Application | Comment | Organism |
---|---|---|
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 | Bacillus thermoproteolyticus |
Protein Variants | Comment | Organism |
---|---|---|
D150A | mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type | Bacillus thermoproteolyticus |
D150E | mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type | Bacillus thermoproteolyticus |
D150W | mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type | Bacillus thermoproteolyticus |
F114A | mutant with reduced activity | Bacillus thermoproteolyticus |
I168A | mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type | Bacillus thermoproteolyticus |
N227H | mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type | Bacillus thermoproteolyticus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.7 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
0.8 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
1.3 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
2.1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
2.1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
5.8 | - |
L-phenylalanine methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
7 | - |
L-phenylalanine methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
7.3 | - |
L-phenylalanine methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
8.7 | - |
L-phenylalanine methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
8.8 | - |
L-phenylalanine methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
11.3 | - |
L-phenylalanine methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
12.9 | - |
L-phenylalanine methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
49 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
54 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
57 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
60 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
62 | - |
N-carbobenzoxy-L-aspartic acid | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
70 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
105 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus thermoproteolyticus | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-carbobenzoxy-L-Asp-L-Phe-methyl ester + H2O | - |
Bacillus thermoproteolyticus | N-carbobenzoxy-L-aspartic acid + L-phenylalanine methyl ester | - |
r | |
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide + H2O | - |
Bacillus thermoproteolyticus | ? | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.3 | - |
L-phenylalanine methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
0.3 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
1.3 | - |
L-phenylalanine methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
1.3 | - |
N-carbobenzoxy-L-aspartic acid | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
1.5 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
2.1 | - |
L-phenylalanine methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
2.1 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
2.3 | - |
L-phenylalanine methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
2.3 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
2.5 | - |
L-phenylalanine methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
2.5 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
3.2 | - |
L-phenylalanine methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
3.2 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
4.9 | - |
L-phenylalanine methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
4.9 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
9.5 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
12.3 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
17.1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
20.5 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
24.4 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
49.4 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
around pH 7.0 | Bacillus thermoproteolyticus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.5 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
9.3 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
11.7 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
13.7 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
16 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
23 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
27.6 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus |