Enzymatic preparation of D-phenyllactic acid at high space-time yield with a novel phenylpyruvate reductase identified from Lactobacillus sp. CGMCC 9967

J Biotechnol. 2016 Mar 20:222:29-37. doi: 10.1016/j.jbiotec.2015.12.011. Epub 2015 Dec 19.

Abstract

An NADH-dependent phenylpyruvate reductase (LaPPR) was identified through screening the shotgun library of Lactobacillus sp. CGMCC 9967. It belongs to D-3-phosphoglycerate dehydrogenase (PGDH) subfamily of 2-hydroxy acid dehydrogenase superfamily. LaPPR was stable at pH 6.5 and 30 °C, with a half-life of 152 h. LaPPR has a substrate preference towards aromatic to aliphatic keto acids, and various keto acids could be reduced into D-hydroxy acids with excellent enantioselectivity (>99%). By construction the coexpression system with glucose dehydrogenase, as much as 100 g L(-1) phenylpyruvic acid was asymmetrically reduced into D-phenyllactic acid with 91.3% isolation yield and 243 g L(-1) d(-1) productivity. The results suggest that LaPPR is a promising biocatalyst for the efficient synthesis of optically pure D-phenyllactic acid.

Keywords: Asymmetric reduction; Lactobacillus sp. CGMCC 9976; Phenylpyruvate reductase; d-phenyllactic acid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Lactates / metabolism*
  • Lactobacillus / enzymology*
  • Lactobacillus / genetics*
  • Models, Molecular
  • Oxidoreductases / chemistry
  • Oxidoreductases / genetics
  • Oxidoreductases / metabolism

Substances

  • Bacterial Proteins
  • Lactates
  • Oxidoreductases
  • phenylpyruvate oxidase