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Literature summary extracted from
- Directed evolution of phenylacetone monooxygenase as an active catalyst for the Baeyer-Villiger conversion of cyclohexanone to caprolactone (2015), Biotechnol. Bioeng., 112, 1354-1364 .
Application
| EC Number | Application | Comment | Organism |
|---|---|---|---|
| 1.14.13.92 | synthesis | phenylacetone monooxygenase (PAMO) is an exceptionally robust Baeyer-Villiger monooxygenase, which makes it ideal for potential industrial applications, usage as an active catalyst for the Baeyer-Villiger conversion of cyclohexanone to caprolactone, which is important as monomer in polymer science | Thermobifida fusca |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.14.13.92 | A442P | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 81% conversion rate | Thermobifida fusca |
| 1.14.13.92 | A442P/ L443I/S444Q | random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 43% conversion rate | Thermobifida fusca |
| 1.14.13.92 | A442P/ L443V/S444Q | random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 45% conversion rate | Thermobifida fusca |
| 1.14.13.92 | A442P/L443I | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 45% conversion rate | Thermobifida fusca |
| 1.14.13.92 | A442P/L443L/S444Q | random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 41% conversion rate | Thermobifida fusca |
| 1.14.13.92 | A442P/L443T/S444Q | random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 56% conversion rate | Thermobifida fusca |
| 1.14.13.92 | A442P/L443V | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 90% conversion rate | Thermobifida fusca |
| 1.14.13.92 | A442P/L443W | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 74% conversion rate | Thermobifida fusca |
| 1.14.13.92 | A442P/L443W/ S444Q | random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 33% conversion rate | Thermobifida fusca |
| 1.14.13.92 | L443V | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 53% conversion rate | Thermobifida fusca |
| 1.14.13.92 | L443V/S444M | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 53% conversion rate | Thermobifida fusca |
| 1.14.13.92 | L443V/S444Q | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 40-45% conversion rate | Thermobifida fusca |
| 1.14.13.92 | L443V/S444Q | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 59% conversion rate | Thermobifida fusca |
| 1.14.13.92 | L443V/S444T | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 57% conversion rate | Thermobifida fusca |
| 1.14.13.92 | additional information | directed evolution of phenylacetone monooxygenase as an active catalyst for the Baeyer-Villiger conversion of cyclohexanone to caprolactone using iterative saturation mutagenesis, mutant screening, overview. Molecular dynamics simulations and induced fit docking of wild-type and mutant enzymes with cyclohexanone. The mutants are used in the whole cell system of Escherichia coli cells | Thermobifida fusca |
| 1.14.13.92 | Q93N/P94D/P440F | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at low rate | Thermobifida fusca |
| 1.14.13.92 | S441D/A442E | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 73% conversion rate | Thermobifida fusca |
| 1.14.13.92 | S441G/A442P/L443T/S444Q | site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at about 90% conversion rate | Thermobifida fusca |
| 1.14.13.92 | S441G/A442T | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 48% conversion rate | Thermobifida fusca |
| 1.14.13.92 | S441H | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 34% conversion rate | Thermobifida fusca |
| 1.14.13.92 | S441H/A442P | random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 78% conversion rate | Thermobifida fusca |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.13.92 | 0.266 | - |
cyclohexanone | pH 8.0, 25°C, recombinant mutant S441G/A442P/L443T/S444Q | Thermobifida fusca |  |
| 1.14.13.92 | 0.698 | - |
cyclohexanone | pH 8.0, 25°C, recombinant mutant A442P/L443V | Thermobifida fusca | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.13.92 | phenylacetone + NADPH + H+ + O2 | Thermobifida fusca | - |
benzyl acetate + NADP+ + H2O | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.13.92 | Thermobifida fusca | Q47PU3 | - |
- |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.13.92 | cyclohexanone + NADPH + H+ + O2 | substrate of enzyme mutants, not of wild-type, overview | Thermobifida fusca | epsilon-caprolactone + NADP+ + H2O | - |
? | |
| 1.14.13.92 | phenylacetone + NADPH + H+ + O2 | - |
Thermobifida fusca | benzyl acetate + NADP+ + H2O | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.14.13.92 | PAMO | - |
Thermobifida fusca |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.92 | 25 | - |
in vitro assay at | Thermobifida fusca |
| 1.14.13.92 | 30 | 37 | in vivo assay at | Thermobifida fusca |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.92 | 57.5 | - |
Tm value for enzyme PAMO mutant A442P/L443V | Thermobifida fusca |
| 1.14.13.92 | 58.5 | - |
Tm value for enzyme PAMO mutant S441G/A442P/L443T/S444Q | Thermobifida fusca |
| 1.14.13.92 | 60.5 | - |
Tm value for wild-type enzyme PAMO | Thermobifida fusca |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.13.92 | 0.156 | - |
cyclohexanone | pH 8.0, 25°C, recombinant mutant S441G/A442P/L443T/S444Q | Thermobifida fusca | |
| 1.14.13.92 | 0.304 | - |
cyclohexanone | pH 8.0, 25°C, recombinant mutant A442P/L443V | Thermobifida fusca | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.92 | 8 | - |
in vitro assay at | Thermobifida fusca |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.13.92 | NADPH | - |
Thermobifida fusca | |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 1.14.13.92 | additional information | molecular dynamics simulations and induced fit docking of wild-type and mutant enzymes with cyclohexanone | Thermobifida fusca |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.13.92 | 0.436 | - |
cyclohexanone | pH 8.0, 25°C, recombinant mutant A442P/L443V | Thermobifida fusca | |
| 1.14.13.92 | 0.577 | - |
cyclohexanone | pH 8.0, 25°C, recombinant mutant S441G/A442P/L443T/S444Q | Thermobifida fusca | |
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