Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli.
Acetyl-CoA carboxylase (ACC) catalyzes the first committed step of the fatty acid synthetic pathway. Although ACC has often been proposed to be a major rate-controlling enzyme of this pathway, no direct tests of this proposal in vivo have been reported. We have tested this proposal in Escherichia coli. The genes encoding the four subunits of E. coli ACC were cloned in a single plasmid under the control of a bacteriophage T7 promoter. Upon induction of gene expression, the four ACC subunits were overproduced in equimolar amounts. Overproduction of the proteins resulted in greatly increased ACC activity with a concomitant increase in the intracellular level of malonyl-CoA. The effects of ACC overexpression on the rate of fatty acid synthesis were examined in the presence of a thioesterase, which provided a metabolic sink for fatty acid overproduction. Under these conditions ACC overproduction resulted in a 6-fold increase in the rate of fatty acid synthesis.
Davis
M S
MS
Departments of Microbiology and Biochemistry, University of Illinois, Urbana, Illinois 61801, USA.
Solbiati
J
J
Cronan
J E
JE
D000103:Acetyl-CoA Carboxylase / Q000502:physiology*; D004926:Escherichia coli / Q000378:metabolism*; D005227:Fatty Acids / Q000096:biosynthesis*
D005227:Fatty Acids; D000103:Acetyl-CoA Carboxylase