title: Optimization of the solvent-tolerant Pseudomonas putida S12 as host for the production of p-coumarate from glucose.
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.
authors.affiliation: Business Unit, Food and Biotechnology Innovations, TNO Quality of Life, P.O. Box 342, 7300 AH, Apeldoorn, The Netherlands. karin.nijkamp@tno.nl
authors.forename: Karin
authors.initials: K
authors.lastname: Nijkamp
authors.forename: R G Maaike
authors.initials: RG
authors.lastname: Westerhof
authors.forename: Hendrik
authors.initials: H
authors.lastname: Ballerstedt
authors.forename: Jan A M
authors.initials: JA
authors.lastname: de Bont
authors.forename: Jan
authors.initials: J
authors.lastname: Wery
mesh_terms: D002934:Cinnamates / Q000378:metabolism; D003066:Coenzyme A Ligases / Q000235:genetics; D003373:Coumaric Acids / Q000378:metabolism*; D005285:Fermentation; D017353:Gene Deletion; D005947:Glucose / Q000378:metabolism*; D010649:Phenylalanine / Q000096:biosynthesis / Q000235:genetics; D011422:Propionates; D016958:Pseudomonas putida / Q000235:genetics / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
chemical_list: D002934:Cinnamates; D003373:Coumaric Acids; D011422:Propionates; C029010:cinnamic acid; D014443:Tyrosine; D010649:Phenylalanine; D003066:Coenzyme A Ligases; C400923:feruloyl-coenzyme A synthetase; C495469:p-coumaric acid; D005947:Glucose