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Literature summary extracted from
- Synthesis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester catalyzed by thermolysin variants with improved activity (2010), Enzyme Microb. Technol., 46, 320-325.
No PubMed abstract available
Application
| EC Number | Application | Comment | Organism |
|---|---|---|---|
| 3.4.24.27 | 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
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.4.24.27 | 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 |
| 3.4.24.27 | 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 |
| 3.4.24.27 | 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 |
| 3.4.24.27 | F114A | mutant with reduced activity | Bacillus thermoproteolyticus |
| 3.4.24.27 | 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 |
| 3.4.24.27 | 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]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.4.24.27 | 0.7 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus |  |
| 3.4.24.27 | 0.8 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 1.3 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 2.1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 2.1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 5.8 | - |
L-phenylalanine methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 7 | - |
L-phenylalanine methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 7.3 | - |
L-phenylalanine methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 8.7 | - |
L-phenylalanine methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 8.8 | - |
L-phenylalanine methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 11.3 | - |
L-phenylalanine methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 12.9 | - |
L-phenylalanine methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 49 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 54 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 57 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 60 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 62 | - |
N-carbobenzoxy-L-aspartic acid | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 70 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 105 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.4.24.27 | Bacillus thermoproteolyticus | - |
- |
- |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.4.24.27 | N-carbobenzoxy-L-Asp-L-Phe-methyl ester + H2O | - |
Bacillus thermoproteolyticus | N-carbobenzoxy-L-aspartic acid + L-phenylalanine methyl ester | - |
r | |
| 3.4.24.27 | N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide + H2O | - |
Bacillus thermoproteolyticus | ? | - |
? | |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.4.24.27 | 0.3 | - |
L-phenylalanine methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 0.3 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 1.3 | - |
L-phenylalanine methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 1.3 | - |
N-carbobenzoxy-L-aspartic acid | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 1.5 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 2.1 | - |
L-phenylalanine methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 2.1 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 2.3 | - |
L-phenylalanine methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 2.3 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 2.5 | - |
L-phenylalanine methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 2.5 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 3.2 | - |
L-phenylalanine methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 3.2 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 4.9 | - |
L-phenylalanine methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 4.9 | - |
N-carbobenzoxy-L-aspartic acid | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 9.5 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 12.3 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 17.1 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 20.5 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 24.4 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 49.4 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.4.24.27 | 7 | - |
around pH 7.0 | Bacillus thermoproteolyticus |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.4.24.27 | 1.5 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme F114A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 9.3 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | wild type enzyme, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 11.7 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 13.7 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme N227H, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 16 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme I168A, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 23 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150W, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
| 3.4.24.27 | 27.6 | - |
N-carbobenzoxy-L-Asp-L-Phe-methyl ester | mutant enzyme D150E, at pH 7.5, at 25°C | Bacillus thermoproteolyticus | |
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