Heterologous expression of a novel d‑lactate dehydrogenase from Lactobacillus sp. ZX1 and its application for d‑phenyllactic acid production

Int J Biol Macromol. 2018 Nov:119:1171-1178. doi: 10.1016/j.ijbiomac.2018.08.036. Epub 2018 Aug 10.

Abstract

d‑Phenyllactic acid (d‑PLA) shows great potential for biopreservative production owning to its anti-microbial activity. In this study, strain ZX1, which could inhibit the growth of other microbes was isolated and identified as Lactobacillus genus. Strain ZX1 could produce d‑PLA with 0.16 g·L-1. Furthermore, a novel d‑lactate dehydrogenase gene was identified and expressed in Escherichia coli BL21 (DE3) with the specific activity of 71.64 U·mg-1 protein under the optimal temperature and pH of 35 °C and 6.2. The kinetic constants for Kcat, Km, Vmax were 10.71 s-1, 2.356 mM and 11.27 μM·mg-1·min-1 for phenylpyruvic acid (PPA), respectively. 18.21 g·L-1d‑PLA from PPA with yield of 90.49% and productivity of 2.49 g·L-1·h-1 was obtained by the recombinant E. coli BL21 harboring d‑LDH, indicating that engineered E. coli BL21 (d‑LDH) has excellent potential in commercial d‑PLA production.

Keywords: Bacteriostasis; Lactobacillus sp.; Whole cell catalysis; d‑Lactate dehydrogenase; d‑Phenyllactic acid.

MeSH terms

  • Amino Acid Sequence
  • Biotransformation
  • Cloning, Molecular
  • Enzyme Stability
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Genetic Engineering*
  • Kinetics
  • Lactate Dehydrogenases / chemistry
  • Lactate Dehydrogenases / genetics*
  • Lactate Dehydrogenases / metabolism*
  • Lactates / metabolism*
  • Lactobacillus / enzymology*
  • Lactobacillus / genetics
  • Metals / pharmacology
  • Temperature

Substances

  • Lactates
  • Metals
  • 3-phenyllactic acid
  • Lactate Dehydrogenases
  • D-lactate dehydrogenase