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Information on EC 3.3.2.8 - limonene-1,2-epoxide hydrolase
for references in articles please use BRENDA:EC3.3.2.8
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EC Tree 3 Hydrolases
3.3 Acting on ether bonds
3.3.2 Ether hydrolases
3.3.2.8 limonene-1,2-epoxide hydrolase
IUBMB Comments
Involved in the monoterpene degradation pathway of the actinomycete Rhodococcus erythropolis. The enzyme hydrolyses several alicyclic and 1-methyl-substituted epoxides, such as 1-methylcyclohexene oxide, indene oxide and cyclohexene oxide. It differs from the previously described epoxide hydrolases [EC 3.3.2.4 (trans-epoxysuccinate hydrolase), EC 3.3.2.6 (leukotriene-A4 hydrolase), EC 3.3.2.7 (hepoxilin-epoxide hydrolase), EC 3.3.2.9 (microsomal epoxide hydrolase) and EC 3.3.2.10 (soluble epoxide hydrolase)] as it is not inhibited by 2-bromo-4′-nitroacetophenone, diethyl dicarbonate, 4-fluorochalcone oxide or 1,10-phenanthroline. Both enantiomers of menth-8-ene-1,2-diol [i.e. (1R,2R,4S)-menth-8-ene-1,2-diol and (1S,2S,4R)-menth-8-ene-1,2-diol] are metabolized.
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
- 3.3.2.8
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stereoselective
- rhodococcus
- erythropolis
- hydrolases
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desymmetrization
- cyclohexene
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enantioselective
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alphabet
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regioselective
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lining
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six-stranded
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astronomically
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intricacies
- valpromide
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selenomethionine-substituted
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biocatalysis
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single-wavelength
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wide-ranging
- diols
- polyketide
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cyclopentene
- synthesis
showSynonyms
limonene-1,2-epoxide hydrolase, limonene epoxide hydrolase, ch55-leh, tomsk-leh, re-leh, limonene 1,2-epoxide hydrolase, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
LEH
limA
limonene 1,2-epoxide hydrolase
limonene epoxide hydrolase
limonene oxide hydrolase
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-
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limonene-1,2-epoxide hydrolase
Re-LEH
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
catalyzes simultaneous sequential and enantioconvergent epoxide conversion
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1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
one-step mechanism
1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
LEH mechanism, substrate specificity and stereoselectivity
1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
LEH mechanism, substrate specificity and stereoselectivity
1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
catalytic mechanism, quantum mechanics/molecular mechanics (QM/MM) free energy calculations for the reaction with molecular dynamics simulations
1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
catalytic mechanism analyzed by quantum mechanics/molecular mechanics (QM/MM) calculations, computational model, overview. Enzyme LEH reacts by a single-step concerted general acid-catalyzed mechanism, which is distinct from the two-step general base-catalyzed mechanism typical for the alpha/beta-hydrolase class of EHs. Overall, this mechanism is very similar to a borderline-SN2-type mechanism leading to nucleophilic attack at the more substituted oxirane carbon atom. Thus, no enzyme-substrate intermediate is detected during the experiments
1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
catalytic mechanism, quantum mechanics/molecular mechanics (QM/MM) free energy calculations for the reaction with molecular dynamics simulations
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1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
catalytic mechanism analyzed by quantum mechanics/molecular mechanics (QM/MM) calculations, computational model, overview. Enzyme LEH reacts by a single-step concerted general acid-catalyzed mechanism, which is distinct from the two-step general base-catalyzed mechanism typical for the alpha/beta-hydrolase class of EHs. Overall, this mechanism is very similar to a borderline-SN2-type mechanism leading to nucleophilic attack at the more substituted oxirane carbon atom. Thus, no enzyme-substrate intermediate is detected during the experiments
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1,2-epoxymenth-8-ene + H2O = menth-8-ene-1,2-diol
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of epoxide
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PATHWAY SOURCE
PATHWAYS
MetaCyc
limonene degradation I (D-limonene), limonene degradation II (L-limonene)
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SYSTEMATIC NAME
IUBMB Comments
1,2-epoxymeth-8-ene hydrolase
Involved in the monoterpene degradation pathway of the actinomycete Rhodococcus erythropolis. The enzyme hydrolyses several alicyclic and 1-methyl-substituted epoxides, such as 1-methylcyclohexene oxide, indene oxide and cyclohexene oxide. It differs from the previously described epoxide hydrolases [EC 3.3.2.4 (trans-epoxysuccinate hydrolase), EC 3.3.2.6 (leukotriene-A4 hydrolase), EC 3.3.2.7 (hepoxilin-epoxide hydrolase), EC 3.3.2.9 (microsomal epoxide hydrolase) and EC 3.3.2.10 (soluble epoxide hydrolase)] as it is not inhibited by 2-bromo-4'-nitroacetophenone, diethyl dicarbonate, 4-fluorochalcone oxide or 1,10-phenanthroline. Both enantiomers of menth-8-ene-1,2-diol [i.e. (1R,2R,4S)-menth-8-ene-1,2-diol and (1S,2S,4R)-menth-8-ene-1,2-diol] are metabolized.
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CAS REGISTRY NUMBER
COMMENTARY
216503-88-7
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(-)-limonene oxide + H2O
(1R,2R,4S)-limonene-1,2-diol
Substrates: -
Products: -
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?
(1R,2S)-1-methylcyclohexane oxide + H2O
(1S,2S)-1-methylcyclohexane-1,2-diol
Substrates: -
Products: -
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ir
(1S,2R)-1-methylcyclohexane oxide + H2O
(1R,2R)-1-methylcyclohexane-1,2-diol
Substrates: -
Products: -
Products: -
ir
(1S2R4S)-limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
-
Substrates: -
Products: optically pure, diastereomeric excess above 99%
Products: optically pure, diastereomeric excess above 99%
?
(4R)-limonene-1,2-epoxide + H2O
?
Substrates: the mixture of cis (1R,2S,4R) and trans (1S,2R,4R) isomers of (+)-limonene-1,2-epoxide and the mixture of cis (1S,2R,4S) and trans (1R,2S,4S) isomers of (-)-limonene-1,2-epoxide are quantitatively converted into the diaxial (1S,2S,4R)- and (1R,2R,4S)-limonene-1,2-diols, respectively. Cyclopentene-1,2-epoxide is no substrate for enzyme CH55-LEH. Enzyme substrate specificity and stereospecificity, overview
Products: -
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?
(4S)-limonene-1,2-epoxide + H2O
?
Substrates: the mixture of cis (1R,2S,4R) and trans (1S,2R,4R) isomers of (+)-limonene-1,2-epoxide and the mixture of cis (1S,2R,4S) and trans (1R,2S,4S) isomers of (-)-limonene-1,2-epoxide are quantitatively converted into the diaxial (1S,2S,4R)- and (1R,2R,4S)-limonene-1,2-diols, respectively. Cyclopentene-1,2-epoxide is no substrate for enzyme CH55-LEH. Enzyme substrate specificity and stereospecificity, overview
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?
1,2-epoxy-2,6-dimethyl-5-heptene + H2O
(2S)-2,6-dimethyl-5-hepten-1,2-diol
-
Substrates: -
Products: -
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?
1,2-epoxy-2-methyl-6-heptene + H2O
(2S)-2-methyl-6-heptene-1,2-diol
-
Substrates: -
Products: -
Products: -
?
1,2-epoxy-2-methylheptane + H2O
(2S)-2-methylheptane-1,2-diol
-
Substrates: -
Products: -
Products: -
?
1,2-epoxy-3-benzyl-2-methylpropane + H2O
(2S)-3-benzyl-2-methylpropane-1,2-diol
-
Substrates: -
Products: -
Products: -
?
1-methylcyclohexene oxide + H2O
(1S,2S)-1-methylcyclohexane-1,2-diol
-
Substrates: -
Products: -
Products: -
?
