Biosynthetic control of the natural abundance of carbon 13 at specific positions within fatty acids in Escherichia coli. Evidence regarding the coupling of fatty acid and phospholipid synthesis.

Stable carbon isotope ratios (13C/12C) at natural abundance levels have been determined for individual carbon atoms in each of the major phospholipid fatty acids of Escherichia coli grown on glucose as the sole carbon source. Two models were constructed for the isotope effects and carbon flow pathways which must be responsible for the observed isotopic fractionations. Both models incorporate a branch in the carbon flow at which fatty acyl-acyl carrier protein (acyl-ACP) is utilized either for complex lipid synthesis or for elongation by fatty acid synthetase. Depletion of carbon 13 in the carboxyl groups of myristic and palmitoleic acids (relative to carbonyl groups in precursor acyl-ACP's) was observed to occur at this branching site. Only one of the models was consistent both with this observation and with the observation (Silbert, D. F., Ruch, F., and Vagelos, P. R. (1968) J. Bacteriol. 95, 1658-1665) that exogenous fatty acids are incorporated into phospholipids but are not elongated. The successful model has free fatty acid as the intermediate product coupling fatty acid biosynthesis to phospholipid synthesis. Essential to this pathway are those reactions catalyzed by thioesterases I and II as well as acyl-ACP synthetase, enzymes whose roles have previously been unknown in vivo.