Optimization of the solvent-tolerant Pseudomonas putida S12 as host for the production of p-coumarate from glucose

Appl Microbiol Biotechnol. 2007 Mar;74(3):617-24. doi: 10.1007/s00253-006-0703-0. Epub 2006 Nov 17.

Abstract

A Pseudomonas putida S12 strain was constructed that is able to convert glucose to p-coumarate via the central metabolite L: -tyrosine. Efficient production was hampered by product degradation, limited cellular L: -tyrosine availability, and formation of the by-product cinnamate via L: -phenylalanine. The production host was optimized by inactivation of fcs, the gene encoding the first enzyme in the p-coumarate degradation pathway in P. putida, followed by construction of a phenylalanine-auxotrophic mutant. These steps resulted in a P. putida S12 strain that showed dramatically enhanced production characteristics with controlled L: -phenylalanine feeding. During fed-batch cultivation, 10 mM (1.7 g l(-1)) of p-coumarate was produced from glucose with a yield of 3.8 Cmol% and a molar ratio of p-coumarate to cinnamate of 85:1.

MeSH terms

  • Cinnamates / metabolism
  • Coenzyme A Ligases / genetics
  • Coumaric Acids / metabolism*
  • Fermentation
  • Gene Deletion
  • Glucose / metabolism*
  • Phenylalanine / biosynthesis
  • Phenylalanine / genetics
  • Propionates
  • Pseudomonas putida / genetics
  • Pseudomonas putida / metabolism*
  • Tyrosine / metabolism

Substances

  • Cinnamates
  • Coumaric Acids
  • Propionates
  • cinnamic acid
  • Tyrosine
  • Phenylalanine
  • Coenzyme A Ligases
  • feruloyl-coenzyme A synthetase
  • p-coumaric acid
  • Glucose