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Literature summary for 3.1.1.3 extracted from
- Enhancing the thermostability of Rhizomucor miehei lipase with a limited screening library by rational-design point mutations and disulfide bonds (2018), Appl. Environ. Microbiol., 84, e02129-17 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli | Rhizomucor miehei |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| T18K/T22I/E230I | thermostable mutant, melting temperature is 10 degrees higher than wild-type | Rhizomucor miehei |
| T18K/T22I/S56C/N63C/V189C/E230I/D238C | thermostable mutant, residues S56C/N63C and V189C/D238C may form additional disulfide bonds. Melting temperature is 14.3 degrees higher than wild-type and half-life at 70°C is 12.5fold increased | Rhizomucor miehei |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.17 | - |
4-nitrophenyl octanoate | wild-type, pH 8.0, 45°C | Rhizomucor miehei |  |
| 1.53 | - |
4-nitrophenyl octanoate | mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C, pH 8.0, 60°C | Rhizomucor miehei | |
| 2.01 | - |
4-nitrophenyl octanoate | mutant T18K/T22I/E230I, pH 8.0, 55°C | Rhizomucor miehei | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Rhizomucor miehei | A0A2D3HJ61 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl octanoate + H2O | - |
Rhizomucor miehei | 4-nitrophenol + octanoate | - |
? | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 45 | - |
wild-type | Rhizomucor miehei |
| 55 | - |
mutant T18K/T22I/E230I | Rhizomucor miehei |
| 60 | - |
mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C | Rhizomucor miehei |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 58.7 | - |
melting temperature, wild-type | Rhizomucor miehei |
| 65 | - |
120 min, 13% residual activity for wild-type, 95% for mutant T18K/T22I/E230I | Rhizomucor miehei |
| 68.1 | - |
melting temperature, mutant T18K/T22I/E230I | Rhizomucor miehei |
| 70 | - |
wild-type, half-life 2.2 min, mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C, half-life 27.5 min | Rhizomucor miehei |
| 73 | - |
melting temperature, mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C | Rhizomucor miehei |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 24.05 | - |
4-nitrophenyl octanoate | wild-type, pH 8.0, 45°C | Rhizomucor miehei | |
| 42.34 | - |
4-nitrophenyl octanoate | mutant T18K/T22I/E230I, pH 8.0, 55°C | Rhizomucor miehei | |
| 43.71 | - |
4-nitrophenyl octanoate | mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C, pH 8.0, 60°C | Rhizomucor miehei | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
wild-type | Rhizomucor miehei |
| 8.5 | - |
mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C | Rhizomucor miehei |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 20.6 | - |
4-nitrophenyl octanoate | wild-type, pH 8.0, 45°C | Rhizomucor miehei | |
| 21.1 | - |
4-nitrophenyl octanoate | mutant T18K/T22I/E230I, pH 8.0, 55°C | Rhizomucor miehei | |
| 28.6 | - |
4-nitrophenyl octanoate | mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C, pH 8.0, 60°C | Rhizomucor miehei | |
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