of the ligands, affinity chromatography method development, adsorption isotherms, overview
-
expression as single polypeptide pre-proenzyme in Escherichia coli, secretion into medium as mature enzyme
-
mobile phase effects in the high-performance affinity purification
-
recombinant wild-type and mutant enzymes from Escherichia coli K12 strain JM109 by hydrophobic interaction and Gly-D-Phe affinity chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of wild-type and mutant enzymes in Escherichia coli K12 strain JM109
recombinant overexpression of wild-type and mutant enzymes in Escherichia coli
cloned in Bacillus subtilis DB117, plasmids subcloned in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli K12 strain JM109 and secretion to the cell culture medium
-
expression of wild-type and mutant enzymes in Escherichia coli strain T5KC991
-
gene npr, DNA and amino acid sequence determination, expression of wild-type and mutant enzymes in Escherichia coli K12 strain JM109
-
gene npr, expression of wild-type and mutant enzymes in Escherichia coli K12 strain JM109
-
genes npr and nprT, DNA and amino acid sequence determination and analysis, expression in Escherichia coli and Bacillus subtilis
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
thermolysin degrades cellular prion protein while preserving both proteinase K-sensitive and proteinase K-resistant isoforms of disease-related prion protein in both rodent and human prion strains. In variant Creutzfeldt-Jakob disease, up to 90% of total prion protein present in the brain resists degradation with thermolysin, whereas only about 15% of this material resists digestion by proteinase K
medicine
thermolysin degrades cellular prion protein while preserving both proteinase K-sensitive and proteinase K-resistant isoforms of disease-related prion protein in both rodent and human prion strains. In variant Creutzfeldt-Jakob disease, up to 90% of total prion protein present in the brain resists degradation with thermolysin, whereas only about 15% of this material resists digestion by proteinase K
synthesis
analysis
diagnostics
-
thermolysin is used in the diagnosis of prion diseases ovine scrapie and bovine spongiform encephalopathy, with similar sensitivity compared to proteinase K digestion, use of a protease to distinguish PrPC from PrPSc, overview
food industry
-
the enzyme can be used for production of caseicin A, an antimicrobial active peptide, from alpha-casein, for potential improvement of the safety of infant milk formula using milk-derived bioactive peptides
nutrition
-
the enzyme is used for synthesis of N-carbobenzyloxy L-Asp-L-Phe methyl ester, a precursor of the artificial sweetener aspartam
synthesis
additional information
synthesis
introduction of ionizing residues into the active site of enzyme as a means of modifying its pH-activity profile
synthesis
using the enzyme to catalyze the condensation of the chiral aspartame-precursor, carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, from the protected amino acid substrates carbobenzoxy-L-aspartic acid and L-phenylalanine methyl ester in large scale production. Analysis of the protease mediated peptide synthesisof a precursor of the artificial sweetener aspartame, a multiton peptide synthesis catalyzed by the enzyme thermolysin. X-ray structures of thermolysin in complex with aspartame substrates separately, and after protease mediated peptide synthesis in a crystal, rationalize the reaction's substrate preferences and reveal an unexpected form of substrate inhibition that explains its sluggishness. Structure guided optimization of this and other PMPS reactions could expand the economic viability of commercial peptides beyond current high-potency, low-volume therapeutics, with substantial green chemistry advantages
synthesis
sequential hydrolysis trypsin-thermolysin of commercial hydrolysate (prepared from solubilized milk proteins) has been performed to produce bioactive peptides, mainly casein phosphopeptides. Thermolysin was the last enzyme to be used, generating the final product, with a low content of aromatic amino acids and shorter peptides
analysis
-
selective biotin tagging and thermolysin proteolysis of chloroplast outer envelope proteins reveals information on protein topology and association into complexes. Development and evaluation of a method providing information at the surface of the outer envelope membrane, based on specific tagging with biotin or proteolysis using thermolysin, a non-membrane permeable protease. Envelope, thylakoid, and stroma proteins are separated by two-dimensional electrophoresis and analyzed by immunostaining and mass spectrometry, overview