4-(1-methylethenyl)-cyclohexan-1,2-epoxide + H2O
4-(1-methylethenyl)-cyclohexan-1,2-diol
Substrates: -
Products: -
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?
4-(oxiran-2-yl)butan-1-ol + H2O
(oxan-2-yl)methanol
Substrates: -
Products: -
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?
cis-2,3-butene oxide + H2O
butane-2,3-diol
Substrates: -
Products: -
Products: -
?
cis-stilbene oxide + H2O
1,2-diphenylethane-1,2-diol
Substrates: -
Products: -
Products: -
?
cyclohexene oxide + H2O
(1S,2S)-trans cyclohexanediol + (1R,2R)-trans-cyclohexanediol
-
Substrates: -
Products: -
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?
cyclohexene-1,2-epoxide + H2O
(R1,R2)-cyclohexane-1,2-diol
-
Substrates: -
Products: -
Products: -
?
cyclohexene-1,2-epoxide + H2O
(S1,S2)-cyclohexane-1,2-diol
-
Substrates: -
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?
cyclohexenoxide + H2O
cyclohexan-1,2-diol
Substrates: -
Products: -
Products: -
?
cyclopentene oxide + H2O
cyclopentane-1,2-diol
Substrates: -
Products: -
Products: -
?
cyclopentene-1,2-epoxide + H2O
(R1,R2)-cyclopentane-1,2-diol
-
Substrates: -
Products: -
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?
cyclopentene-1,2-epoxide + H2O
(S1,S2)-cyclopentane-1,2-diol
-
Substrates: -
Products: -
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?
limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
Substrates: enantiomeric mixtures of (+)-limone oxide, overview. Enzyme CH55-LEH prefers the cis form of (-)-limonene oxide
Products: -
Products: -
?
limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol
Substrates: enantiomeric mixtures of (+)-limone oxide, overview. Enzyme Tomsk-LEH shows stereospecificity in the hydrolysis of cis/trans mixtures of (+)-limonene oxide by preferring the trans isomer
Products: -
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?
rac-1-methyl-7-oxabicyclo[4.1.0]heptane + H2O
(1S,2S)-1-methylcyclohexane-1,2-diol + (1R,6S)-1-methyl-7-oxabicyclo[4.1.0]heptane
rac-2-(phenoxymethyl)oxirane + H2O
(2S)-2-(phenoxymethyl)oxirane + (2R)-3-phenoxypropane-1,2-diol
styrene-7,8-oxide + H2O
styrene-7,8-diol
Substrates: -
Products: -
Products: -
?
(1R,2S,4R)-limonene-1,2-epoxide + H2O
(1R,2S,4R)-limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
(1R,2S,4R)-limonene-1,2-epoxide + H2O
(1R,2S,4R)-limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
(1R,2S,4R)-limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol
-
Substrates: -
Products: optically pure, diastereomeric excess above 99%
Products: optically pure, diastereomeric excess above 99%
?
(1R,2S,4R)-limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol
Substrates: the reaction mechanism involves epoxide protonation by Asp109, nucleophilic attack by water, and abstraction of a proton from water by Asp132. The isopropenyl group plays a crucial role because it restricts the half-chair conformation to one of the two possible helicities. In this conformation, attack on the different epoxide carbons will lead to either a chair-like or a twist-boat transition state structure, the latter resulting in a higher barrier. The regioselectivity is thus governed by conformational and not electronic factors
Products: -
Products: -
ir
(1R,2S,4R)-limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol
Substrates: the reaction mechanism involves epoxide protonation by Asp109, nucleophilic attack by water, and abstraction of a proton from water by Asp132. The isopropenyl group plays a crucial role because it restricts the half-chair conformation to one of the two possible helicities. In this conformation, attack on the different epoxide carbons will lead to either a chair-like or a twist-boat transition state structure, the latter resulting in a higher barrier. The regioselectivity is thus governed by conformational and not electronic factors
Products: -
Products: -
ir
(1R,2S,4S)-limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
-
Substrates: -
Products: optically pure, diastereomeric excess above 99%
Products: optically pure, diastereomeric excess above 99%
?
(1R,2S,4S)-limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
Substrates: the reaction mechanism involves epoxide protonation by Asp109, nucleophilic attack by water, and abstraction of a proton from water by Asp132. The isopropenyl group plays a crucial role because it restricts the half-chair conformation to one of the two possible helicities. In this conformation, attack on the different epoxide carbons will lead to either a chair-like or a twist-boat transition state structure, the latter resulting in a higher barrier. The regioselectivity is thus governed by conformational and not electronic factors
Products: -
Products: -
ir
(1R,2S,4S)-limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
Substrates: the reaction mechanism involves epoxide protonation by Asp109, nucleophilic attack by water, and abstraction of a proton from water by Asp132. The isopropenyl group plays a crucial role because it restricts the half-chair conformation to one of the two possible helicities. In this conformation, attack on the different epoxide carbons will lead to either a chair-like or a twist-boat transition state structure, the latter resulting in a higher barrier. The regioselectivity is thus governed by conformational and not electronic factors
Products: -
Products: -
ir
(1S,2R,4R)-limonene-1,2-epoxide + H2O
(1S,2R,4R)-limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
(1S,2R,4R)-limonene-1,2-epoxide + H2O
(1S,2R,4R)-limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
(1S,2R,4R)-limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol
-
Substrates: -
Products: optically pure, diastereomeric excess above 99%
Products: optically pure, diastereomeric excess above 99%
?
(1S,2R,4R)-limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol
Substrates: the reaction mechanism involves epoxide protonation by Asp109, nucleophilic attack by water, and abstraction of a proton from water by Asp132. The isopropenyl group plays a crucial role because it restricts the half-chair conformation to one of the two possible helicities. In this conformation, attack on the different epoxide carbons will lead to either a chair-like or a twist-boat transition state structure, the latter resulting in a higher barrier. The regioselectivity is thus governed by conformational and not electronic factors
Products: -
Products: -
ir
(1S,2R,4S)-limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
Substrates: the reaction mechanism involves epoxide protonation by Asp109, nucleophilic attack by water, and abstraction of a proton from water by Asp132. The isopropenyl group plays a crucial role because it restricts the half-chair conformation to one of the two possible helicities. In this conformation, attack on the different epoxide carbons will lead to either a chair-like or a twist-boat transition state structure, the latter resulting in a higher barrier. The regioselectivity is thus governed by conformational and not electronic factors
Products: -
Products: -
ir
(1S,2R,4S)-limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
Substrates: the reaction mechanism involves epoxide protonation by Asp109, nucleophilic attack by water, and abstraction of a proton from water by Asp132. The isopropenyl group plays a crucial role because it restricts the half-chair conformation to one of the two possible helicities. In this conformation, attack on the different epoxide carbons will lead to either a chair-like or a twist-boat transition state structure, the latter resulting in a higher barrier. The regioselectivity is thus governed by conformational and not electronic factors
Products: -
Products: -
ir
(4R)-limonene-1,2-epoxide + H2O
(4R)-limonene-1,2-diol
Substrates: -
Products: -
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?
(4R)-limonene-1,2-epoxide + H2O
(4R)-limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
(4S)-limonene-1,2-epoxide + H2O
(4S)-limonene-1,2-diol
Substrates: -
Products: -
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?
(4S)-limonene-1,2-epoxide + H2O
(4S)-limonene-1,2-diol
Substrates: -
Products: -
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?
1-methylcyclohexene oxide + H2O
1-methylcyclohexane-1,2-diol
Substrates: -
Products: -
Products: -
?
1-methylcyclohexene oxide + H2O
1-methylcyclohexane-1,2-diol
Substrates: -
Products: -
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?
