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Literature summary extracted from
- The effect of disulfide bond introduction and related Cys/Ser mutations on the stability of a cyclohexanone monooxygenase (2015), J. Biotechnol., 214, 199-211 .
Application
| EC Number | Application | Comment | Organism |
|---|---|---|---|
| 1.14.13.22 | synthesis | cyclohexanone monooxygenase (CHMO) from Acinetobacter sp. NCIMB 9871 is a prototype Baeyer-Villiger monooxygenases (BVMO). The enzyme shows an impressive substrate scope with a high chemo-, regio- and/or enantioselectivity. BVMO reactions are often difficult, if not impossible to achieve by chemical approaches and this makes these enzymes highly desired candidates for industrial applications. The industrial use is hampered by several factors related to the lack of stability of these biocatalysts. An easy computational method is used for the prediction of stabilizing disulfide bonds in the cyclohexanone monooxygenase-scaffold. The most promising predicted disulfide pairs are created and biochemically characterized. The T415C single point variant is the most stable variant with a 30fold increased long-term stability (33% residual activity after 24 h incubation at 25°C) | Acinetobacter johnsonii |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.14.13.22 | expression in Escherichia coli | Acinetobacter johnsonii |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.14.13.22 | T415C | most stable variant with a 30fold increased long-term stability (33% residual activity after 24 h incubation at 25°C), the melting temperature of the variant is increased by 6°C | Acinetobacter johnsonii |
| 1.14.13.22 | T415C/A463C | the melting temperature of the variant is increased by 5°C with simultaneous improved long-term stability | Acinetobacter johnsonii |
| 1.14.13.22 | Y411C/A463C | the most oxidatively stable variant Y411C-A463C retains nearly 60% activity after incubation with 25 mM H2O2 whereas the wild type retains only 16% | Acinetobacter johnsonii |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.13.22 | 0.003 | - |
cyclohexanone | 25°C, pH 8.0, mutant enzyme P254C/D290C | Acinetobacter johnsonii |  |
| 1.14.13.22 | 0.0046 | - |
cyclohexanone | 25°C, pH 8.0, wild-type enzyme | Acinetobacter johnsonii | |
| 1.14.13.22 | 0.0109 | - |
cyclohexanone | 25°C, pH 8.0, mutant enzyme T415C/A463C | Acinetobacter johnsonii | |
| 1.14.13.22 | 0.0138 | - |
cyclohexanone | 25°C, pH 8.0, mutant enzyme Y411C/A463C | Acinetobacter johnsonii | |
| 1.14.13.22 | 0.0209 | - |
cyclohexanone | 25°C, pH 8.0, mutant enzyme T415C | Acinetobacter johnsonii | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.13.22 | Acinetobacter johnsonii | Q9R2F5 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.13.22 | - |
Acinetobacter johnsonii |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.13.22 | 4-methylcyclohexanone + NADPH + H+ + O2 | - |
Acinetobacter johnsonii | ? | - |
? | |
| 1.14.13.22 | cyclohexanone + NADPH + H+ + O2 | - |
Acinetobacter johnsonii | hexano-6-lactone + NADP+ + H2O | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.14.13.22 | CHMO | prototype Baeyer-Villiger monooxygenases (BVMO) | Acinetobacter johnsonii |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.22 | 25 | - |
mutant T415C is the most stable variant with a 30fold increased long-term stability, 33% residual activity after 24 h incubation at 25°C | Acinetobacter johnsonii |
| 1.14.13.22 | 31.6 | - |
Tm-value of wilde-type enzyme | Acinetobacter johnsonii |
| 1.14.13.22 | 36.4 | - |
Tm-value of mutant enzyme T415C/A463C | Acinetobacter johnsonii |
| 1.14.13.22 | 37.5 | - |
Tm-value of mutant enzyme T415C | Acinetobacter johnsonii |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.13.22 | 6.1 | - |
cyclohexanone | 25°C, pH 8.0, mutant enzyme P254C/D290C | Acinetobacter johnsonii | |
| 1.14.13.22 | 6.15 | - |
cyclohexanone | 25°C, pH 8.0, mutant enzyme Y411C/A463C | Acinetobacter johnsonii | |
| 1.14.13.22 | 38.5 | - |
cyclohexanone | 25°C, pH 8.0, wild-type enzyme | Acinetobacter johnsonii | |
| 1.14.13.22 | 44.9 | - |
cyclohexanone | 25°C, pH 8.0, mutant enzyme T415C/A463C | Acinetobacter johnsonii | |
| 1.14.13.22 | 57.4 | - |
cyclohexanone | 25°C, pH 8.0, mutant enzyme T415C | Acinetobacter johnsonii | |
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