Green E M, Boynton Z L, Harris L M, Rudolph F B, Papoutsakis E T, Bennett G N
Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77005-1892, USA.
Microbiology (Reading). 1996 Aug;142 ( Pt 8):2079-86. doi: 10.1099/13500872-142-8-2079.
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.
整合质粒技术已被用于破坏丙酮丁醇梭菌ATCC 824中导致乙酸盐和丁酸盐形成的代谢途径。含有梭菌磷酸转乙酰酶（pta）或丁酸盐激酶（buk）基因片段的非复制性质粒构建体被整合到染色体上的同源区域。整合被认为是通过类似坎贝尔的机制发生的，使pta或buk失活。pta基因的失活降低了磷酸转乙酰酶和乙酸激酶的活性，并显著降低了乙酸盐的产生。buk基因的失活降低了丁酸盐激酶的活性，显著降低了丁酸盐的产生并增加了丁醇的产生。
