Application | Comment | Organism |
---|---|---|
synthesis | application of directed evolution using iterative saturation mutagenesis as a means to engineer LEH mutants showing broad substrate scope with high stereoselectivity. Mutants are obtained which catalyze the desymmetrization of cyclopentene-oxide with stereoselective formation of either the (R,R)- or the (S,S)-diol on an optional basis. The mutants prove to be excellent catalysts for the desymmetrization of other meso-epoxides and for the hydrolytic kinetic resolution of racemic substrates | Rhodococcus erythropolis |
Protein Variants | Comment | Organism |
---|---|---|
L114C/I116V | substrate cyclopentene-oxide, 72% conversion, (S,S)-product with 68% enantiomeric excess | Rhodococcus erythropolis |
L114I/I116V | substrate cyclopentene-oxide, 74% conversion, (S,S)-product with 50% enantiomeric excess | Rhodococcus erythropolis |
L114V/I116V | substrate cyclopentene-oxide, 72% conversion, (S,S)-product with 60% enantiomeric excess | Rhodococcus erythropolis |
L74I/I80C | substrate cyclopentene-oxide, 75% conversion, (R,R)-product with 66% enantiomeric excess | Rhodococcus erythropolis |
L74I/I80V | substrate cyclopentene-oxide, 75% conversion, (R,R)-product with 58% enantiomeric excess | Rhodococcus erythropolis |
L74V/I80V | substrate cyclopentene-oxide, 67% conversion, (R,R)-product with 53% enantiomeric excess | Rhodococcus erythropolis |
M32C/I80F/L114C/I116V | substrate rac-2-(phenoxymethyl)oxirane, 31% conversion, (2R)-product with 92% enantiomeric excess. Substrate rac-1-methyl-7-oxabicyclo[4.1.0]heptane, 99% conversion, (1S,2S)-product with 55% enantiomeric excess | Rhodococcus erythropolis |
M32L/L35C | substrate cyclopentene-oxide, 78% conversion, (S,S)-product with 16% enantiomeric excess | Rhodococcus erythropolis |
M32L/L35F | substrate cyclopentene-oxide, 79% conversion, (S,S)-product with 24% enantiomeric excess | Rhodococcus erythropolis |
M32L/L35V | substrate cyclopentene-oxide, 78% conversion, (S,S)-product with 10% enantiomeric excess | Rhodococcus erythropolis |
M78F/V83I | substrate cyclopentene-oxide, 82% conversion, (R,R)-product with 29% enantiomeric excess | Rhodococcus erythropolis |
M78I/V83I | substrate cyclopentene-oxide, 80% conversion, (R,R)-product with 13% enantiomeric excess | Rhodococcus erythropolis |
M78V/V83I | substrate cyclopentene-oxide, 68% conversion, (R,R)-product with 7% enantiomeric excess | Rhodococcus erythropolis |
additional information | application of directed evolution using iterative saturation mutagenesis as a means to engineer LEH mutants showing broad substrate scope with high stereoselectivity. Mutants are obtained which catalyze the desymmetrization of cyclopentene-oxide with stereoselective formation of either the (R,R)- or the (S,S)-diol on an optional basis. The mutants prove to be excellent catalysts for the desymmetrization of other meso-epoxides and for the hydrolytic kinetic resolution of racemic substrates | Rhodococcus erythropolis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus erythropolis | - |
- |
- |
Rhodococcus erythropolis DCL 14 | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cyclopentene-oxide + H2O | - |
Rhodococcus erythropolis | (R,R)-cyclopentene-1,2-diol + (S,S)-cyclopentene-1,2-diol | wild-type, 72% conversion, (R,R)-product with 14% enantiomeric excess | ? | |
cyclopentene-oxide + H2O | - |
Rhodococcus erythropolis DCL 14 | (R,R)-cyclopentene-1,2-diol + (S,S)-cyclopentene-1,2-diol | wild-type, 72% conversion, (R,R)-product with 14% enantiomeric excess | ? | |
rac-1-methyl-7-oxabicyclo[4.1.0]heptane + H2O | - |
Rhodococcus erythropolis | (1S,2S)-1-methylcyclohexane-1,2-diol + (1R,6S)-1-methyl-7-oxabicyclo[4.1.0]heptane | wild-type, 99% conversion, 19% enantiomeric excess for (1S,2S)-product | ? | |
rac-1-methyl-7-oxabicyclo[4.1.0]heptane + H2O | - |
Rhodococcus erythropolis DCL 14 | (1S,2S)-1-methylcyclohexane-1,2-diol + (1R,6S)-1-methyl-7-oxabicyclo[4.1.0]heptane | wild-type, 99% conversion, 19% enantiomeric excess for (1S,2S)-product | ? | |
rac-2-(phenoxymethyl)oxirane + H2O | - |
Rhodococcus erythropolis | (2S)-2-(phenoxymethyl)oxirane + (2R)-3-phenoxypropane-1,2-diol | wild-type, 33% conversion, 37% enantiomeric excess | ? | |
rac-2-(phenoxymethyl)oxirane + H2O | - |
Rhodococcus erythropolis DCL 14 | (2S)-2-(phenoxymethyl)oxirane + (2R)-3-phenoxypropane-1,2-diol | wild-type, 33% conversion, 37% enantiomeric excess | ? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
49 | - |
15 min, 50% resiudal activity for wild-type | Rhodococcus erythropolis |