analysis
-
thermolysin offers a tool for complete solubilization of cartilage prior to comprehensive glycosaminoglycans(GAG)omic analysis, and is likely applicable to other collagen-rich tissues such as ligaments, skin, and blood vessels
synthesis
-
hydrolysis and condensation reactions of peptides catalyzed by enzyme can be reversibly controlled by on/off ultrasound irradiation depending on its frequency region
synthesis
-
immobilized enzyme catalyzes the formation of beta-cyclodextrin esters using vinyl esters of butyrate, decanoate and laurate, as acyl donors in dimethylsulfoxide. Esterification occurs exclusively at the glucose C-2 position. Enzyme also catalyzes the synthesis of alpha-, beta-, gamma- and maltosyl-beta-cyclodextrin esters with vinyl laurate as the acyl donor in dimethylsulfoxide and dimethylformamide
synthesis
-
immobilized enzyme catalyzes the transesterification of vinyl laurate to several sucrose-containing tri- and tetrasaccharides. Preferred position of acylation is the 2-OH group of the alpha-D-glucopyranose moiety linked 1 to 2 to the beta-D-fructofuranose unit
synthesis
-
the enzyme is used for synthesis of N-carbobenzyloxy L-Asp-L-Phe methyl ester, a precursor of the artificial sweetener aspartam
synthesis
-
thermolysin is industrially used for the synthesis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, a precursor of an artificial sweetener aspartame, from N-carbobenzoxy-L-aspartic acid and L-phenylalanine methyl ester
synthesis
-
the enzyme can be used for production of caseicin A, an antimicrobial active peptide, from alpha-casein, for potential improvement of the safety of infant milk formula using milk-derived bioactive peptides
additional information
-
thermolysin crystal can be used as the stationary phase in liquid chromatography to separate D/L-phenylglycine, thermolysin crystal is useful for chiral separation
additional information
-
thermolysin treatment improves the yield of human intestinal epithelial cells. The thermolysin and endothelin-3 method can be used to isolate and generate viable human intestinal epithelial cells from the human small intestine
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
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Comparative study of various neutral proteinases from microorganisms: specificity with oligopeptides
Arch. Biochem. Biophys.
146
291-296
1971
Aspergillus oryzae, Bacillus thermoproteolyticus
Ohta, Y.; Ogura, Y.; Wada, A.
Thermostable protease from thermophilic bacteria. I. Thermostability, physiocochemical properties, and amino acid composition
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241
5919-5925
1966
Bacillus thermoproteolyticus
Nurul Alam, M.; Tadasa, K.; Kayahara, H.
Kinetic behavior of activation of thermolysin by normal alcohols
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18
45-50
1996
Bacillus thermoproteolyticus
-
Inagaki, T.; Tadasa, K.; Kayahara, H.
Modification effects of organic solvents on microenvironment of the enzyme in thermolysin-catalyzed peptide synthesis of N-(benzyloxycarbonyl)-L-phenylalanyl-L-phenylalanine methyl ester
Biosci. Biotechnol. Biochem.
58
1439-1442
1994
Bacillus thermoproteolyticus
-
Marcotte, P.A.; Henkin, J.
Characterization of the activation of pro-urokinase by thermolysin
Biochim. Biophys. Acta
1161
105-112
1993
Bacillus thermoproteolyticus
Zamai, M.; Fassina, G.
Mobile phase effects in the high-performance affinity purification of thermolysin
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549
195-205
1991
Bacillus thermoproteolyticus
-
Yang, J.J.; van Wart, H.E.
Kinetics of hydrolysis of dansyl peptide substrates by thermolysin: analysis of fluorescence changes and determination of steady-state kinetic parameters
Biochemistry
33
6508-6515
1994
Bacillus thermoproteolyticus
Gettins, P.
Thermolysin-inhibitor complexes examined by 31P and 113Cd NMR spectroscopy
J. Biol. Chem.
263
10208-10211
1988
Bacillus thermoproteolyticus
Grobelny, D.; Poncz, L.; Galardy, R.E.
Inhibition of human skin fibroblast collagenase, thermolysin, and Pseudomonas aeruginosa elastase by peptide hydroxamic acids
Biochemistry
31
7152-7154
1992
Bacillus thermoproteolyticus
Christianson, D.W.; Lipscomb, W.N.
Comparison of carboxypeptidase A and thermolysin: inhibition by phosphonamidates
J. Am. Chem. Soc.
110
5560-5565
1988
Bacillus thermoproteolyticus
-
Kitagishi, K.; Hiromi, K.