2 cyclohexene-1,2-epoxide + 2 H2O
(S,S)-cyclohexane-1,2-diol + (R,R)-cyclohexane-1,2-diol
Substrates: LEH is the catalyst in the hydrolytic desymmetrization of cyclohexene oxide with formation of (R,R)- and (S,S)-cyclohexene-1,2-diol. Wild-type LEH shows an enanioselectivity of 2% enantiomeric excess (S,S), analysis of (R,R)- and (S,S)-selective LEH mutant variants (80-94% enantiomeric excess)
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2 cyclohexene-1,2-epoxide + 2 H2O
(S,S)-cyclohexane-1,2-diol + (R,R)-cyclohexane-1,2-diol
Substrates: LEH is the catalyst in the hydrolytic desymmetrization of cyclohexene oxide with formation of (R,R)- and (S,S)-cyclohexene-1,2-diol. Wild-type LEH shows an enanioselectivity of 2% enantiomeric excess (S,S), analysis of (R,R)- and (S,S)-selective LEH mutant variants (80-94% enantiomeric excess)
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2-butyloxirane + H2O
hexane-1,2-diol
Substrates: -
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2-butyloxirane + H2O
hexane-1,2-diol
Substrates: -
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cycloheptene-1,2-epoxide + H2O
cycloheptene-1,2-diol
Substrates: -
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cycloheptene-1,2-epoxide + H2O
cycloheptene-1,2-diol
Substrates: -
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cyclohexene oxide + H2O
cyclohexane-1,2-diol
Substrates: -
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cyclohexene oxide + H2O
cyclohexane-1,2-diol
Substrates: -
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cyclohexene-1,2-epoxide + H2O
cyclohexane-1,2-diol
Substrates: -
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cyclohexene-1,2-epoxide + H2O
cyclohexane-1,2-diol
Substrates: 0.25% activity compared to limonene-1,2-epoxide
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cyclohexene-1,2-epoxide + H2O
cyclohexane-1,2-diol
Substrates: 0.25% activity compared to limonene-1,2-epoxide
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?
cyclohexene-1,2-epoxide + H2O
cyclohexane-1,2-diol
Substrates: -
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cyclopentene-1,2-epoxide + H2O
cyclopentane-1,2-diol
Substrates: low activity
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cyclopentene-1,2-epoxide + H2O
cyclopentane-1,2-diol
Substrates: low activity
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cyclopentene-oxide + H2O
(R,R)-cyclopentene-1,2-diol + (S,S)-cyclopentene-1,2-diol
-
Substrates: -
Products: wild-type, 72% conversion, (R,R)-product with 14% enantiomeric excess
Products: wild-type, 72% conversion, (R,R)-product with 14% enantiomeric excess
?
cyclopentene-oxide + H2O
(R,R)-cyclopentene-1,2-diol + (S,S)-cyclopentene-1,2-diol
-
Substrates: -
Products: wild-type, 72% conversion, (R,R)-product with 14% enantiomeric excess
Products: wild-type, 72% conversion, (R,R)-product with 14% enantiomeric excess
?
indene oxide + H2O
indane-1,2-diol
Substrates: -
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indene oxide + H2O
indane-1,2-diol
Substrates: -
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limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol + (1R,2R,4S)-limonene-1,2-diol
Substrates: enantiomeric mixtures of (+)-limone oxide, overview. Re-LEH catalyzes the quantitative conversion of the (+)- and (-)-limonene oxide cis/trans mixtures into the respective diols, the (+)-enantiomer is the better substrate
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limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol + (1R,2R,4S)-limonene-1,2-diol
Substrates: enantiomeric mixtures of (+)-limone oxide, overview. Re-LEH catalyzes the quantitative conversion of the (+)- and (-)-limonene oxide cis/trans mixtures into the respective diols, the (+)-enantiomer is the better substrate
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limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: part of limonene degradation pathway which allows the organism to grow on limone as sole source of carbon and energy
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limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
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limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
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limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: limonene-1,2-epoxide is not the natural substrate of Tomsk-LEH
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limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: part of limonene degradation pathway which allows the organism to grow on limone as sole source of carbon and energy
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limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
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ir
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
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limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
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limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
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Products: -
?
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: limonene-1,2-epoxide is not the natural substrate of Tomsk-LEH
Products: -
Products: -
?
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: limonene-1,2-epoxide is not the natural substrate of CH55-LEH
Products: -
Products: -
?
phenylethylenoxide + H2O
1-phenylethane-1,2-diol
Substrates: -
Products: -
Products: -
?
phenylethylenoxide + H2O
1-phenylethane-1,2-diol
Substrates: -
Products: -
Products: -
?
rac-1-methyl-7-oxabicyclo[4.1.0]heptane + H2O
(1S,2S)-1-methylcyclohexane-1,2-diol + (1R,6S)-1-methyl-7-oxabicyclo[4.1.0]heptane
-
Substrates: -
Products: wild-type, 99% conversion, 19% enantiomeric excess for (1S,2S)-product
Products: wild-type, 99% conversion, 19% enantiomeric excess for (1S,2S)-product
?
rac-1-methyl-7-oxabicyclo[4.1.0]heptane + H2O
(1S,2S)-1-methylcyclohexane-1,2-diol + (1R,6S)-1-methyl-7-oxabicyclo[4.1.0]heptane
-
Substrates: -
Products: wild-type, 99% conversion, 19% enantiomeric excess for (1S,2S)-product
Products: wild-type, 99% conversion, 19% enantiomeric excess for (1S,2S)-product
?
rac-2-(phenoxymethyl)oxirane + H2O
(2S)-2-(phenoxymethyl)oxirane + (2R)-3-phenoxypropane-1,2-diol
-
Substrates: -
Products: wild-type, 33% conversion, 37% enantiomeric excess
Products: wild-type, 33% conversion, 37% enantiomeric excess
?
rac-2-(phenoxymethyl)oxirane + H2O
(2S)-2-(phenoxymethyl)oxirane + (2R)-3-phenoxypropane-1,2-diol
-
Substrates: -
Products: wild-type, 33% conversion, 37% enantiomeric excess
Products: wild-type, 33% conversion, 37% enantiomeric excess
?
additional information
?
-
Substrates: enzyme substrate specificity and stereospecificity, overview
Products: -
Products: -
?
additional information
?
-
-
Substrates: enzyme substrate specificity and stereospecificity, overview
Products: -
Products: -
?
additional information
?
-
Substrates: enantioselectivity and activity of limonene epoxide hydrolase, overview
Products: -
Products: -
?
additional information
?
-
Substrates: proposed hydrolysis mechanism, the Asp101-Arg99-Asp132 triad with a water molecule is regarded as the active central, overview
Products: -
Products: -
?
additional information
?
-
Substrates: commercially available cis/trans mixtures of (+)-limonene oxide (59:41 mixture of (1R,2S,4R) and (1S,2R,4R)) and (-)-limonene oxide (55:45 mixture of (1S,2R,4S) and (1R,2S,4S)) are dissolved in CH3CN and diluted with the appropriate LEH-containing buffer solution
Products: -
Products: -
?
additional information
?
-
Substrates: enantioselectivity and activity of limonene epoxide hydrolase, overview
Products: -
Products: -
?
additional information
?
-
Substrates: proposed hydrolysis mechanism, the Asp101-Arg99-Asp132 triad with a water molecule is regarded as the active central, overview
Products: -
Products: -
?
additional information
?
-
Substrates: commercially available cis/trans mixtures of (+)-limonene oxide (59:41 mixture of (1R,2S,4R) and (1S,2R,4R)) and (-)-limonene oxide (55:45 mixture of (1S,2R,4S) and (1R,2S,4S)) are dissolved in CH3CN and diluted with the appropriate LEH-containing buffer solution
Products: -
Products: -
?
additional information
?
-
Substrates: commercially available cis/trans mixtures of (+)-limonene oxide (59:41 mixture of (1R,2S,4R) and (1S,2R,4R)) and (-)-limonene oxide (55:45 mixture of (1S,2R,4S) and (1R,2S,4S)) are dissolved in CH3CN and diluted with the appropriate LEH-containing buffer solution
Products: -
Products: -
?
additional information
?
