PMID:9721301
| Citation |
Subrahmanyam, S and Cronan, JE Jr (1998) Overproduction of a functional fatty acid biosynthetic enzyme blocks fatty acid synthesis in Escherichia coli. J. Bacteriol. 180:4596-602 |
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| Abstract |
beta-Ketoacyl-acyl carrier protein (ACP) synthetase II (KAS II) is one of three Escherichia coli isozymes that catalyze the elongation of growing fatty acid chains by condensation of acyl-ACP with malonyl-ACP. Overexpression of this enzyme has been found to be extremely toxic to E. coli, much more so than overproduction of either of the other KAS isozymes, KAS I or KAS III. The immediate effect of KAS II overproduction is the cessation of phospholipid synthesis, and this inhibition is specifically due to the blockage of fatty acid synthesis. To determine the cause of this inhibition, we examined the intracellular pools of ACP, coenzyme A (CoA), and their acyl thioesters. Although no significant changes were detected in the acyl-ACP pools, the CoA pools were dramatically altered by KAS II overproduction. Malonyl-CoA increased to about 40% of the total cellular CoA pool upon KAS II overproduction from a steady-state level of around 0.5% in the absence of KAS II overproduction. This finding indicated that the conversion of malonyl-CoA to fatty acids had been blocked and could be explained if either the conversion of malonyl-CoA to malonyl-ACP and/or the elongation reactions of fatty acid synthesis had been blocked. Overproduction of malonyl-CoA:ACP transacylase, the enzyme catalyzing the conversion of malonyl-CoA to malonyl-ACP, partially relieved the toxicity of KAS II overproduction, consistent with a model in which high levels of KAS II blocks access of the other KAS isozymes to malonyl-CoA:ACP transacylase. |
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| Keywords |
3-Oxoacyl-(Acyl-Carrier-Protein) Synthase/biosynthesis; 3-Oxoacyl-(Acyl-Carrier-Protein) Synthase/genetics; 3-Oxoacyl-(Acyl-Carrier-Protein) Synthase/metabolism; Acyl Carrier Protein/metabolism; Acyl-Carrier Protein S-Malonyltransferase; Acyltransferases/biosynthesis; Acyltransferases/metabolism; Bacterial Proteins/genetics; Base Sequence; Coenzyme A/metabolism; DNA Primers; Escherichia coli/genetics; Escherichia coli Proteins; Esters; Fatty Acid Synthetase Complex, Type II; Fatty Acids/antagonists & inhibitors; Fatty Acids/biosynthesis; Lac Repressors; Operator Regions, Genetic; Promoter Regions, Genetic; Repressor Proteins/genetics |
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