Asymmetric bioreduction of activated alkenes by a novel isolate of Achromobacter species producing enoate reductase

Abstract

The strain Achromobacter sp. JA81, which produced enoate reductase, was applied in the asymmetric reduction of activated alkenes. The strain could catalyze the bioreduction of alkenes to form enantiopure (R)-β-aryl-β-cyano-propanoic acids, a precursor of (R)-γ-amino butyric acids, including the pharmaceutically active enantiomer of the chiral drug (R)-baclofen with excellent enantioselectivity. It could catalyze as well the stereoselective bioreduction of other activated alkenes such as cyclic imides, β-nitro acrylates, and nitro-alkenes with up to >99 % ee and >99 % conversion. The draft genome sequencing of JA81 revealed six putative old yellow enzyme homologies, and the transcription of one of them, Achr-OYE3, was detected using reverse transcription polymerase chain reaction. The recombinant Escherichia coli expressing Achr-OYE3 displayed enoate reductase activity toward (Z)-3-cyano-3-phenyl-propenoic acid (2a).

An enoate reductase Achr-OYE4 from Achromobacter sp. JA81: characterization and application in asymmetric bioreduction of C=C bonds

“…JA81, Achr-OYE3 was the only one whose transcription was detected in the cells cultivated under standard conditions (Liu et al 2012). Further investigation using cells cultivated in the presence of possible inducers also showed the absence of transcription of Achr-OYE4 despite transcription of some of the other OYE homologies (data not shown).…”

“…However, no product formation was observed for the corresponding α,β-unsaturated acid, (Z)-3-cyano-3-phenylacrylic acid, which had been accepted as a substrate by the native strain of Achromobacter sp. JA81 (Liu et al 2012).…”

“…JA81 is a strain recently isolated from soil and is recognized for its ability to catalyze the enantioselective reduction of β-aryl-β-cyano-α,β-unsaturated carboxylic acid to (R)-β-aryl-β-cyano-carboxylic acid (Liu et al 2012). The strain can also catalyze the bioreduction of other commonly investigated activated alkenes such as cyclic imides, β-nitro acrylates, and nitro-alkenes with up to >99 % ee and >99 % conversion (Liu et al 2012).…”

“…JA81 is a strain recently isolated from soil and is recognized for its ability to catalyze the enantioselective reduction of β-aryl-β-cyano-α,β-unsaturated carboxylic acid to (R)-β-aryl-β-cyano-carboxylic acid (Liu et al 2012). The strain can also catalyze the bioreduction of other commonly investigated activated alkenes such as cyclic imides, β-nitro acrylates, and nitro-alkenes with up to >99 % ee and >99 % conversion (Liu et al 2012). The coding sequences of six OYE homologues, designated as Achr-OYE1 to Achr-OYE6, have been identified from its draft genome sequence, and the transcription of Achr-OYE3 has been detected using standard cultivation conditions (Liu et al 2012).…”

“…The strain can also catalyze the bioreduction of other commonly investigated activated alkenes such as cyclic imides, β-nitro acrylates, and nitro-alkenes with up to >99 % ee and >99 % conversion (Liu et al 2012). The coding sequences of six OYE homologues, designated as Achr-OYE1 to Achr-OYE6, have been identified from its draft genome sequence, and the transcription of Achr-OYE3 has been detected using standard cultivation conditions (Liu et al 2012).…”