-
Substrates: commercially available cis/trans mixtures of (+)-limonene oxide (59:41 mixture of (1R,2S,4R) and (1S,2R,4R)) and (-)-limonene oxide (55:45 mixture of (1S,2R,4S) and (1R,2S,4S)) are dissolved in CH3CN and diluted with the appropriate LEH-containing buffer solution
Products: -
Products: -
?
additional information
?
-
Substrates: the mixture of cis (1R,2S,4R) and trans (1S,2R,4R) isomers of (+)-limonene-1,2-epoxide and the mixture of cis (1S,2R,4S) and trans (1R,2S,4S) isomers of (-)-limonene-1,2-epoxide are quantitatively converted into the diaxial (1S,2S,4R)- and (1R,2R,4S)-limonene-1,2-diols, respectively. Enzyme substrate specificity and stereospecificity, overview
Products: -
Products: -
?
additional information
?
-
Substrates: the mixture of cis (1R,2S,4R) and trans (1S,2R,4R) isomers of (+)-limonene-1,2-epoxide and the mixture of cis (1S,2R,4S) and trans (1R,2S,4S) isomers of (-)-limonene-1,2-epoxide are quantitatively converted into the diaxial (1S,2S,4R)- and (1R,2R,4S)-limonene-1,2-diols, respectively. Enzyme substrate specificity and stereospecificity, overview
Products: -
Products: -
?
additional information
?
-
-
Substrates: stereochemistry and catalytic mechanism, overview
Products: -
Products: -
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(1R,2S,4R)-limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol
-
Substrates: -
Products: optically pure, diastereomeric excess above 99%
Products: optically pure, diastereomeric excess above 99%
?
(1R,2S,4S)-limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
-
Substrates: -
Products: optically pure, diastereomeric excess above 99%
Products: optically pure, diastereomeric excess above 99%
?
(1S,2R,4R)-limonene-1,2-epoxide + H2O
(1S,2S,4R)-limonene-1,2-diol
-
Substrates: -
Products: optically pure, diastereomeric excess above 99%
Products: optically pure, diastereomeric excess above 99%
?
(1S2R4S)-limonene-1,2-epoxide + H2O
(1R,2R,4S)-limonene-1,2-diol
-
Substrates: -
Products: optically pure, diastereomeric excess above 99%
Products: optically pure, diastereomeric excess above 99%
?
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: part of limonene degradation pathway which allows the organism to grow on limone as sole source of carbon and energy
Products: -
Products: -
ir
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: part of limonene degradation pathway which allows the organism to grow on limone as sole source of carbon and energy
Products: -
Products: -
ir
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
limonene-1,2-epoxide + H2O
limonene-1,2-diol
Substrates: -
Products: -
Products: -
?
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
poly(ethylene glycol)
enzyme binding structure, overview
valpromide
binds at the active site, enzyme binding structure, overview
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3
cis-2,3-butene oxide
mutant M32A/M78I/I80F/L103I/I116V/F139L, pH 8, 30°C
-
3 - 5
cis-2,3-butene oxide
mutant L74I/L103V/F134Y/F139W, pH 8, 30°C
-
344
cis-2,3-butene oxide
mutant M32L/L103V/L114A/I116F/F139W, pH 8, 30°C
-
0.06
cis-stilbene oxide
mutant M32L/L35G/I80W/L103V/F139L, pH 8, 30°C
0.16
cis-stilbene oxide
mutant M32L/L35M/M78I/L103I/L114M/I116F, pH 8, 30°C
0.19
cis-stilbene oxide
mutant M32L/L35M/L103I/L114M/I116F/F139L, pH 8, 30°C
6.39
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K/E45D/I80V/E124D
6.7
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant wild-type enzyme
6.96
cyclohexene-1,2-epoxide
pH 7.4, 37°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K/E45D/I80V/E124D
8.23
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K
13.09
cyclohexene-1,2-epoxide
pH 7.4, 37°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F/E45D
14.41
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V
15.44
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F
16.06
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V
16.1
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F/E45D
16.97
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F
19.41
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F
225
cyclopentene oxide
mutant L35W/L74F/I80G/I116V/F139L, pH 8, 30°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.005
cis-2,3-butene oxide
mutant M32A/M78I/I80F/L103I/I116V/F139L, pH 8, 30°C
-
0.017
cis-2,3-butene oxide
mutant L74I/L103V/F134Y/F139W, pH 8, 30°C
-
0.022
cis-2,3-butene oxide
mutant M32L/L103V/L114A/I116F/F139W, pH 8, 30°C
-
0.002
cis-stilbene oxide
mutant M32L/L35M/L103I/L114M/I116F/F139L, pH 8, 30°C
0.003
cis-stilbene oxide
mutant M32L/L35M/M78I/L103I/L114M/I116F, pH 8, 30°C
0.052
cis-stilbene oxide
mutant M32L/L35G/I80W/L103V/F139L, pH 8, 30°C
0.59
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K
0.64
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K/E45D/I80V/E124D
0.76
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F
0.82
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant wild-type enzyme
0.82
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F
0.84
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F/E45D
0.84
cyclohexene-1,2-epoxide
pH 7.4, 37°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K/E45D/I80V/E124D
0.86
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V
0.93
cyclohexene-1,2-epoxide
pH 7.4, 37°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F/E45D
1.25
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F
1.55
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V
0.063
cyclopentene oxide
mutant L35W/L74F/I80G/I116V/F139L, pH 8, 30°C
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00006
cis-2,3-butene oxide
mutant M32L/L103V/L114A/I116F/F139W, pH 8, 30°C
-
0.00048
cis-2,3-butene oxide
mutant L74I/L103V/F134Y/F139W, pH 8, 30°C
-
0.0019
cis-2,3-butene oxide
mutant M32A/M78I/I80F/L103I/I116V/F139L, pH 8, 30°C
-
0.0105
cis-stilbene oxide
mutant M32L/L35M/L103I/L114M/I116F/F139L, pH 8, 30°C
0.019
cis-stilbene oxide
mutant M32L/L35M/M78I/L103I/L114M/I116F, pH 8, 30°C
0.89
cis-stilbene oxide
mutant M32L/L35G/I80W/L103V/F139L, pH 8, 30°C
0.042
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F
0.049
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F
0.052
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F/E45D
0.054
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V
0.071
cyclohexene-1,2-epoxide
pH 7.4, 37°C, recombinant mutant T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F/E45D
0.072
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K
0.074
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F
0.1
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K/E45D/I80V/E124D
0.108
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant mutant T76K/L114V/I116V
0.121
cyclohexene-1,2-epoxide
pH 7.4, 37°C, recombinant mutant S15P/M78F/N92K/F139V/T76K/T85K/E45D/I80V/E124D
0.122
cyclohexene-1,2-epoxide
pH 7.4, 30°C, recombinant wild-type enzyme
0.00028
cyclopentene oxide
mutant L35W/L74F/I80G/I116V/F139L, pH 8, 30°C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.097
Tomsk-LEH, substrate (+)-limonene oxide, pH 8.0, 20°C, non-optimized conditions
0.157
Tomsk-LEH, substrate (+)-limonene oxide, pH 8.0, 30°C, optimized conditions
0.23
CH55-LEH , substrate (-)-limonene oxide, pH 8.0, 20°C, non-optimized conditions
1.33
CH55-LEH , substrate (-)-limonene oxide, pH 8.0, 50°C, optimized conditions
1.85
Re-LEH , substrate (-)-limonene oxide, pH 8.0, 20°C, non-optimized conditions
13.95
Re-LEH , substrate (+)-limonene oxide, pH 8.0, 50°C, optimized conditions
3.17
Re-LEH , substrate (-)-limonene oxide, pH 8.0, 50°C, optimized conditions
6.18
Re-LEH , substrate (+)-limonene oxide, pH 8.0, 20°C, non-optimized conditions
85.1
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 10.5
-
below pH 5 chemical hydrolysis overtook biological hydrolysis rate
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
50
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
physiological function
additional information
malfunction
replacement of the catalytic aspartic acid residue to alanine (D80A) completely abolishes activity towards the tested substrates cyclohexene epoxide and (+)-limonene epoxide
malfunction
replacement of th catalytic aspartic acid residue to alanine (D82A) completely abolishes activity towards the tested substrates cyclohexene epoxide and (+)-limonene epoxide
physiological function
limonene-1,2-epoxide hydrolases (LEHs), a subset of the epoxide hydrolase family, present interesting opportunities for the mild, regio- and stereo- selective hydrolysis of epoxide substrates. LEHs show moderate enantioselectivity for non-natural ligands, combined with narrow substrate specificity
physiological function