Binding between thermolysin and its specific inhibitor, N-phosphoryl-L-leucyl-L-tryptophan (PLT)
J. Biochem.
99
191-197
1986
Bacillus thermoproteolyticus
Kitagishi, K.; Hiromi, K.
Binding between thermolysin and its specific inhibitor, phosphoramidon
J. Biochem.
95
529-534
1984
Bacillus thermoproteolyticus
Maycock, A.L.; DeSousa, D.M.; Payne, L.G.; ten Broeke, J.; Wu, M.T.; Patchett, A.A.
Inhibition of thermolysin by N-carboxymethyl dipeptides
Biochem. Biophys. Res. Commun.
102
963-969
1981
Bacillus thermoproteolyticus
Gray, R.D.; Pierce, W.M.; Harrod, J.W.; Rademacher, J.M.
Inhibition of thermolysin by bifunctional N-carboxyalkyl dipeptides
Arch. Biochem. Biophys.
256
692-698
1987
Bacillus thermoproteolyticus
Durrant, I.; Beynon, R.J.; Rodgers, P.B.
The effect of inhibitors on thermolysin-catalyzed peptide bond synthesis
Biochem. Soc. Trans.
14
143
1986
Bacillus thermoproteolyticus
-
Nakanishi, K.; Kimura, Y.; Matsuno, R.
Kinetics and equilibrium of enzymatic synthesis of peptides in aqueous/organic biphasic systems. Thermolysin-catalyzed synthesis of N-(benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester [published erratum appears in Eur J Biochem 1987 Sep 15;167(3):601]
Eur. J. Biochem.
161
541-549
1986
Bacillus thermoproteolyticus
Feder, J.; Brougham, L.R.; Wildi, B.S.
Inhibition of thermolysin by dipeptides
Biochemistry
13
1186-1189
1974
Bacillus thermoproteolyticus
Titani, K.; Hermodson, M.A.; Ericsson, L.H.; Walsh, K.A.; Neurath, H.
Amino acid sequence of thermolysin. Isolation and characterization of the fragments obtained by cleavage with cyanogen bromide
Biochemistry
11
2427-2435
1972
Bacillus thermoproteolyticus
Nishino, N.; Powers, J.C.
Peptide hydroxamic acids as inhibitors of thermolysin
Biochemistry
17
2846-2850
1978
Bacillus thermoproteolyticus
Morgan, G.; Fruton, J.S.
Kinetics of the action of thermolysin on peptide substrates
Biochemistry
17
3562-3568
1978
Bacillus thermoproteolyticus
Rasnick, D.; Powers, J.C.
Active site directed irreversible inhibition of thermolysin
Biochemistry
17
4363-4369
1978
Bacillus thermoproteolyticus
Kam, C.M.; Nishino, N.; Powers, J.C.
Inhibition of thermolysin and carboxypeptidase A by phosphoramidates
Biochemistry
18
3032-3038
1979
Bacillus thermoproteolyticus
Nishino, N.; Powers, J.C.
Design of potent reversible inhibitors for thermolysin. Peptides containing zinc coordinating ligands and their use in affinity chromatography
Biochemistry
18
4340-4347
1979
Bacillus thermoproteolyticus
Holmes, M.A.; Matthews, B.W.
Binding of hydroxamic acid inhibitors to crystalline thermolysin suggests a pentacoordinate zinc intermediate in catalysis
Biochemistry
20
6912-6920
1981
Bacillus thermoproteolyticus
Weaver, L.H.; Lester, W.R.; Matthews, B.W.
A crystallographic study of the complex of phosphoramidon with thermolysin. A model for the presumed catalytic transition state and for the binding of extended substances
J. Mol. Biol.
114
119-132
1977
Bacillus thermoproteolyticus
Kester, W.R.; Matthews, B.W.
Crystallographic study of the binding of dipeptide inhibitors to thermolysin: implications for the mechanism of catalysis
Biochemistry
16
2506-2516
1977
Bacillus thermoproteolyticus
Tajima, M.; Urabe, I.; Yutani, K.; Okada, H.
Role of calcium ions in the thermostability of thermolysin and Bacillus subtilis var. amylosacchariticus neutral protease
Eur. J. Biochem.
64
243-247
1976
Bacillus thermoproteolyticus
Matthews, B.W.; Colman, P.M.; Jansonius, J.N.; Titani, K.; Walsh, K.A.; Neurath, H.
