3.4.24.27: thermolysin
This is an abbreviated version!
For detailed information about thermolysin, go to the full flat file.
Word Map on EC 3.4.24.27
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3.4.24.27
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chymotrypsin
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elastase
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metalloprotease
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subtilisin
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staphylococcus
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edman
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pepsin
-
aureus
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carboxypeptidase
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endopeptidase
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bromide
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cyanogen
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collagenase
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proteinases
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dipeptide
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angiotensin
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pronase
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metalloproteinases
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alpha-chymotrypsin
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thermolytic
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hydrolysates
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metalloendopeptidase
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stearothermophilus
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endoproteinase
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phosphoramidon
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i-converting
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enkephalinase
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rhodopsin
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2-macroglobulin
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3.4.24.11
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cell-binding
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alcalase
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ace-inhibitory
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hexxh
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dispase
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aspartame
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thiorphan
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neprilysin
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s-carboxymethylated
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half-cystine
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astacin
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synthesis
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industry
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nutrition
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food industry
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diagnostics
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medicine
-
analysis
- 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
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3.4.24.11
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cell-binding
- alcalase
-
ace-inhibitory
-
hexxh
-
dispase
- aspartame
- thiorphan
- neprilysin
-
s-carboxymethylated
-
half-cystine
- astacin
- synthesis
- industry
- nutrition
- food industry
- diagnostics
- medicine
- analysis
Reaction
preferential cleavage: -/-Leu > -/-Phe =
Synonyms
Bacillus thermoproteolyticus neutral proteinase, EC 3.4.24.4, hspA, LIC13322, Neutral metalloproteinase, NprM, protease type X, proteinase type X, Proteinase, Bacillus thermoproteolyticus neutral, protex 14L, Thermoase, thermoase PC10F, Thermoase Y10, thermolysin, thermolysin-like protease, Thermostable neutral proteinase, TL, TLN, TLP, TLP-ste
ECTree
3.4.24.27 thermolysinAdvanced search results
results (
results (Engineering
Engineering on EC 3.4.24.27 - thermolysin
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
L155A
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site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is slightly enhanced compared to the wild-type enzyme
L155A/I156N
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site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme
L155A/I156V
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site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme
L155S
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site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is slightly enhanced compared to the wild-type enzyme
L155S/I156N
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site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme
L155S/I156V
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site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme
A4T/G8C/T56A/G58A/N60C/T63F/S65P/A69P
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the mutant shows altered thermodynamics
A4T/T56A/G58A/T63F/S65P/A69P
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the mutant shows altered thermodynamics
D150A
D150E
D150H
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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
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51% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
D150R
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44% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
D150W
D170A
DELTA127
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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
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site-directed mutagenesis, E143A might exist as a complex with the propetide in the supernatant, inactive mutant, the autocatalytic activity is affected
F114A
F114H
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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
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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
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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
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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
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the mutant shows 500fold decreased catalytic efficiency compared to the wild-type enzyme
I168A
I168H
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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
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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
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the triple mutant shows improved activity and stability with about 30% casein-hydrolytic activity compared to the wild type enzyme
L144S/D150W/N227H
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the mutant shows 10fold decreased catalytic efficiency compared to the wild-type enzyme
L144S/I168A
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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
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the mutant shows about 80% casein-hydrolytic activity compared to the wild type enzyme
L155F
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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
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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
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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
N112D
N112E
N112H
N112K
N112R
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
N116D/Q119R/D150Q/Q225R
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the mutant shows 4fold decreased catalytic efficiency 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
N227A
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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
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11% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
N227E
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36% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
N227H
N227K
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29% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
N227R
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55% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
P208H
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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
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site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
Q128E
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site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme
Q128K
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Q225A
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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
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site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
Q225E
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site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme
Q225K
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site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
Q225R
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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
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the mutant shows 5fold decreased catalytic efficiency compared to the wild-type enzyme
R203M
S103A
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the mutant shows 3fold decreased catalytic efficiency compared to the wild-type enzyme
S169A
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112% residual activity with casein, 64% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
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
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
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5% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
S234E
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4% residual activity with casein, 7% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
S234H
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32% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
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
S53D/G8C/N60C/S65P
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the mutant shows about 110% casein-hydrolytic activity compared to the wild type enzyme
S53D/L155A
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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
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the mutant shows about 70% casein-hydrolytic activity compared to the wild type enzyme
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
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
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6% residual activity with casein, 12% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157A
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11% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157D
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1% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157E
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13% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157H
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24% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide
Y157K
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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
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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
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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
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the triple mutation increases the stability of thermolysin as high as the individual mutations do
G8C/N60C/S65P/L144S
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site-directed mutagenesis, the mutant shows about 6fold increased catalytic effiency compared to the wild-type enzyme
G8C/N60C/S65P/L144S
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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
N112A
no enzymic activity in supernatant of cells expressing mutant
N112A
-
site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected
N112D
supernatants of cells expressing mutant show 18% of wild-type activity
N112D
-
site-directed mutagenesis, the autocatalytic activity is affected
N112E
supernatants of cells expressing mutant show 5% of wild-type activity
N112E
-
site-directed mutagenesis, the autocatalytic activity is affected
N112H
no enzymic activity in supernatant of cells expressing mutant
N112H
-
site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected
N112K
no enzymic activity in supernatant of cells expressing mutant
N112K
-
site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected
N112R
no enzymic activity in supernatant of cells expressing mutant
N112R
-
site-directed mutagenesis, inactive mutant, 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
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
additional information
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a mutant thermolysin is affected by its autocatalytic digestion activity
additional information
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generation of an engineered enzyme with a higher activity in the synthesis of N-carbobenzyloxy L-Asp-L-Phe methyl ester
additional information
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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
additional information
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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
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additional information
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generation of an engineered enzyme with a higher activity in the synthesis of N-carbobenzyloxy L-Asp-L-Phe methyl ester
additional information
-
generation of an engineered enzyme with a higher activity in the synthesis of N-carbobenzyloxy L-Asp-L-Phe methyl ester
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