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Production of polyhydroxyalkanoates by Escherichia coli mutants with defected mixed acid fermentation pathways



A series of Escherichia coli BW25113 mutants with reduced mixed acid fermentation were constructed. Genes ackA-pta , poxB , ldhA , adhE , and pflB encoding acetate kinase, phosphate acetyltransferase, pyruvate oxidase, d -lactate dehydrogenase, acetaldehyde dehydrogenase, and pyruvate formate-lyase, respectively, were deleted successively. When grown under microaerobic condition, the mutants reduced approximately 90% acetate excretion after the deletion of genes a ckA-pta and poxB . Production of lactate, ethanol, and formate was also significantly reduced after the deletion of genes ldhA , adhE , and pflB , respectively. The accumulation of biomass and poly(3-hydroxybutyrate) (PHB) were significantly enhanced after deleting the mixed acid fermentation. E. coli mutant BWapld with deletions of ackA-pta , poxB , ldhA , and adhE produced twice the cell dry weight (CDW) and 3.5 times of PHB compared with its wild-type under microaerobic conditions. E. coli mutant BWapl with deletions of ackA-pta , poxB , and ldhA also achieved nearly twice CDW and three times of PHB content in comparison to the wild-type during 48 h static cultivation. Production of poly(3-hydroxybutyrate- co -3-hydroxyvalerate) (P(3HB- co -3HV)) was observed in the mutants under static cultivation. E. coli mutant BWapld could produce approximately 50 wt.% P(3HB- co -3HV) consisting of 5 mol% of 3-hydroxyvalerate (3HV) under aerobic conditions, when the seed culture was inoculated at an appropriate time. When ackA-pta , poxB , ldhA , adhE , and pflB were deleted, E. coli mutant BWapldf accumulated over 70 wt.% P(3HB- co -3HV) consisting of 8 mol% 3HV under aerobic conditions.



Applied Microbiology and BiotechnologySpringer Journals

Published: Aug 1, 2010

DOI: 10.1007/s00253-010-2706-0

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