Optimization of the solvent-tolerantPseudomonas putidaS12 as host for the production ofp-coumarate from glucose

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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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