Structure of thermolysin
Nat. New Biol.
238
41-43
1972
Bacillus thermoproteolyticus
Hersh, L.B.; Morihara, K.
Comparison of the subsite specificity of the mammalian neutral endopeptidase 24.11 (enkephalinase) to the bacterial neutral endopeptidase thermolysin
J. Biol. Chem.
261
6433-6437
1986
Bacillus thermoproteolyticus
Fujiwara, K.; Tsuru, D.
Affinity chromatography of neutral and alkaline proteases from Bacillus subtilis and of thermolysin
J. Biochem.
76
883-886
1974
Bacillus thermoproteolyticus
Colman.P.M.; Jansonius, J.N.; Matthews, B.M.
The structure of thermolysin: an electron density map at 2-3 A resolution
J. Mol. Biol.
70
701-724
1972
Bacillus thermoproteolyticus
Morihara, K.; Tsuzuki, H.; Oka, T.
Comparison of the specificities of various neutral proteinases from microorganisms
Arch. Biochem. Biophys.
123
572-588
1968
Aspergillus oryzae, Bacillus thermoproteolyticus
Holland, D.R.; Tronrud, D.E.; Pley, H.W.; Flaherty, K.M.; Stark, W.; Jansonius, J.N.; McKay, D.B.; Matthews, B.W.
Structural comparison suggests that thermolysin and related neutral proteases undergo hinge-bending motion during catalysis
Biochemistry
31
11310-11316
1992
Bacillus thermoproteolyticus
Gaucher, J.F.; Selkti, M.; Prange, T.; Tomas, A.
The 2.2 A resolution structure of thermolysin (TLN) crystallized in the presence of potassium thiocyanate
Acta Crystallogr. Sect. D
58
2198-2200
2002
Bacillus thermoproteolyticus (P00800), Bacillus thermoproteolyticus
Marie-Claire, C.; Ruffet, E.; Antonczak, S.; Beaumont, A.; O'Donohue, M.; Roques, B.P.; Fournie-Zaluski, M.C.
Evidence by site-directed mutagenesis that arginine 203 of thermolysin and arginine 717 of neprilysin (neutral endopeptidase) play equivalent critical roles in substrate hydrolysis and inhibitor binding
Biochemistry
36
13938-13945
1997
Bacillus thermoproteolyticus, Bacillus thermoproteolyticus Rokko
Gaucher, J.F.; Selkti, M.; Tiraboschi, G.; Prange, T.; Roques, B.P.; Tomas, A.; Fournie-Zaluski, M.C.
Crystal structures of alpha-mercaptoacyldipeptides in the thermolysin active site: structural parameters for a Zn monodentation or bidentation in metalloendopeptidases
Biochemistry
38
12569-12576
1999
Bacillus thermoproteolyticus (P00800)
Ligne, T.; Pauthe, E.; Monti, J.P.; Gacel, G.; Larreta-Garde, V.
Additional data about thermolysin specificity in buffer- and glycerol-containing media
Biochim. Biophys. Acta
1337
143-148
1997
Bacillus thermoproteolyticus, Bacillus thermoproteolyticus Rokko
Kudryashova, E.V.; Mozhaev, V.V.; Balny, C.
Catalytic activity of thermolysin under extremes of pressure and temperature: modulation by metal ions
Biochim. Biophys. Acta
1386
199-210
1998
Bacillus thermoproteolyticus, Bacillus thermoproteolyticus Rokko
Takeuchi, T.; Bottcher, A.; Quezada, C.M.; Meade, T.J.; Gray, H.B.
Inhibition of thermolysin and human alpha-thrombin by cobalt(III) Schiff base complexes
Bioorg. Med. Chem.
7
815-819
1999
Bacillus thermoproteolyticus
Murakami, Y.; Chiba, K.; Oda, T.; Hirata, A.
Novel kinetic analysis of enzymatic dipeptide synthesis: effect of pH and substrates on thermolysin catalysis
Biotechnol. Bioeng.
74
406-415
2001
Bacillus thermoproteolyticus
Marie-Claire, C.; Ruffet, E.; Tiraboschi, G.; Fournie-Zaluski, M.C.
Differences in transition state stabilization between thermolysin (EC 3.4.24.27) and neprilysin (EC 3.4.24.11)
FEBS Lett.
