Metabolic routing towards polyhydroxyalkanoic acid synthesis in recombinant Escherichia coli (fadR): inhibition of fatty acid beta-oxidation by acrylic acid.
Heterologous expression of the phaC1 gene from Pseudomonas aeruginosa, which encodes one of the polyhydroxyalkanoic acid synthases, in Escherichia coli impaired in fatty acid beta-oxidation results in polyhydroxyalkanoic acid accumulation when cells were cultivated on fatty acids. We evaluated the application of the fatty acid beta-oxidation inhibitor acrylic acid as a tool to channel intermediates of beta-oxidation to polyhydroxyalkanoic acid synthesis. Various E. coli strains affected in fatty acid metabolism and the wild-type strain harboring plasmid pBHR71 were analyzed with respect to polyhydroxyalkanoic acid accumulation in the presence of acrylic acid. The E. coli fadR mutant RS3097 revealed the strongest polyhydroxyalkanoic acid accumulation. The optimum inhibitory concentration of acrylic acid was 0.24 mg ml-1 and caused efficient channeling of intermediates of beta-oxidation to polyhydroxyalkanoic acid synthesis. Under these conditions and grown on decanoate E. coli RS3097 harboring plasmid pBHR71 revealed a polyhydroxyalkanoic acid accumulation contributing to about 60% of cellular dry weight.
Institut für Mikrobiologie, Westfälische Wilhelms-Universität, Münster, Germany.
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D000179:Acrylates / Q000494:pharmacology; D000217:Acyltransferases / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002264:Carboxylic Acids / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism*; D005285:Fermentation; D005798:Genes, Bacterial; D009154:Mutation; D010084:Oxidation-Reduction; D010957:Plasmids / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics; D011995:Recombination, Genetic; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*
D000179:Acrylates; D001426:Bacterial Proteins; D002264:Carboxylic Acids; C057119:FadR protein, Bacteria; D005227:Fatty Acids; D012097:Repressor Proteins; D000217:Acyltransferases; C067032:poly(3-hydroxyalkenoate)polymerase; C036658:acrylic acid