Characterization of the fatty acid synthetase system of Curtobacterium pusillum.
Curtobacterium pusillum contains 11-cyclohexylundecanoic acid as a major component of cellular fatty acids. A trace amount of 13-cyclohexyltridecanoic acid is also present. Fatty acids other than omega-cyclohexyl fatty acids present are 13-methyltetradecanoic, 12-methyltetradecanoic, n-pentadecanoic, 14-methylpentadecanoic, 13-methylpentadecanoic, n-hexadecanoic, 15-methylhexadecanoic, 14-methylhexadecanoic, and n-heptadecanoic acids. The fatty acid synthetase system of this bacterium was studied. Various 14C-labeled precursors were added to the growth medium and the incorporation of radioactivity into cellular fatty acids was analyzed. Sodium [14C]acetate and [14C]glucose were incorporated into almost all species of cellular fatty acids, the incorporation into 11-cyclohexylundecanoic acid being predominant. [14C]Isoleucine was incorporated into 12-methyltetradecanoic and 14-methylhexadecanoic acids: [14C]leucine into 13-methyltetradecanoic and 15-methylhexadecanoic acids; and [14C]valine into 14-methylpentadecanoic acid. [14C]-Shikimic acid was incorporated almost exclusively into omega-cyclohexyl fatty acids. The fatty acid synthetase activity of the crude enzyme preparation of C. pusillum was reconstituted on the addition of acyl carrier protein. This synthetase system required NADPH and preferentially utilized cyclohexanecarbonyl-CoA as a primer. The system was also able to use branched- and straight-chain acyl-CoAs with 4 to 6 carbon atoms effectively as primers but was unable to use acetyl-CoA. However, if acetyl acyl carrier protein was used as the priming substrate, the system produced straight-chain fatty acids. The results imply that the specificity of the initial acyl-CoA:acyl carrier protein acyltransferase dictates the structure of fatty acids synthesized and that the enzymes catalyzing the subsequent chain-elongation reactions do not have the same specificity restriction.
D000123:Acetyltransferases / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000378:metabolism; D051051:Acyl-Carrier Protein S-Acetyltransferase; D001419:Bacteria / Q000201:enzymology*; D002849:Chromatography, Gas; D004926:Escherichia coli / Q000201:enzymology; D064429:Fatty Acid Synthases / Q000032:analysis*; D005227:Fatty Acids / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity
D000214:Acyl Coenzyme A; D005227:Fatty Acids; D009249:NADP; D000123:Acetyltransferases; D051051:Acyl-Carrier Protein S-Acetyltransferase; D064429:Fatty Acid Synthases