438
215-219
1998
Bacillus thermoproteolyticus
Inouye, K.; Lee, S.B.; Nambu, K.; Tonomura, B.
Effects of pH, temperature, and alcohols on the remarkable activation of thermolysin by salts
J. Biochem.
122
358-364
1997
Bacillus thermoproteolyticus
Kuzuya, K.; Inouye, K.
Effects of cobalt-substitution of the active zinc ion in thermolysin on its activity and active-site microenvironment
J. Biochem.
130
783-788
2001
Bacillus thermoproteolyticus
Muta, Y.; Inouye, K.
Inhibitory effects of alcohols on thermolysin activity as examined using a fluorescent substrate
J. Biochem.
132
945-951
2002
Bacillus thermoproteolyticus
Baltora-Rosset, S.; Aboubeker, A.; Dupradeau, F.Y.; Pauthe, E.; Gacel, G.A.; Larreta-Garde, V.; Monti, J.P.
Structural studies by 1H NMR and molecular modeling of peptide substrates of thermolysin in relation with its proteasic activity in water and glycerol
J. Biomol. Struct. Dyn.
16
1061-1074
1999
Bacillus thermoproteolyticus, Bacillus thermoproteolyticus Rokko
English, A.C.; Done, S.H.; Caves, L.S.; Groom, C.R.; Hubbard, R.E.
Locating interaction sites on proteins: the crystal structure of thermolysin soaked in 2% to 100% isopropanol
Proteins
37
628-640
1999
Bacillus thermoproteolyticus
Kamo, M.; Inouye, K.; Nagata, K.; Tanokura, M.
Preliminary X-ray crystallographic analysis of thermolysin in the presence of 4 M NaCl
Acta crystallogr. Sect. D
61
710-712
2005
Bacillus thermoproteolyticus
Oneda, H.; Muta, Y.; Inouye, K.
Substrate-dependent activation of thermolysin by salt
Biosci. Biotechnol. Biochem.
68
1811-1813
2004
Bacillus thermoproteolyticus
Kawasaki, T.; Hoshino, Y.; Ishizu, Y.; Mizushiro, Y.; Okahata, Y.
Control of hydrolysis and condensation activities of thermolysin by ultrasound irradiation
Chem. Lett.
34
1602-1603
2005
Bacillus thermoproteolyticus
-
Oda, K.; Takahashi, T.; Takada, K.; Tsunemi, M.; Ng, K.K.; Hiraga, K.; Harada, S.
Exploring the subsite-structure of vimelysin and thermolysin using FRETS-libraries
FEBS Lett.
579
5013-5018
2005
Bacillus thermoproteolyticus
Lapointe, J.; Molle, D.; Gauthier, S.F.; Pouliot, Y.
Effect of calcium on thermolysin hydrolysis of ?-casein tryptic peptides
Int. Dairy J.
14
185-193
2004
Bacillus thermoproteolyticus
NNegue, M.; Miclo, L.; Girardet, J.; Campagna, S.; Molle, D.; Gaillard, J.
Proteolysis of bovine ?-lactalbumin by thermolysin during thermal denaturation
Int. Dairy J.
16
1157-1167
2006
Bacillus thermoproteolyticus
Matsumiya, Y.; Nishikawa, K.; Aoshima, H.; Inouye, K.; Kubo, M.
Analysis of autodegradation sites of thermolysin and enhancement of its thermostability by modifying Leu155 at an autodegradation site
J. Biochem.
135
547-553
2004
Bacillus thermoproteolyticus
Kusano, M.; Yasukawa, K.; Hashida, Y.; Inouye, K.
Engineering of the pH-dependence of thermolysin activity as examined by site-directed mutagenesis of Asn112 located at the active site of thermolysin
J. Biochem.
139
1017-1023
2006
Bacillus thermoproteolyticus (P00800)
Hernandez-Ledesma, B.; Ramos, M.; Recio, I.; Amigo, L.
Effect of beta-lactoglobulin hydrolysis with thermolysin under denaturing temperatures on the release of bioactive peptides
J. Chromatogr. A
1116
31-37
2006
Bacillus thermoproteolyticus
Matsumiya, Y.; Nishikawa, K.; Inouye, K.; Kubo, M.
Mutational effect for stability in a conserved region of thermolysin
Lett. Appl. Microbiol.