epoxide hydrolases (EHs) catalyze the hydrolysis of epoxides to vicinal diols. EHs are found in all types of living organisms, including mammals, invertebrates, plants, bacteria and fungi. They have three main functions: detoxification, metabolism, and synthesis of signaling molecules
physiological function
-
limonene-1,2-epoxide hydrolases (LEHs), a subset of the epoxide hydrolase family, present interesting opportunities for the mild, regio- and stereo- selective hydrolysis of epoxide substrates. LEHs show moderate enantioselectivity for non-natural ligands, combined with narrow substrate specificity
-
physiological function
-
epoxide hydrolases (EHs) catalyze the hydrolysis of epoxides to vicinal diols. EHs are found in all types of living organisms, including mammals, invertebrates, plants, bacteria and fungi. They have three main functions: detoxification, metabolism, and synthesis of signaling molecules
-
additional information
the enzyme's catalytic mechanism is different from that of the epoxide hydrolases, EHs, belonging to the alpha/beta-hydrolase superfamily. The LEH enzyme active site contains three residues (Asp101, Arg99, and Asp132) that act in a concerted fashion to activate a water molecule which is able to open the epoxide ring without the formation of a covalently bound alkyl-enzyme intermediate. Importance of the catalytic Asp80 residues for the enzymatic activity of Tomsk-LEH. The LEH substrate binding pocket appears to have high affinity for polar molecules and additional electron density is observed in the active site pocket in the different LEH structure. Active site structure, overview
additional information
the enzyme's catalytic mechanism is different from that of the epoxide hydrolases, EHs, belonging to the alpha/beta-hydrolase superfamily. The LEH enzyme active site contains three residues (Asp101, Arg99, and Asp132) that act in a concerted fashion to activate a water molecule which is able to open the epoxide ring without the formation of a covalently bound alkyl-enzyme intermediate. Importance of the catalytic Asp80 residues for the enzymatic activity of Tomsk-LEH. The LEH substrate binding pocket appears to have high affinity for polar molecules and additional electron density is observed in the active site pocket in the different LEH structure. Active site structure, overview
additional information
the enzyme's catalytic mechanism is different from that of the epoxide hydrolases, EHs, belonging to the alpha/beta-hydrolase superfamily. The LEH enzyme active site contains three residues (Asp101, Arg99, and Asp132) that act in a concerted fashion to activate a water molecule which is able to open the epoxide ring without the formation of a covalently bound alkyl-enzyme intermediate. Importance of the catalytic Asp82 residues for the enzymatic activity of CH55-LEH. The LEH substrate binding pocket appears to have high affinity for polar molecules and additional electron density is observed in the active site pocket in the different LEH structure. Active site structure, overview
additional information
the enzyme's catalytic mechanism is different from that of the epoxide hydrolases, EHs, belonging to the alpha/beta-hydrolase superfamily. The LEH enzyme active site contains three residues (Asp101, Arg99, and Asp132) that act in a concerted fashion to activate a water molecule which is able to open the epoxide ring without the formation of a covalently bound alkyl-enzyme intermediate. Importance of the catalytic Asp82 residues for the enzymatic activity of CH55-LEH. The LEH substrate binding pocket appears to have high affinity for polar molecules and additional electron density is observed in the active site pocket in the different LEH structure. Active site structure, overview
additional information
the N-terminal extension of Re-LEH involved in the intersubunit interface which increases the buried surface area. The LEH monomer fold contains a curved six-stranded mixed beta-sheet, with three alpha-helices packed onto its concave side to form the active site pocket. Active site structure, overview
additional information
-
the N-terminal extension of Re-LEH involved in the intersubunit interface which increases the buried surface area. The LEH monomer fold contains a curved six-stranded mixed beta-sheet, with three alpha-helices packed onto its concave side to form the active site pocket. Active site structure, overview
additional information
-
structural and computational insight into the catalytic mechanism of limonene epoxide hydrolase variants in stereoselective transformations, molecular dynamics simulations
additional information
quantum mechanics/molecular mechanics (QM/MM) free energy calculations for the reaction with molecular dynamics simulations of the enzyme internal dynamics, and the calculation of binding affinities (using the WaterSwap method) for various representatives of the enzyme conformational ensemble, show that the presence of natural or non-natural substrates differentially modulates the dynamic and catalytic behavior of LEH. The cross-talk between the protein and the ligands favors the selection of specific substrate-dependent interactions in the binding site, priming reactive complexes to select different preferential reaction pathways. LEH substrate binding pocket structure, LEH forms a stable homodimer, and each monomer can bind a substrate molecule within its catalytic pocket, overview. Crucial role of monomer-monomer interactions in stabilizing and tuning LEH dynamics and stability. Hydrolysis by LEH occurs via a complex mechanism, the Asp101-Arg99-Asp132 triad drives a concerted reaction involving the deprotonation of a water molecule by Asp132, the nucleophilic attack of the resulting hydroxide ion on the epoxide and protonation of the oxirane ring by the protonated Asp101 (specifically labeled Ash101). Arg99 is strongly associated through hydrogen bonds and electrostatic interactions with both Asp101 and Asp132 and even if it is not directly involved in the reaction mechanism, its mutation results in a deactivated enzyme. This complex picture is completed by the proper positioning and activation of the nucleophilic water by the H-bond network formed by Asn55 and Tyr53 (and Asp132 itself). In the model, the side chains of the residues of the catalytic triad, the water molecule and the epoxide are included in the QM region. The opening of the epoxide can result from the attack on either of the two carbon atoms of the LEO oxirane ring. Experimental evidence indicates that the water molecule privileges the attack on the more substituted C1 atom
additional information
reaction quantum mechanics/molecular mechanics (QM/MM) calculations, molecular dynamics simulations and active site structures of wild-type and mutant enzymes, overview. Rhodococcus erythropolis DCL14 LEH has the exceptionally low molecular mass of 16 kDa, which is too small to contain any of the highly conserved motifs of the catalytic triad used by alpha/beta-hydrolase folded EHs. LEH has a narrow substrate range compared to other EHs
additional information
-
quantum mechanics/molecular mechanics (QM/MM) free energy calculations for the reaction with molecular dynamics simulations of the enzyme internal dynamics, and the calculation of binding affinities (using the WaterSwap method) for various representatives of the enzyme conformational ensemble, show that the presence of natural or non-natural substrates differentially modulates the dynamic and catalytic behavior of LEH. The cross-talk between the protein and the ligands favors the selection of specific substrate-dependent interactions in the binding site, priming reactive complexes to select different preferential reaction pathways. LEH substrate binding pocket structure, LEH forms a stable homodimer, and each monomer can bind a substrate molecule within its catalytic pocket, overview. Crucial role of monomer-monomer interactions in stabilizing and tuning LEH dynamics and stability. Hydrolysis by LEH occurs via a complex mechanism, the Asp101-Arg99-Asp132 triad drives a concerted reaction involving the deprotonation of a water molecule by Asp132, the nucleophilic attack of the resulting hydroxide ion on the epoxide and protonation of the oxirane ring by the protonated Asp101 (specifically labeled Ash101). Arg99 is strongly associated through hydrogen bonds and electrostatic interactions with both Asp101 and Asp132 and even if it is not directly involved in the reaction mechanism, its mutation results in a deactivated enzyme. This complex picture is completed by the proper positioning and activation of the nucleophilic water by the H-bond network formed by Asn55 and Tyr53 (and Asp132 itself). In the model, the side chains of the residues of the catalytic triad, the water molecule and the epoxide are included in the QM region. The opening of the epoxide can result from the attack on either of the two carbon atoms of the LEO oxirane ring. Experimental evidence indicates that the water molecule privileges the attack on the more substituted C1 atom
