A Single Point Mutation Reverses the Enantiopreference of Thermoanaerobacter ethanolicus Secondary Alcohol Dehydrogenase

Abstract

The asymmetric reduction of benzylic and heteroaryl ketones to the corresponding (R)‐alcohols using I86A Thermoanaerobacter ethanolicus alcohol dehydrogenase (I86A TeSADH) is described. This single amino acid mutation not only makes the active site of I86A TeSADH able to accommodate more sterically demanding substituents than those accommodated by wild‐type TeSADH, but it also reverses the substrate stereospecificity of TeSADH.

A new dehydrogenase from Clostridium acetobutylicum for asymmetric synthesis: dynamic reductive kinetic resolution entry into the Taxotère side chain

“…Biocatalytic synthesis with purified enzymes, including aldolases, 1 epoxide hydrolases, 2 Baeyer-Villigerases, 3 amine oxidases, 4 lipases, 5 and alcohol dehydrogenases, 6 is emerging as a viable alternative to traditional asymmetric synthesis. The recent transaminase-based asymmetric synthesis of the Januvia™ core by the Merck-Codexis team highlights the viability of such approaches, and was recognized with the 2010 Presidential Green Challenge Award.…”

Semi–Rational Engineering a Carbonyl Reductase for the Enantioselective Reduction of β-Amino Ketones

“…Although combinational mutations of other enzymes have successfully resulted in reversal in stereoselectivity, 6d,f,11 there are few examples of reversed enzyme enantiopreference by one single-point mutation in the literature, 12,13 not to mention the carbonyl reductase. 5c, 14 This study not only provides valuable information for our understanding of how to alter the enantioselectivity of this enzyme but also offers a promising starting point for further addressing the challenge in the reduction of sterically hindered β-amino ketones such as 1a and 1b to meet the needs of synthetic application.…”

Two‐Enzyme Hydrogen‐Borrowing Amination of Alcohols Enabled by a Cofactor‐Switched Alcohol Dehydrogenase

“…A single NADH‐dependent racemase has been described, but the substrate scope and reaction parameters have not been explored . Interestingly, Musa and co‐workers recently reported engineered variants of the NAD(P)H‐dependent ADH from Thermoanerobacter ethanolicus (TeSADH) that showed low enantioselectivity in the reduction of phenylacetone . The TeSADH meets many of the required criteria with respect to selectivity, substrate scope, stability, and ease of handling.…”