Stereoselective Bioreduction of Ethyl 3-Oxo-3-(2-Thienyl) Propanoate Using the Short-Chain Dehydrogenase/Reductase Ch KRED12

J Microbiol Biotechnol. 2019 Nov 28;29(11):1769-1776. doi: 10.4014/jmb.1805.04058.

Abstract

Ethyl (S)-3-hydroxy-3-(2-thienyl)propanoate((S)-HEES)acts as a key chiral intermediate for the blockbuster antidepressant drug duloxetine, which canbe achieved viathe stereoselective bioreduction ofethyl 3-oxo-3-(2-thienyl) propanoate (KEES) that containsa 3-oxoacyl structure.The sequences of the short-chain dehydrogenase/reductases from Chryseobacterium sp. CA49 were analyzed, and the putative3-oxoacyl-acyl-carrier-protein reductase, ChKRED12, was able to stereoselectivelycatalyze theNADPH-dependent reduction to produce (S)-HEES.The reductase activity of ChKRED12 towardsothersubstrates with 3-oxoacyl structure were confirmed with excellent stereoselectivity (>99% enantiomeric excess) in most cases. When coupled with a cofactor recycling system using glucose dehydrogenase, the ChKRED12 was able to catalyze the complete conversion of 100 g/l KEES within 12h, yielding the enantiopure product with >99% ee, showing a remarkable potential to produce (S)-HEES.

Keywords: 3-oxoacyl-acyl-carrier-protein reductase; Bioreduction; duloxetine; short-chain dehydrogenase.

MeSH terms

  • 3-Oxoacyl-(Acyl-Carrier-Protein) Reductase / chemistry
  • 3-Oxoacyl-(Acyl-Carrier-Protein) Reductase / genetics
  • 3-Oxoacyl-(Acyl-Carrier-Protein) Reductase / metabolism*
  • Amino Acid Sequence
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Catalysis
  • Chryseobacterium / enzymology
  • Chryseobacterium / genetics
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Glucose 1-Dehydrogenase / metabolism
  • Kinetics
  • Oxidation-Reduction
  • Propionates / chemistry
  • Propionates / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Short Chain Dehydrogenase-Reductases / chemistry
  • Short Chain Dehydrogenase-Reductases / genetics
  • Short Chain Dehydrogenase-Reductases / metabolism*
  • Stereoisomerism
  • Substrate Specificity

Substances

  • Bacterial Proteins
  • Propionates
  • Recombinant Proteins
  • Short Chain Dehydrogenase-Reductases
  • 3-Oxoacyl-(Acyl-Carrier-Protein) Reductase
  • Glucose 1-Dehydrogenase