Genetic manipulation of acid formation pathways by gene inactivation in Clostridium acetobutylicum ATCC 824.
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.
Green
E M
EM
Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77005-1892, USA.
Boynton
Z L
ZL
Harris
L M
LM
Rudolph
F B
FB
Papoutsakis
E T
ET
Bennett
G N
GN
D000085:Acetates / Q000378:metabolism*; D002087:Butyrates / Q000378:metabolism*; D002876:Chromosomes, Bacterial; D003013:Clostridium / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004926:Escherichia coli; D005285:Fermentation; D005798:Genes, Bacterial; D007700:Kinetics; D010709:Phosphate Acetyltransferase / Q000096:biosynthesis* / Q000235:genetics; D017851:Phosphotransferases (Carboxyl Group Acceptor) / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids; D011995:Recombination, Genetic; D013045:Species Specificity
D000085:Acetates; D002087:Butyrates; D010709:Phosphate Acetyltransferase; D017851:Phosphotransferases (Carboxyl Group Acceptor); C019285:butyrate kinase