-
additional information
-
reaction quantum mechanics/molecular mechanics (QM/MM) calculations, molecular dynamics simulations and active site structures of wild-type and mutant enzymes, overview. Rhodococcus erythropolis DCL14 LEH has the exceptionally low molecular mass of 16 kDa, which is too small to contain any of the highly conserved motifs of the catalytic triad used by alpha/beta-hydrolase folded EHs. LEH has a narrow substrate range compared to other EHs
-
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). A0A0G3ICV8_9ZZZZ
125
0
14085
TrEMBL
other Location (Reliability: 1)
EPHG_MYCTU
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
149
0
16593
Swiss-Prot
Secretory Pathway (Reliability: 3)
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
dimer
the LEH monomer fold contains a curved six-stranded mixed beta-sheet, with three alpha-helices packed onto its concave side to form the active site pocket
dimer
LEH forms a stable homodimer, and each monomer can bind a substrate molecule within its catalytic pocket
dimer
-
LEH forms a stable homodimer, and each monomer can bind a substrate molecule within its catalytic pocket
-
dimer
the LEH monomer fold contains a curved six-stranded mixed beta-sheet, with three alpha-helices packed onto its concave side to form the active site pocket
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified enzyme mutant H-2-H5 (E45D/L74F/T76K/M78F/N92K/L114V/I116V) and mutant H-2-H5 in complex with (S,S)-cyclohexenediol, sitting-drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 50 mM potassium phosphate buffer, pH 8.0, with 0.001 ml of reservoir solution containing 0.1 M HEPES, pH 7.5, 2% v/v PEG 400, and 2.0 M ammonium sulfate, and equilibration against reservoir solution, 20°C, the single protein crystals are soaked in 100 mM ligand solution for 1 min, X-ray diffraction structure determination and analysis, molecular replacement method using the wild-type LEH (PDB ID 1NU3) as a search model, comparison of crystal structures of wild-type and mutant enzymes
mutant SZ348, sitting-drop vapor diffusion method, crystallized in 2.4 M sodium/potassium phosphate, 0.1 M Tris-HCl, pH 8.5, mixing of 0.002 ml of 16 mg/ml protein solution with 0.002 ml of reservoir solution, equilibration against the reservoir solution. The complex crystals of SZ348-CYO1 are made by soaking the crystals of SZ348 in sodium/potassium phosphate, 0.1 M Tris-HCl, pH 8.5, 10 mM of cyclopentene-1,2-epoxide, and 2.5% v/v acetonitrile for 5-10 min, at 18°C, X-ray diffraction structure determination and analysis
-
purified recombinant enzyme in apoform or in complex with inhibitor poly(ethylene glycol), 10 mg/ml protein solution is mixed with an equal volume of precipitant solution and is covered with a 1:1 mix of silicon and paraffin oils, X-ray diffraction structure determination and analysis at 1.42-1.47 A resolution
purified recombinant enzyme in apoform or in complex with inhibitor valpromide, 10 mg/ml protein solution is mixed with an equal volume of precipitant solution and is covered with a 1:1 mix of silicon and paraffin oils, X-ray diffraction structure determination and analysis at 1.16-1.26 A resolution
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D101A
catalytically inactive, D101 is the acid catalyst that protonates the epoxide oxygen
D101N
catalytically inactive, D101 is the acid catalyst that protonates the epoxide oxygen
D132N
mutant shows complete suppression of the hydrolytic activity
E45D/L74F/T76K/M78F/N92K/L114V/I116V
site-directed mutagenesis, the mutant lacking the N- and C-terminal mutations displays 86% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol
I5C/S15P/A19K/T76K/E84C/T85V/G89C/S91C/N92K/Y96F/E124D
site-directed mutagenesis, the multisite mutant shows enhanced and inverted enantioselectivity, and an increase in apparent melting temperature relative to wild-type LEH from 50 to 85°C and a more than 250fold longer half-life
L114C/I116V
-
substrate cyclopentene-oxide, 72% conversion, (S,S)-product with 68% enantiomeric excess
L114I/I116V
-
substrate cyclopentene-oxide, 74% conversion, (S,S)-product with 50% enantiomeric excess
L114V/I116V
-
substrate cyclopentene-oxide, 72% conversion, (S,S)-product with 60% enantiomeric excess
L35W/L74F/I80G/I116V/F139L
mutant synthesizes (1S,2S)-cyclopentane-1,2-diol with 82% ee
L74I/I80C
-
substrate cyclopentene-oxide, 75% conversion, (R,R)-product with 66% enantiomeric excess
L74I/I80V
-
substrate cyclopentene-oxide, 75% conversion, (R,R)-product with 58% enantiomeric excess
L74I/L103V/F134Y/F139W
mutant synthesizes (1R,2R)-butane-2,3-diol with 8% ee
L74V/I80V
-
substrate cyclopentene-oxide, 67% conversion, (R,R)-product with 53% enantiomeric excess
M32A/M78I/I80F/L103I/I116V/F139L
mutant synthesizes (1S,2S)-tane-2,3-diol with 85% ee
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
M32L/L103V/L114A/I116F/F139W
mutant synthesizes (1R,2R)-butane-2,3-diol with 34% ee
M32L/L35C
-
substrate cyclopentene-oxide, 78% conversion, (S,S)-product with 16% enantiomeric excess
M32L/L35F
-
substrate cyclopentene-oxide, 79% conversion, (S,S)-product with 24% enantiomeric excess
M32L/L35G/I80W/L103V/F139L
mutant synthesizes (1R,2R)-diphenylethane-1,2-diol with >99% ee
M32L/L35M/L103I/L114M/I116F/F139L
mutant synthesizes (1S,2S)-diphenylethane-1,2-diol with 94% ee
M32L/L35M/M78I/L103I/L114M/I116F
mutant synthesizes (1S,2S)-diphenylethane-1,2-diol with 89% ee
M32L/L35V
-
substrate cyclopentene-oxide, 78% conversion, (S,S)-product with 10% enantiomeric excess
M78F/V83I
-
substrate cyclopentene-oxide, 82% conversion, (R,R)-product with 29% enantiomeric excess
M78I/V83I
-
substrate cyclopentene-oxide, 80% conversion, (R,R)-product with 13% enantiomeric excess
M78V/V83I
-
substrate cyclopentene-oxide, 68% conversion, (R,R)-product with 7% enantiomeric excess
N55A
R99K
S15P/A19K/E45K/T76K/T85V/N92K/Y96F/E124D
site-directed mutagenesis, the multisite mutant shows enhanced and inverted enantioselectivity, and an increase in apparent melting temperature relative to wild-type LEH from 50 to 85°C and a more than 250fold longer half-life, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 46°C and the enantiomeric excess is 80% in favor of (R,R)-cyclohexene-1,2-diol
S15P/M78F
site-directed mutagenesis, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 47°C and the enantiomeric excess is 34% in favor of (R,R)-cyclohexene-1,2-diol
S15P/M78F/N92K/F139V
site-directed mutagenesis, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 48°C and the enantiomeric excess is 39% in favor of (R,R)-cyclohexene-1,2-diol
S15P/M78F/N92K/F139V/T76K/T85K
site-directed mutagenesis, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 44°C and the enantiomeric excess is 45% in favor of (R,R)-cyclohexene-1,2-diol
S15P/M78F/N92K/F139V/T76K/T85K/E45D/I80V/E124D
site-directed mutagenesis, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 46°C and the enantiomeric excess is 80% in favor of (R,R)-cyclohexene-1,2-diol
T76D/L114V/I116V
site-directed mutagenesis, the mutant lacking the N- and C-terminal mutations, maintains enantioselectivity of 71% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol
T76K/L114V/I116V
site-directed mutagenesis, the mutant lacking the N- and C-terminal mutations, maintains enantioselectivity of 71% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 44°C
T76K/L114V/I116V/F139V/L147F
site-directed mutagenesis, the mutant lacking the N-terminal mutations shows enantioselectivities of 82% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 45°C
T76K/L114V/I116V/N92D/F139V/L147F
site-directed mutagenesis, the mutant lacking the N-terminal mutations shows enantioselectivities of 82% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol
T76K/L114V/I116V/N92K/F139V/L147F
site-directed mutagenesis, the mutant lacking the N-terminal mutations shows enantioselectivities of 83% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol
T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 51°C, and the enantiomeric excess is 92% in favor of (S,S)-cyclohexene-1,2-diol
T76K/L114V/I116V/N92K/F139V/L147F/S15D/A19K/L74F/M78F/E45D
site-directed mutagenesis, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 51°C and the enantiomeric excess is 94% in favor of (S,S)-cyclohexene-1,2-diol
Y53F
Y53F/N55A
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 78% (oxan-2-yl)methanol plus 10% hexane-1,2,6-triol