40
329-334
2005
Bacillus thermoproteolyticus
Inouye, K.; Minoda, M.; Takita, T.; Sakurama, H.; Hashida, Y.; Kusano, M.; Yasukawa, K.
Extracellular production of recombinant thermolysin expressed in Escherichia coli, and its purification and enzymatic characterization
Protein Expr. Purif.
46
248-255
2006
Bacillus thermoproteolyticus
Pedersen, N.R.; Kristensen, J.B.; Bauw, G.; Ravoo, B.J.; Darcy, R.; Larsen, K.L.; Pedersen, L.H.
Thermolysin catalyzes the synthesis of cyclodextrin esters in DMSO
Tetrahedron
16
615-622
2005
Bacillus thermoproteolyticus
-
Perez-Victoria, I.; Morales, J.C.
Regioselectivity in acylation of oligosaccharides catalyzed by the metalloprotease thermolysin
Tetrahedron
62
2361-2369
2006
Bacillus thermoproteolyticus
-
Yasukawa, K.; Inouye, K.
Improving the activity and stability of thermolysin by site-directed mutagenesis
Biochim. Biophys. Acta
1774
1281-1288
2007
Bacillus thermoproteolyticus
Takita, T.; Aono, T.; Sakurama, H.; Itoh, T.; Wada, T.; Minoda, M.; Yasukawa, K.; Inouye, K.
Effects of introducing negative charges into the molecular surface of thermolysin by site-directed mutagenesis on its activity and stability
Biochim. Biophys. Acta
1784
481-488
2008
Bacillus thermoproteolyticus (P00800)
Inouye, K.; Nakamura, K.; Kusano, M.; Yasukawa, K.
Improvement in performance of affinity gels containing Gly-D-Phe as a ligand to thermolysin due to increasing the spacer chain length
Biosci. Biotechnol. Biochem.
71
2083-2086
2007
Bacillus thermoproteolyticus
Inouye, K.; Kusano, M.; Hashida, Y.; Minoda, M.; Yasukawa, K.
Engineering, expression, purification, and production of recombinant thermolysin
Biotechnol. Annu. Rev.
13
43-64
2007
Geobacillus stearothermophilus, Bacillus thermoproteolyticus, Geobacillus stearothermophilus MK232
Tatsumi, C.; Hashida, Y.; Yasukawa, K.; Inouye, K.
Effects of site-directed mutagenesis of the surface residues Gln128 and Gln225 of thermolysin on its catalytic activity
J. Biochem.
141
835-842
2007
Bacillus thermoproteolyticus
Hashida, Y.; Inouye, K.
Kinetic analysis of the activation-and-inhibition dual effects of cobalt ion on thermolysin activity
J. Biochem.
141
843-853
2007
Bacillus thermoproteolyticus
Hashida, Y.; Inouye, K.
Molecular mechanism of the inhibitory effect of cobalt ion on thermolysin activity and the suppressive effect of calcium ion on the cobalt ion-dependent inactivation of thermolysin
J. Biochem.
141
879-888
2007
Bacillus thermoproteolyticus
Gokhale, N.H.; Bradford, S.; Cowan, J.A.
Stimulation and oxidative catalytic inactivation of thermolysin by copper.Cys-Gly-His-Lys
J. Biol. Inorg. Chem.
12
981-987
2007
Bacillus thermoproteolyticus
Pazhang, M.; Khajeh, K.; Ranjbar, B.; Hosseinkhani, S.
Effects of water-miscible solvents and polyhydroxy compounds on the structure and enzymatic activity of thermolysin
J. Biotechnol.
127
45-53
2006
Bacillus thermoproteolyticus
Blumberger, J.; Lamoureux, G.; Klein, M.L.
Peptide hydrolysis in thermolysin: ab initio QM/MM investigation of the Glu143-assisted water addition mechanism
J. Chem. Theory Comput.
3
1837-1850
2007
Bacillus thermoproteolyticus
Hu, Z.; Jiang, J.
Molecular dynamics simulations for water and ions in protein crystals
Langmuir
24
4215-4223
2008
Bacillus thermoproteolyticus
Owen, J.P.; Maddison, B.C.; Whitelam, G.C.; Gough, K.C.
Use of thermolysin in the diagnosis of prion diseases
Mol. Biotechnol.
