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Asymmetric Reduction of α-Keto Esters with Thermus thermophilus NADH-Dependent Carbonyl Reductase using Glucose Dehydrogenase and Alcohol Dehydrogenase for Cofactor Regeneration

✍ Scribed by Angela Pennacchio; Assunta Giordano; Mosè Rossi; Carlo A. Raia


Publisher
John Wiley and Sons
Year
2011
Tongue
English
Weight
155 KB
Volume
2011
Category
Article
ISSN
1434-193X

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✦ Synopsis


Abstract

The enantioselective synthesis of methyl (R)‐mandelate and methyl (R)‐o‐chloromandelate was investigated using an NADH‐dependent carbonyl reductase from Thermus thermophilus (TtADH) and, separately, archaeal glucose dehydrogenase and Bacillus stearothermophilus alcohol dehydrogenase (BsADH) for NADH regeneration. Optimal reaction times and substrate concentrations in the absence and presence of organic solvents were determined. The enantiofacial selectivity of TtADH was shown to be inversely proportional to the hydrophobicity of the short‐chain linear alcohols employed as co‐substrates of the bacillar ADH. The bioreduction of methyl benzoylformate yielded the (R)‐alcohol with a 77 % yield (ee = 96 %) using glucose dehydrogenase and glucose, and 81 % yield (ee = 94 %) applying BsADH and ethanol. The bioreduction of methyl o‐chlorobenzoylformate yielded the halogenated (R)‐alcohol with 95 % and 92 % ee, and 62 % and 78 % yield using glucose dehydrogenase and BsADH, respectively.