Systematic investigation of Saccharomyces cerevisiae enzymes catalyzing carbonyl reductions.
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).
Contribution from the Department of Chemistry, University of Florida, Gainesville, FL 32611, USA.
Iwona A
IA
Kaluzna
Tomoko
T
Matsuda
Aileen K
AK
Sewell
Jon D
JD
Stewart
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D004952:Esters / Q000737:chemistry / Q000378:metabolism*; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D007659:Ketones / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity
D004952:Esters; D007659:Ketones; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D000429:Alcohol Oxidoreductases; D005982:Glutathione Transferase