Systematic Investigation ofSaccharomycescerevisiaeEnzymes Catalyzing Carbonyl Reductions

Abstract

Eighteen key reductases from baker's yeast (Saccharomyces cerevisiae) have been overproduced in Escherichia coli as glutathione S-transferase fusion proteins. A representative set of alpha- and beta-keto esters was tested as substrates (11 total) for each purified fusion protein. The stereoselectivities of beta-keto ester reductions depended both on the identity of the enzyme and the substrate structure, and some reductases yielded both L- and D-alcohols with high stereoselectivities. While alpha-keto esters were generally reduced with lower enantioselectivities, it was possible in all but one case to identify pairs of yeast reductases that delivered both alcohol antipodes in optically pure form. Taken together, the results demonstrate not only that individual yeast reductases can be used to supply important chiral building blocks, but that GST-fusion proteins allow rapid identification of synthetically useful biocatalysts (along with their corresponding genes).

Bioreductive preparation of ACE inhibitors precursor (R)-2-hydroxy-4-phenylbutanoate esters: Recent advances and future perspectives

“…The shotgun library of OPBE [53]. Through genome hunting, Ypr1p and YOL151w were identified from Saccharomyces cerevisiae with 87% and 99% ee (R), respectively [54,55]. NcCR was cloned from Neurospora crassa with 1.6 U mg −1 specific activity and 80% ee (R) [56].…”

Recent trends in whole cell and isolated enzymes in enantioselective synthesis

“…339 A further advantage of ketoreductase technology is the rapidity by which these screens can be performed. 339 An overview of the latest achievements in the field of asymmetric ketoreductase-mediated carbonyl reductions can be found in recent reviews. 29,66,327,[340][341][342] …”

“…338 When Stewart et al examined the genome of Baker's yeast the results indicated the potential for 50 ketoreductases of which 19 were over expressed, isolated, and studied. 339 The novel isolated ketoreductases reduced a wide range of ketones and produced both enantiomers of most products depending on the ketoreductase employed ( Figure 19 Production of at least two of the four possible 3-chloro-2-hydroxy ester diastereomers with high optical purities…”

Stereochemistry of Furfural Reduction by a Saccharomyces cerevisiae Aldehyde Reductase That Contributes to In Situ Furfural Detoxification

“…The use of SDRs as chiral induction agents for the generation of reduced materials with high enantiomeric excess has been the subject of many studies (15,31). Specifically, Ari1p (YGL157Wp) has previously been shown to be capable of reducing ␣-and ␤-keto esters with high enantioselectivity (18,19) and t-butyl 6-chloro-3,5-dioxohexanoate with high enantioselectivity and regiospecificity (38). Within the data reported, the Ari1p-catalyzed reduction of ␤-keto esters and ␣-chloro-␤-keto esters with substituents ranging in size from methyl to benzyl occurs from the re face regardless of substituent (18,19).…”

“…Specifically, Ari1p (YGL157Wp) has previously been shown to be capable of reducing ␣-and ␤-keto esters with high enantioselectivity (18,19) and t-butyl 6-chloro-3,5-dioxohexanoate with high enantioselectivity and regiospecificity (38). Within the data reported, the Ari1p-catalyzed reduction of ␤-keto esters and ␣-chloro-␤-keto esters with substituents ranging in size from methyl to benzyl occurs from the re face regardless of substituent (18,19). Variations in the structure, including a ␦-branched substrate and a phenyl-substituted ketone, were poorly reduced, and no stereochemical data were reported (18,19); however, in other yeast reductases the change in substrate size was sufficient to provide the opposite stereochemical product (19), suggesting that Ari1p may not tolerate certain bulky substituents according to the large and small binding pocket model (36).…”