35
161-170
2007
Bacillus thermoproteolyticus
Yasukawa, K.; Kusano, M.; Inouye, K.
A new method for the extracellular production of recombinant thermolysin by co-expressing the mature sequence and pro-sequence in Escherichia coli
Protein Eng. Des. Sel.
20
375-383
2007
Bacillus thermoproteolyticus
Yasukawa, K.; Kusano, M.; Nakamura, K.; Inouye, K.
Characterization of Gly-D-Phe, Gly-L-Leu, and D-Phe as affinity ligands to thermolysin
Protein Expr. Purif.
46
332-336
2006
Bacillus thermoproteolyticus
Cronier, S.; Gros, N.; Tattum, M.H.; Jackson, G.S.; Clarke, A.R.; Collinge, J.; Wadsworth, J.D.
Detection and characterization of proteinase K-sensitive disease-related prion protein with thermolysin
Biochem. J.
416
297-305
2008
Bacillus thermoproteolyticus (P00800)
Liu, Y.H.; Konermann, L.
Conformational dynamics of free and catalytically active thermolysin are indistinguishable by hydrogen/deuterium exchange mass spectrometry
Biochemistry
47
6342-6351
2008
Bacillus thermoproteolyticus
Kusano, M.; Yasukawa, K.; Inouye, K.
Insights into the catalytic roles of the polypeptide regions in the active site of thermolysin and generation of the thermolysin variants with high activity and stability
J. Biochem.
145
103-113
2009
Bacillus thermoproteolyticus
Khan, M.T.; Fuskevag, O.M.; Sylte, I.
Discovery of potent thermolysin inhibitors using structure based virtual screening and binding assays
J. Med. Chem.
52
48-61
2009
Bacillus thermoproteolyticus
Dong, M.; Liu, H.
Origins of the different metal preferences of Escherichia coli peptide deformylase and Bacillus thermoproteolyticus thermolysin: a comparative quantum mechanical/molecular mechanical study
J. Phys. Chem. B
112
10280-10290
2008
Bacillus thermoproteolyticus (P00800), Bacillus thermoproteolyticus
Marguerre, A.K.; Kraemer, R.
Lanthanide-based fluorogenic peptide substrate for the highly sensitive detection of thermolysin
Bioorg. Med. Chem. Lett.
19
5757-5759
2009
Bacillus thermoproteolyticus
Khan, M.T.; Khan, R.; Wuxiuer, Y.; Arfan, M.; Ahmed, M.; Sylte, I.
Identification of novel quinazolin-4(3H)-ones as inhibitors of thermolysin, the prototype of the M4 family of proteinases
Bioorg. Med. Chem.
18
4317-4327
2010
Bacillus thermoproteolyticus
Liu, Z.; Zhang, P.; Zhou, Y.; Qin, H.; Shen, T.
Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3
Braz. J. Med. Biol. Res.
43
451-459
2010
Bacillus thermoproteolyticus
Adekoya, O.A.; Sylte, I.
The thermolysin family (M4) of enzymes: therapeutic and biotechnological potential
Chem. Biol. Drug Des.
73
7-16
2009
Bacillus cereus, Bacillus thermoproteolyticus
Englert, L.; Silber, K.; Steuber, H.; Brass, S.; Over, B.; Gerber, H.D.; Heine, A.; Diederich, W.E.; Klebe, G.
Fragment-based lead discovery: screening and optimizing fragments for thermolysin inhibition
ChemMedChem
5
930-940
2010
Bacillus thermoproteolyticus (P00800)
Asaoka, K.; Yasukawa, K.; Inouye, K.
Coagulation of soy proteins induced by thermolysin and comparison of the coagulation reaction with that induced by subtilisin Carlsberg
Enzyme Microb. Technol.
44
229-234
2009
Bacillus thermoproteolyticus
-
Kusano, M.; Yasukawa, K.; Inouye, K.
Synthesis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester catalyzed by thermolysin variants with improved activity
Enzyme Microb. Technol.
46
320-325
2010
Bacillus thermoproteolyticus
-
Kusano, M.; Yasukawa, K.; Inouye, K.
Effects of the mutational combinations on the activity and stability of thermolysin
J. Biotechnol.
147
7-16
2010
Bacillus thermoproteolyticus
Hu, Z.; Jiang, J.
