The solvent-tolerant Pseudomonas putida S12 as host for the production of cinnamic acid from glucose.
A Pseudomonas putida S12 strain was constructed that efficiently produced the fine chemical cinnamic acid from glucose or glycerol via the central metabolite phenylalanine. The gene encoding phenylalanine ammonia lyase from the yeast Rhodosporidium toruloides was introduced. Phenylalanine availability was the main bottleneck in cinnamic acid production, which could not be overcome by the overexpressing enzymes of the phenylalanine biosynthesis pathway. A successful approach in abolishing this limitation was the generation of a bank of random mutants and selection on the toxic phenylalanine anti-metabolite m-fluoro-phenylalanine. Following high-throughput screening, a mutant strain was obtained that, under optimised culture conditions, accumulated over 5 mM of cinnamic acid with a yield (Cmol%) of 6.7%.
Nijkamp
Karin
K
TNO Quality of Life, Business Unit Bioconversion and Processes for Food Industry, Apeldoorn, The Netherlands.
van Luijk
Nicole
N
de Bont
Jan A M
JA
Wery
Jan
J
D010723:3-Deoxy-7-Phosphoheptulonate Synthase / Q000378:metabolism; D002384:Catalysis; D002934:Cinnamates / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D004361:Drug Tolerance; D005285:Fermentation; D005818:Genetic Engineering; D005822:Genetic Vectors; D005947:Glucose / Q000378:metabolism*; D009154:Mutation; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000378:metabolism; D010650:Phenylalanine Ammonia-Lyase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011302:Prephenate Dehydratase / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D012997:Solvents / Q000494:pharmacology
D002934:Cinnamates; D012997:Solvents; C029010:cinnamic acid; C027218:3-fluorophenylalanine; D010649:Phenylalanine; D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D011302:Prephenate Dehydratase; D010650:Phenylalanine Ammonia-Lyase; D005947:Glucose