Optimization of the solvent-tolerant Pseudomonas putida S12 as host for the production of p-coumarate from glucose.
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.
Nijkamp
Karin
K
Business Unit, Food and Biotechnology Innovations, TNO Quality of Life, P.O. Box 342, 7300 AH, Apeldoorn, The Netherlands. karin.nijkamp@tno.nl
Westerhof
R G Maaike
RG
Ballerstedt
Hendrik
H
de Bont
Jan A M
JA
Wery
Jan
J
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
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