Chiral separation of racemic phenylglycines in thermolysin crystal: a molecular simulation study
J. Phys. Chem. B
113
15851-15857
2009
Bacillus thermoproteolyticus
Ceruso, M.; Howe, N.; Malthouse, J.P.
Mechanism of the binding of Z-L-tryptophan and Z-L-phenylalanine to thermolysin and stromelysin-1 in aqueous solutions
Biochim. Biophys. Acta
1824
303-310
2012
Bacillus sp. (in: Bacteria), Bacillus thermoproteolyticus
Menach, E.; Yasukawa, K.; Inouye, K.
Effects of site-directed mutagenesis of the loop residue of the N-terminal domain Gly117 of thermolysin on its catalytic activity
Biosci. Biotechnol. Biochem.
74
2457-2462
2010
Bacillus thermoproteolyticus
Birrane, G.; Bhyravbhatla, B.; Navia, M.
Synthesis of aspartame by thermolysin: an x-ray structural study
ACS Med. Chem. Lett.
5
706-710
2014
Bacillus thermoproteolyticus (P00800)
Chen, F.; Zhang, F.; Du, F.; Wang, A.; Gao, W.; Wang, Q.; Yin, X.; Xie, T.
A novel and efficient method for the immobilization of thermolysin using sodium chloride salting-in and consecutive microwave irradiation
Biores. Technol.
115
158-163
2012
Bacillus thermoproteolyticus, Bacillus thermoproteolyticus Rokko
Sato, Y.; Toyoda, T.; Shimizu-Ibuka, A.; Tamura, T.; Kobayashi-Hattori, K.; Nakamura, T.; Arai, S.; Mura, K.
Novel angiotensin I-converting enzyme inhibitory peptides found in a thermolysin-treated elastin with antihypertensive activity
Biosci. Biotechnol. Biochem.
76
1329-1333
2012
Bacillus thermoproteolyticus, Bacillus thermoproteolyticus Rokko
Adekoya, O.; Sjoeli, S.; Wuxiuer, Y.; Bilto, I.; Marques, S.; Santos, M.; Nuti, E.; Cercignani, G.; Rossello, A.; Winberg, J.; Sylte, I.
Inhibition of pseudolysin and thermolysin by hydroxamate-based MMP inhibitors
Eur. J. Med. Chem.
89
340-348
2014
Bacillus thermoproteolyticus
Hardre, H.; Kuhn, L.; Albrieux, C.; Jouhet, J.; Michaud, M.; Seigneurin-Berny, D.; Falconet, D.; Block, M.; Marechal, E.
The selective biotin tagging and thermolysin proteolysis of chloroplast outer envelope proteins reveals information on protein topology and association into complexes
Front. Plant Sci.
5
203
2014
Bacillus thermoproteolyticus
Guinane, C.; Kent, R.; Norberg, S.; O'Connor, P.; Cotter, P.; Hill, C.; Fitzgerald, G.; Stanton, C.; Ross, R.
Generation of the antimicrobial peptide caseicin A from casein by hydrolysis with thermolysin enzymes
Int. Dairy J.
49
1-7
2015
Bacillus thermoproteolyticus
-
Menach, E.; Yasukawa, K.; Inouye, K.
Effects of site-directed mutagenesis of Asn116 in the beta-hairpin of the N-terminal domain of thermolysin on its activity and stability
J. Biochem.
152
231-239
2012
Bacillus thermoproteolyticus (P00800)
Kim, C.; Lee, D.; Lee, C.; Ahn, I.
Dityrosine-based substrates for the selective and sensitive assay of thermolysin
J. Ind. Eng. Chem.
21
248-253
2015
Bacillus thermoproteolyticus
-
Osago, H.; Kobayashi-Miura, M.; Hamasaki, Y.; Hara, N.; Hiyoshi, M.; Tsuchiya, M.
Complete solubilization of cartilage using the heat-stable protease thermolysin for comprehensive GAG analysis
Anal. Biochem.
548
115-118
2018
Bacillus thermoproteolyticus
Rocha-Martin, J.; Fernandez-Lorente, G.; Guisan, J.
Sequential hydrolysis of commercial casein hydrolysate by immobilized trypsin and thermolysin to produce bioactive phosphopeptides
Biocatal. Biotransform.
36
159-171
2018
Bacillus thermoproteolyticus (P00800)
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