Genetic manipulation of acid formation pathways by gene inactivation in Clostridium acetobutylicum ATCC 824

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 3 Author for correspondence: George N. Bennet. Te...

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Published in:Microbiology (Society for General Microbiology) 1996-08, Vol.142 (8), p.2079-2086
Main Authors: Green, Edward M, Boynton, Zhuang L, Harris, Latonia M, Rudolph, Frederick B, Papoutsakis, Eleftherios T, Bennett, George N
Format: Article
Language:English
Subjects:
Acetates - metabolism
Bacteriology
Biological and medical sciences
Biology of microorganisms of confirmed or potential industrial interest
Biotechnology
Butyrates - metabolism
Chromosomes, Bacterial
Clostridium - genetics
Clostridium - growth & development
Clostridium - metabolism
Clostridium acetobutylicum
Escherichia coli
Fermentation
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Genetics
Kinetics
Microbiology
Mission oriented research
Phosphate Acetyltransferase - biosynthesis
Phosphate Acetyltransferase - genetics
Phosphotransferases (Carboxyl Group Acceptor) - biosynthesis
Phosphotransferases (Carboxyl Group Acceptor) - genetics
Plasmids
Recombination, Genetic
Species Specificity
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Summary: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 3 Author 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. Keywords: Clostridium acetobutylicum , solvent biosynthesis, acid biosynthesis, metabolic engineering, gene integration
ISSN:1350-0872
1465-2080
DOI:10.1099/13500872-142-8-2079