Optimization of the solvent-tolerant Pseudomonas putida S12 as host for the production of p-coumarate from glucose
article
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. © 2006 Springer-Verlag.
Topics
BiotechnologyDegradationFeedingGenesGenetic engineeringGlucoseMutagenesisOptimizationProduction controlSolventsCellular L-tyrosineMolar ratioP-coumarateSolvent-tolerant Pseudomonas putidaBacteriaCinnamic acidGlucosePara coumaric acidPhenylalanineTyrosineAmino acidBacteriumBiodegradationDegradationEnzymeGlucoseMetaboliteOptimizationOrganic acidControlled studyFeedingHostMutantNonhumanPseudomonas putidaCinnamatesCoenzyme A LigasesCoumaric AcidsFermentationGene DeletionGlucosePhenylalaninePseudomonas putidaTyrosinePseudomonas putida
TNO Identifier
239860
ISSN
01757598
Source
Applied Microbiology and Biotechnology, 74(3), pp. 617-624.
Pages
617-624
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