Two Enantiocomplementary Ephedrine Dehydrogenases from Arthrobacter sp. TS-15 with Broad Substrate Specificity

Gideon James Grogan, Tarek Shanati, Cameron Lockie, Lilian Beloti, Marion Ansorge-Schumacher

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The recently identified pseudoephedrine and ephedrine dehydrogenases from Arthrobacter sp. TS-15, PseDH and EDH, are NADH-dependent members of the oxidoreductase superfamily of short-chain dehydrogenases/reductases (SDRs). They are specific for the enantioselective oxidation of (+)-(S) N-(pseudo)ephedrine and (-)-(R) N-(pseudo)ephedrine, respectively. Anti-Prelog stereospecific PseDH and Prelog-specific EDH catalyse the regio- and enantiospecific reduction of 1-phenyl-1,2-propanedione to (S)-phenylacetylcarbinol and (R)-phenylacetylcarbinol with full conversion and enantiomeric excess of >99%. Moreover, they perform the reduction of a wide range of aryl aliphatic carbonyl compounds, including keto amines, ketoesters and haloketones, to the corresponding enantiopure alcohols. The highest stability of PseDH and EDH was determined to be at a pH range of 6.0-8.0 and 7.5-8.5, respectively. PseDH was more stable than EDH at 25 °C, with half-lives of 279 h and 38 h respectively. However, EDH is more stable at 40 °C with a two-fold greater half-life than at 25 °C. The crystal structure of the PseDH-NAD+ complex, refined to a resolution of 1.83 Å, revealed a tetrameric structure, which was confirmed by solution studies. A model of the active site in complex with NAD+ and 1-phenyl-1,2-propanedione suggested key roles for S143 and W152 in recognition of the substrate and positioning for the reduction reaction. The wide substrate spectrum of these dehydrogenases, combined with their regio- and enantioselectivity, suggests high potential for the industrial production of valuable chiral compounds.
Original languageEnglish
Article number6202–6211
Number of pages10
JournalACS Catalysis
Issue number7
Early online date29 May 2019
Publication statusPublished - 5 Jul 2019

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