Y53F/N55A/I116V
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 87% (oxan-2-yl)methanol plus 13% hexane-1,2,6-triol
Y53F/N55A/I80F/I116V
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 54% (oxan-2-yl)methanol plus 1% hexane-1,2,6-triol
Y53F/N55A/I80F/L114V/I116V
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 60% (oxan-2-yl)methanol plus 1% hexane-1,2,6-triol
Y53F/N55P
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 71% (oxan-2-yl)methanol plus 10% hexane-1,2,6-triol
Y53I/N55A
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 65% (oxan-2-yl)methanol plus 4% hexane-1,2,6-triol
Y53I/N55P
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 70% (oxan-2-yl)methanol plus 6% hexane-1,2,6-triol
Y53L/N55P
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 54% (oxan-2-yl)methanol plus 4% hexane-1,2,6-triol
Y53V/N55P
with substrate 4-(oxiran-2-yl)butan-1-ol, mutant generates about 28% (oxan-2-yl)methanol plus 3% hexane-1,2,6-triol
L74V/I80V
-
substrate cyclopentene-oxide, 67% conversion, (R,R)-product with 53% enantiomeric excess
-
M32L/L35C
-
substrate cyclopentene-oxide, 78% conversion, (S,S)-product with 16% enantiomeric excess
-
M32L/L35F
-
substrate cyclopentene-oxide, 79% conversion, (S,S)-product with 24% enantiomeric excess
-
M32L/L35V
-
substrate cyclopentene-oxide, 78% conversion, (S,S)-product with 10% enantiomeric excess
-
E45D/L74F/T76K/M78F/N92K/L114V/I116V
-
site-directed mutagenesis, the mutant lacking the N- and C-terminal mutations displays 86% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol
-
I5C/S15P/A19K/T76K/E84C/T85V/G89C/S91C/N92K/Y96F/E124D
-
site-directed mutagenesis, the multisite mutant shows enhanced and inverted enantioselectivity, and an increase in apparent melting temperature relative to wild-type LEH from 50 to 85°C and a more than 250fold longer half-life
-
N55A
-
site-directed mutagenesis, QM/MM-optimized active structure of the enzyme mutant compared to wild-type
-
R99K
-
site-directed mutagenesis, QM/MM-optimized active structure of the enzyme mutant compared to wild-type
-
S15P/A19K/E45K/T76K/T85V/N92K/Y96F/E124D
-
site-directed mutagenesis, the multisite mutant shows enhanced and inverted enantioselectivity, and an increase in apparent melting temperature relative to wild-type LEH from 50 to 85°C and a more than 250fold longer half-life, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 46°C and the enantiomeric excess is 80% in favor of (R,R)-cyclohexene-1,2-diol
-
T76K/L114V/I116V/F139V/L147F
-
site-directed mutagenesis, the mutant lacking the N-terminal mutations shows enantioselectivities of 82% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol, T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) is 45°C
-
T76K/L114V/I116V/N92K/F139V/L147F
-
site-directed mutagenesis, the mutant lacking the N-terminal mutations shows enantioselectivities of 83% enantiomeric excess in favor of (S,S)-cyclohexene-1,2-diol
-
Y53F
-
site-directed mutagenesis, QM/MM-optimized active structure of the enzyme mutant compared to wild-type
-
D80A
site-directed mutagenesis, mutation of the catalytic residue, inactive mutant
D82A
site-directed mutagenesis, mutation of the catalytic residue, inactive mutant
I80Y/L114V/I116V
-
site-directed mutagenesis, mutant SZ348, no or poor activity with cyclohexene-1,2-epoxide
L74F/M78F/I80F/L114V/I116V/F139V
-
site-directed mutagenesis, mutant SZ719
L74F/M78F/L103V/L114V/I116V/F139V/L147V
-
site-directed mutagenesis, mutant SZ92
L74F/M78V/I80V/L114F
-
site-directed mutagenesis, mutant SZ338, no activity with cyclohexene-1,2-epoxide
M32V/M78V/I80V/L114FM32V/M78V/I80V/L114F
-
site-directed mutagenesis, mutant SZ529
additional information
N55A
site-directed mutagenesis, QM/MM-optimized active structure of the enzyme mutant compared to wild-type
R99K
site-directed mutagenesis, QM/MM-optimized active structure of the enzyme mutant compared to wild-type
Y53F
site-directed mutagenesis, QM/MM-optimized active structure of the enzyme mutant compared to wild-type
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
additional information
mutations E45D, T76K, and N92K are located on or near the surface of LEH. It is likely that these mutations stabilize the protein by optimizing the distribution of charges on the enzyme surface, which is an established method of protein stabilization. Furthermore, S15D may form an ionic bond with A19K, thereby stabilizing a flexible N-terminal loop. Evolved thermostability-related mutations, structure-function relationships, overview
additional information
comparison of preparative resolution of (+)- and (-)-limonene oxide mixtures catalyzed by different LEHs under non-optimized conditions, optimization of the biocatalyzed processes to obtain enantiomerically pure isomers, overview
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
-
additional information
-
comparison of preparative resolution of (+)- and (-)-limonene oxide mixtures catalyzed by different LEHs under non-optimized conditions, optimization of the biocatalyzed processes to obtain enantiomerically pure isomers, overview
-
additional information
comparison of preparative resolution of (+)- and (-)-limonene oxide mixtures catalyzed by different LEHs under non-optimized conditions, optimization of the biocatalyzed processes, overview. The obtainment of the enantiomerically pure trans isomer, the increase of the reaction temperature to 50°C leads to an excellent resolution even at a substrate loading of 2 mol/l in reasonable reaction times
additional information
comparison of preparative resolution of (+)- and (-)-limonene oxide mixtures catalyzed by different LEHs under non-optimized conditions, optimization of the biocatalyzed processes, overview. The obtainment of the enantiomerically pure trans isomer, the increase of the reaction temperature to 50°C leads to an excellent resolution even at a substrate loading of 2 mol/l in reasonable reaction times
additional information
-
structural and computational insight into the catalytic mechanism of limonene epoxide hydrolase mutants in stereoselective transformations
additional information
comparison of preparative resolution of (+)- and (-)-limonene oxide mixtures catalyzed by different LEHs under non-optimized conditions, optimization of the biocatalyzed processes, overview. The obtainment of the cis isomer from the same (+)-limonene oxide mixture by Tomsk-LEH-catalyzed resolution is not significantly improved
additional information
comparison of preparative resolution of (+)- and (-)-limonene oxide mixtures catalyzed by different LEHs under non-optimized conditions, optimization of the biocatalyzed processes, overview. The obtainment of the cis isomer from the same (+)-limonene oxide mixture by Tomsk-LEH-catalyzed resolution is not significantly improved
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
41
wild-type LEH shows a thermostability of T50 30 (temperature at which 50% of enzyme activity is lost following a heat treatment for 30 min) = 41°C
additional information
R,R- and S,S-selective LEH variants (80-94% enantiomeric excess) show enhanced thermostability by 5-10°C compared to wild-type and still reasonable levels of activity
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
methyl-tert-butylether
enzyme maintains the catalytic activity
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzymes by nickel affinity and anion exchange chromatography, followed by ultrafiltration, gel filtration, and again ultrafiltration
recombinant soluble C-terminally His6-tagged enzyme from Escherichia coli strain 10G
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain 10G
recombinant expression of His-tagged wild-type and mutant enzymes
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
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
synthesis
construction of enzyme variants that produce enantiomeric diols from meso-epoxides. With substrate cis-stilbene oxide, enantiocomplementary mutants produce (S,S)- and (R,R)-stilbene diol with >97% enantiomeric excess. (R,R)-selective mutant M32L/L35G/I80W/L103V/F139L synthesizes (R,R)-stilbene diol with 98% conversion into diol, >99% ee
synthesis
construction of mutants as catalysts for preparing chiral N- and O-heterocycles with up to 99% conversion, and enantiomeric ratios up to 99:1. Baldwin-type cyclizations by the enzyme are enabled by reshaping the active-site environment that directs the distal RHN and HO-substituents to be intramolecular nucleophiles
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
-
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Van der Werf, M.J.; Orru, R.V.A.; Overkamp, K.M.; Swarts, H.J.; Ospiran, I.; Steinreiber, A.; de Bont, J.A.M.; Faber, K.
