Control of acetate production rate in Escherichia coli by regulating expression of single-copy pta using lacI(Q) in multicopy plasmid.
A tightly regulated gene expression system composed of a single-copy target gene under the control of a lac promoter derivative and lacI gene in a multicopy plasmid is proposed, and its ability to control the flux of a metabolic pathway is demonstrated. A model system to control the flux of acetyl-CoA to acetyl phosphate was constructed by integrating pta, a gene encoding phosphotransacetylase, under a tac promoter into the chromosome of E. coli with a pta-negative background and transforming a multicopy plasmid containing the lac iota(Q) gene into the strain. The production rate of acetate was shown to be tightly controlled when varying the concentration of the inducer (IPTG) in the model system.
Lee
Sun-Gu
SG
Department of Chemical and Biochemical Engineering, Pusan National University, Busan, 609-735, Korea. sungulee@pusan.ac.kr
Liao
James C
JC
D000085:Acetates / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D018628:Gene Dosage; D015964:Gene Expression Regulation, Bacterial; D007544:Isopropyl Thiogalactoside / Q000378:metabolism; D007763:Lac Operon; D056968:Lac Repressors; D010709:Phosphate Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*
D000085:Acetates; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D056968:Lac Repressors; C539064:LacI protein, E coli; D012097:Repressor Proteins; D007544:Isopropyl Thiogalactoside; D010709:Phosphate Acetyltransferase