Applied Microbiology and Biotechnology

, Volume 74, Issue 3, pp 617–624 | Cite as

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

  • Karin Nijkamp
  • R. G. Maaike Westerhof
  • Hendrik Ballerstedt
  • Jan A. M. de Bont
  • Jan Wery
Applied Genetics and Molecular Biotechnology

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.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Karin Nijkamp
    • 1
  • R. G. Maaike Westerhof
    • 1
  • Hendrik Ballerstedt
    • 1
  • Jan A. M. de Bont
    • 1
    • 2
  • Jan Wery
    • 1
  1. 1.Business Unit, Food and Biotechnology InnovationsTNO Quality of LifeApeldoornThe Netherlands
  2. 2.Faculty of Applied SciencesDelft University of TechnologyDelftThe Netherlands

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