Substrate specificity and stereospecificity of limonene-1,2-epoxide hydrolase from Rhodococcus erythropolis DCL14; an enzyme showing sequential and enantioconvergent substrate conversion

Werf, M.J. van der; Orru, Romano V. A.; Overkamp, K.M.; Swarts, Hans J. M.; Osprian, Ingrid; Steinreiber, Andreas; Bont, J.A.M. de; Faber, Kurt

doi:10.1007/s002530051535

Cited by 64 publications

(52 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This stereochemical outcome is interestingly in line with the results previously observed with the most reactive enantiomer of limonene-1,2-epoxide [17] as well as for 1-methylcyclohexene oxide. [18] Optimisation of the Substrate Concentration…”

supporting

confidence: 91%

Epoxide Hydrolase‐Catalysed Kinetic Resolution of a Spiroepoxide, a Key Building Block of Various 11‐Heterosteroids

et al. 2007

View full text Add to dashboard Cite

Two microbial epoxide hydrolases -i.e., Aspergillus niger (AnEH) and Rhodococcus erythropolis (the so-called "Limonene EH": LEH) were used to achieve, for the first time, the biocatalysed hydrolytic kinetic resolution (BHKR) of spiroepoxide rac-1. This compound is a strategic key building block allowing the synthesis of 11-heterosteroids. Interestingly enough, the two enzymes exhibited opposite and therefore complementary enantioselectivity allowing us to isolate the residual (R,R)-1 (from AnEH) and the residual (S,S)-1 (from LEH) in nearly enantiopure forms (> 98 %). Their absolute configurations were determined by X-ray crystallography. An opposite regioselectivity of the oxirane ring opening for both enantiomers of substrate 1, determined using H 2 18O labelling and chiral GC-MS analysis, was also observed, corresponding to an attack at the less substituted carbon atom using AnEH, and at the most substituted carbon atom using LEH. A chemical process-improving methodology was also developed. This allowed us to obtain both enantiomers of the substrate in high enantiomeric purity (99 %) and optimised quantity. In the case of the AnEH, the use of a biphasic (water/isooctane) reaction medium allowed us to increase the global substrate concentration up to 200 g/ L. The preparation of both enantiomers of 1 clearly paves the way to the preparative scale synthesis and biochemical evaluation of the corresponding 11-heterosteroid enantiomers.

show abstract

supporting

confidence: 91%

Epoxide Hydrolase‐Catalysed Kinetic Resolution of a Spiroepoxide, a Key Building Block of Various 11‐Heterosteroids

et al. 2007

View full text Add to dashboard Cite

show abstract

“…Although the enantioselectivity of this class of enzymes has been shown for the rigid substrate limonene 1,2-epoxide, 16 the phenomenon is rather unexpected with 9,10-epoxystearic acid, since the groups that could direct such selectivity are quite distant from the catalytic center ( Figure 1). …”

Section: Discussionmentioning

confidence: 90%

Structure of an Atypical Epoxide Hydrolase from Mycobacterium tuberculosis Gives Insights into its Function

Johansson

Unge

Cronin

et al. 2005

Journal of Molecular Biology

View full text Add to dashboard Cite

“…The enzyme with the most interesting (unusual) activity towards these substrates is the limonene-1,2-epoxide hydrolase (LEH) of Rhodococcus erythropolis DCL14. This enzyme, which is not a α/β-hydrolase fold enzyme, gives, via an unusual acid-catalysed mechanism, enantioconvergent hydrolysis of both sets of epoxide isomers to yield from the (4R) epoxide isomers only the (1S,2S,4R) diol and from the (4S) epoxide isomers only the (1R,2R,4S diol) [63]. Because conversion of the cis/trans diastereomer mixtures is sequential the enzyme can also be used to obtain in the case of the (4R)-epoxides the remaining trans-diastereomer in 99% purity [64], while in the case of the (4S)-epoxides the unreacted cis-diastereomer was recovered [63].…”

Section: Types III and V: 22-disubstituted Epoxides And Trisubstitutedmentioning

confidence: 99%

“…(11). Conversion of the four stereoisomers of limonene-1,2-epoxide into diaxial limonene-1,2-diol by purified LEH from Rhodococcus erythropolis [63]. driving low levels of gene expression.…”

Section: Recombinant Epoxide Hydrolasesmentioning

confidence: 99%

Diversity of Epoxide Hydrolase Biocatalysts

Labuschagne¹

2006

COC

View full text Add to dashboard Cite

Chiral epoxides and their derivatives are versatile intermediates for the synthesis of enantiopure organic chemicals. Chiral epoxides can be obtained through asymmetric epoxidation using either chemical catalysts or epoxidizing enzymes such as monooxygenases or chloroperoxidases. Alternatively hydrolytic kinetic resolution of racemic epoxides can yield a mixture containing, in an ideal situation, a single enantiomer of the remaining epoxide and a single enantiomer of the formed diol. Occasionally catalysts with complementary enantioselectivities and opposite regioselectivities can give through enantioconvergent hydrolysis one diol enantiomer in > 90% yield. Hydrolyses can also be performed with either metal catalysts or biological catalysts, mainly epoxide hydrolases. Epoxide hydrolases show great promise for the preparative scale hydrolysis of epoxides, because they are easy-to-use biocatalysts that accept a wide range of substrates, that do not require co-factors and that show improved activity when the substrate is present as a second water immiscible phase. X-ray structures for one fungal, one bacterial and two mammalian epoxide hydrolases are available as well as gene sequences for more than ninety five epoxide hydrolases. Expression systems in Escherichia coli and other hosts have also been developed and different assays suitable for high throughput screening have been developed and tested. The latter developments also made possible the first attempts at catalyst improvement through site directed mutagenesis and directed evolution. This review will give a comparative overview of the different types of epoxide hydrolases, which recently gave promising results for hydrolytic kinetic or enantioconvergent resolution of different types of epoxides and of results obtained with various recombinant epoxide hydrolases.

show abstract

Substrate specificity and stereospecificity of limonene-1,2-epoxide hydrolase from Rhodococcus erythropolis DCL14; an enzyme showing sequential and enantioconvergent substrate conversion

Cited by 64 publications

References 36 publications

Epoxide Hydrolase‐Catalysed Kinetic Resolution of a Spiroepoxide, a Key Building Block of Various 11‐Heterosteroids

Epoxide Hydrolase‐Catalysed Kinetic Resolution of a Spiroepoxide, a Key Building Block of Various 11‐Heterosteroids

Structure of an Atypical Epoxide Hydrolase from Mycobacterium tuberculosis Gives Insights into its Function

Diversity of Epoxide Hydrolase Biocatalysts

Contact Info

Product

Resources

About