Asymmetric Bioreduction of Activated C=C Bonds Using Zymomonas mobilis NCR Enoate Reductase and Old Yellow Enzymes OYE 1–3 from Yeasts

Abstract

The asymmetric bioreduction of C=C-bonds bearing an electron-withdrawing group, such as an aldehyde, ketone, imide, nitro, carboxylic acid, or ester moiety by a novel enoate reductase from Zymomonas mobilis and Old Yellow Enzymes OYE 1-3 from yeasts furnished the corresponding saturated products in up to Ͼ99 % ee. Depending on the substrate type, stereocontrol was achieved by variation of the substrate

Activated α,β‐Unsaturated Aldehydes as Substrate of Dihydroxyacetone Phosphate (DHAP)‐Dependent Aldolases in the Context of a Multienzyme System

“…The enzyme was inactive for more sterically hindered substrates such as 3-phenylcyclohexene-1-one. Similar trends have also been seen with other OYEs, [2][3][4] which is consistent with the structural similarity of the enzymes within the family. However, recent studies show that directed evolution can be used to extend the substrate specificity of OYEs towards more sterically hindered substrates.…”

“…This unexpected change of enantioselectivity for formation of 2-methylcyclopentanone 17b has also been reported for other OYEs, although no explanation for this observation was given. [2][3][4] In light of recent work by Stewart et al [5] it is possible that substrate 17a may bind in a 'flipped' orientation, while maintaining an optimal Cβ-FMN N5 distance and angle (105°), [21] similar to that seen for OYE1 mutant W116I. [5] PETNR proved to be highly efficient and highly stereoselective in the reduction of 2-methylmaleimide derivatives 1a-b giving the respective (2R)-products in quantitative yields and high enantioselectivities (>99% ee, Table 2 and Scheme 1c).…”

Identification of enzymes responsible for the reduction of geraniol to citronellol

“…This activity was previously observed for the recombinant OYE2. 6 All the recombinant enzymes were inactive to methyl crotonate and methyl transcinnamate based on GC-MS analyses (data not shown).…”

“…Recently, enzymes from the old yellow enzyme (OYE) family have been discovered to catalyze the asymmetric reduction of activated alkenes, creating up to two chiral centers and, thus, showing a great potential in the industrial reduction of many commercially useful substrates. [3][4][5][6] The OYEs are characterized by high stereospecificity, strict regioselectivity, and rather broad substrate specificity. Fungi, particularly various yeasts, and bacteria were prominently used in whole-cell biotransformations of activated alkenes; whereas plant cells have only a minor part in the field.…”

“…To avoid these problems, isolated and/or cloned enzymes have gained growing interest in biocatalytic applications. Successful attempts in the chemo-and stereoselective reduction of activated alkenes were reported by using cloned yeast OYE1 from Saccharomyces carlsbergensis, [5][6][7] , OYE2 and OYE3 from S. cerevisiae, [6][7][8] CmOYE from Candida macedoniensis; 9 and bacterial OYE homologues YqjM from Bacillus subtilis, NCR from Zymomonas mobilis; 6,10 and plant OYE homologues LeOPR1 and LeOPR3 from Lycopersicon esculentum (tomato). 10,11 The plant 12-oxophytodienoate reductase (OPR) is involved in the reduction of (9S,13S)-12-oxophytodienoate to the corresponding cyclopentanoic acid OPC-8:0 for jasmonate biosynthesis.…”