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Literature summary for 3.3.2.8 extracted from

  • Zheng, H.; Reetz, M.T.
    Manipulating the stereoselectivity of limonene epoxide hydrolase by directed evolution based on iterative saturation mutagenesis (2010), J. Am. Chem. Soc., 132, 15744-15751.
    View publication on PubMed

Application

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

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

Organism UniProt Comment Textmining
Rhodococcus erythropolis
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Rhodococcus erythropolis DCL 14
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Substrates and Products (Substrate)

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
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Rhodococcus erythropolis (2S)-2-(phenoxymethyl)oxirane + (2R)-3-phenoxypropane-1,2-diol wild-type, 33% conversion, 37% enantiomeric excess ?
rac-2-(phenoxymethyl)oxirane + H2O
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Rhodococcus erythropolis DCL 14 (2S)-2-(phenoxymethyl)oxirane + (2R)-3-phenoxypropane-1,2-diol wild-type, 33% conversion, 37% enantiomeric excess ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
49
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15 min, 50% resiudal activity for wild-type Rhodococcus erythropolis