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Department of Biochemistry and Cell Biology, Institute of Biosciences and Bioengineering, Rice University, MS 140, 6100 Main Street, Houston, TX 77005-1892, USA
Department of Chemical Engineering, Northwestern University, Evanston, IL 60208-3120, USA
3Author for correspondence: George N. Bennet. Tel: +1 713 527 4920. Fax: +1 713 285 5154. e-mail: gbennett@bioc.rice.edu
ABSTRACT
Integrational plasmid technology has been used to disrupt metabolic pathways leading to acetate and butyrate formation in Clostridium acetobutylicum ATCC 824. Non-replicative plasmid constructs, containing either clostridial phosphotransacetylase (pta) or butyrate kinase (buk) gene fragments, were integrated into homologous regions on the chromosome. Integration was assumed to occur by a Campbell-like mechanism, inactivating either pta or buk. Inactivation of the pta gene reduced phosphotransacetylase and acetate kinase activity and significantly decreased acetate production. Inactivation of the buk gene reduced butyrate kinase activity, significantly decreased butyrate production and increased butanol production.
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