Application | Comment | Organism |
---|---|---|
analysis | application of an aerobic, high-throughput growth selection platform in Escherichia coli for the discovery of thermostability enhancing mutations for CHMO. The selection employs growth for the easy readout of CHMO activity in vivo, by requiring nicotinamide adenine dinucleotide phosphate (NADPH)-consuming enzymes to restore cellular redox balance | Acinetobacter sp. |
Protein Variants | Comment | Organism |
---|---|---|
A245G/A288V | variant retains native activity, exhibited about 4.4fold improvement in residual activity after 30°C incubation, and demonstrates about 5fold higher cyclohexanone conversion at 37°C compared to the wild-type | Acinetobacter sp. |
A245G/A288V/T415C | variant shows improved thermostability at 45°C | Acinetobacter sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0041 | - |
cyclohexanone | wild-type, pH 7.7, 25°C | Acinetobacter sp. | |
0.0056 | - |
cyclohexanone | mutant A245G/A288V, pH 7.7, 25°C | Acinetobacter sp. | |
0.0366 | - |
NADPH | wild-type, pH 7.7, 25°C | Acinetobacter sp. | |
0.0433 | - |
NADPH | mutant A245G/A288V, pH 7.7, 25°C | Acinetobacter sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acinetobacter sp. | P12015 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cyclohexanone + NADPH + H+ + O2 | - |
Acinetobacter sp. | hexano-6-lactone + NADP+ + H2O | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
9.67 | - |
NADPH | mutant A245G/A288V, pH 7.7, 25°C | Acinetobacter sp. | |
11.6 | - |
cyclohexanone | mutant A245G/A288V, pH 7.7, 25°C | Acinetobacter sp. | |
12.9 | - |
cyclohexanone | wild-type, pH 7.7, 25°C | Acinetobacter sp. | |
13.5 | - |
NADPH | wild-type, pH 7.7, 25°C | Acinetobacter sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
220 | - |
NADPH | mutant A245G/A288V, pH 7.7, 25°C | Acinetobacter sp. | |
370 | - |
NADPH | wild-type, pH 7.7, 25°C | Acinetobacter sp. | |
2050 | - |
cyclohexanone | mutant A245G/A288V, pH 7.7, 25°C | Acinetobacter sp. | |
3150 | - |
cyclohexanone | wild-type, pH 7.7, 25°C | Acinetobacter sp. |