Substrate specificity and stereospecificity of limonene-1,2-epoxide hydrolase from Rhodococcus erythropolis DCL14; an enzyme showing sequential and enantioconvergent substrate conversion
Appl. Microbiol. Biotechnol.
52
380-385
1999
Rhodococcus erythropolis
-
brenda
Barbirato, F.; Verdoes, J.C.; de Bont, J.A.M.; van der Werf, M.J.
The Rhodococcus erythropolis DCL14 limonene-1,2-epoxide hydrolase gene encodes an enzyme belonging to a novel class of epoxide hydrolases
FEBS Lett.
438
293-296
1998
Rhodococcus erythropolis
brenda
Van der Werf, M.J.; Overkamp, K.M.; de Bont, J.A.M.
Limonene-1,2-epoxide hydrolase from Rhodococcus erythropolis DCL14 belongs to a novel class of epoxide hydrolases
J. Bacteriol.
180
5052-5057
1998
Rhodococcus erythropolis
brenda
Arand, M.; Hallberg, B.M.; Zou, J.; Bergfors, T.; Oesch, F.; Van Der Werf, M.J.; De Bont, J.A.M.; Jones, T.A.; Mowbray, S.L.
Structure of Rhodococcus erythropolis limonene-1,2-epoxide hydrolase reveals a novel active site
EMBO J.
22
2583-2592
2003
Rhodococcus erythropolis (Q9ZAG3), Rhodococcus erythropolis
brenda
Hopmann, K.H.; Hallberg, B.M.; Himo, F.
Catalytic mechanism of limonene epoxide hydrolase, a theoretical study
J. Am. Chem. Soc.
127
14339-14347
2005
Rhodococcus erythropolis (Q9ZAG3), Rhodococcus erythropolis DCL14 (Q9ZAG3), Rhodococcus erythropolis DCL14
brenda
Zheng, H.; Reetz, M.T.
Manipulating the stereoselectivity of limonene epoxide hydrolase by directed evolution based on iterative saturation mutagenesis
J. Am. Chem. Soc.
132
15744-15751
2010
Rhodococcus erythropolis, Rhodococcus erythropolis DCL 14
brenda
Molina, G.; Bution, M.L.; Bicas, J.L.; Dolder, M.A.; Pastore, G.M.
Comparative study of the bioconversion process using R-(+)- and S-(-)-limonene as substrates for Fusarium oxysporum 152B
Food Chem.
174
606-613
2015
Fusarium oxysporum, Fusarium oxysporum 152B
brenda
Li, G.; Zhang, H.; Sun, Z.; Liu, X.; Reetz, M.
Multiparameter optimization in directed evolution engineering thermostability, enantioselectivity, and activity of an epoxide hydrolase
ACS Catal.
6
3679-3687
2016
Rhodococcus erythropolis (Q9ZAG3), Rhodococcus erythropolis DCL14 (Q9ZAG3)
-
brenda
Rinaldi, S.; Van Der Kamp, M.; Ranaghan, K.; Mulholland, A.; Colombo, G.
Understanding complex mechanisms of enzyme reactivity the case of limonene-1,2-epoxide hydrolases
ACS Catal.
8
5698-5707
2018
Rhodococcus erythropolis (Q9ZAG3), Rhodococcus erythropolis DCL14 (Q9ZAG3)
-
brenda
Hou, Q.Q.; Sheng, X.; Wang, J.H.; Liu, Y.J.; Liu, C.B.
QM/MM study of the mechanism of enzymatic limonene 1,2-epoxide hydrolysis
Biochim. Biophys. Acta
1824
263-268
2012
Rhodococcus erythropolis (Q9ZAG3), Rhodococcus erythropolis DCL14 (Q9ZAG3)
brenda
Ferrandi, E.; Marchesi, C.; Annovazzi, C.; Riva, S.; Monti, D.; Wohlgemuth, R.
Efficient epoxide hydrolase catalyzed resolutions of (+)- and (-)-cis/trans-limonene oxides
ChemCatChem
7
3171-3178
2015
Rhodococcus erythropolis (Q9ZAG3), Rhodococcus erythropolis DCL14 (Q9ZAG3), uncultured organism (A0A0G3IAY2), uncultured organism (A0A0G3ICV8)
-
brenda
Ferrandi, E.E.; Sayer, C.; Isupov, M.N.; Annovazzi, C.; Marchesi, C.; Iacobone, G.; Peng, X.; Bonch-Osmolovskaya, E.; Wohlgemuth, R.; Littlechild, J.A.; Monti, D.
Discovery and characterization of thermophilic limonene-1,2-epoxide hydrolases from hot spring metagenomic libraries
FEBS J.
282
2879-2894
2015
Rhodococcus erythropolis (Q9ZAG3), Rhodococcus erythropolis, uncultured organism (A0A0G3IAY2), uncultured organism (A0A0G3ICV8)
brenda
Sun, Z.; Wu, L.; Bocola, M.; Chan, H.C.S.; Lonsdale, R.; Kong, X.D.; Yuan, S.; Zhou, J.; Reetz, M.T.
Structural and computational insight into the catalytic mechanism of limonene epoxide hydrolase mutants in stereoselective transformations
J. Am. Chem. Soc.
140
310-318
2018
uncultured organism
brenda
Bassanini, I.; Ferrandi, E.; Monti, D.; Riva, S.
Studies on the catalytic promiscuity of limonene epoxide hydrolases in the non-hydrolytic ring opening of 1,2-epoxides
ChemBioChem
21
1868-1874
2020
Rhodococcus erythropolis (Q9ZAG3)
brenda
Arabnejad, H.; Bombino, E.; Colpa, D.I.; Jekel, P.A.; Trajkovic, M.; Wijma, H.J.; Janssen, D.B.
Computational design of enantiocomplementary epoxide hydrolases for asymmetric synthesis of aliphatic and aromatic diols
ChemBioChem
21
1893-1904
2020
Rhodococcus erythropolis (Q9ZAG3)
brenda
Li, J.K.; Qu, G.; Li, X.; Tian, Y.; Cui, C.; Zhang, F.G.; Zhang, W.; Ma, J.A.; Reetz, M.T.; Sun, Z.
Rational enzyme design for enabling biocatalytic Baldwin cyclization and asymmetric synthesis of chiral heterocycles
Nat. Commun.
13
7813
2022
Rhodococcus erythropolis (Q9ZAG3)
brenda
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