Integrational plasmid technology has been used to disrupt metabolic pathways leading to acetate and butyrate formation in ATCC 824. Non-replicative plasmid constructs, containing either clostridial phosphotransacetylase () or butyrate kinase ) gene fragments, were integrated into homologous regions on the chromosome. Integration was assumed to occur by a Campbell-like mechanism, inactivating either or Inactivation of the gene reduced phosphotransacetylase and acetate kinase activity and significantly decreased acetate production. Inactivation of the gene reduced butyrate kinase activity, significantly decreased butyrate production and increased butanol production.


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