Studies on cyclopropane fatty acid synthesis. Effect of carbon source and oxygen tension on cyclopropane fatty acid synthetase activity in Pseudomonas denitrificans.

The cyclopropane fatty acid, methylene hexadecanoic acid, constituted from 1% to upwards of 30% of the total lipid fatty acids of the bacterium, Pseudomonas denitrificans. The amount of this component varied along with the levels of the enzyme, cyclopropane synthetase (unsaturated-phospholipid methyltransferase, EC 2.1.1.16). When P. denitrificans was grown on succinate in a culture medium saturated with oxygen, cyclopropane synthetase remained repressed while cell densities were low. As cell densities increased, the enzyme was induced and the activity rose to a maximum over a period of 4-6 h. Cyclopropane synthetase could also be induced by rapidly limiting the oxygen supply to cells growing in conditions where oxygen was in excess. This phenomenon was independent of the phase of growth and could be prevented by addition of chloramphenicol to the medium. Growth on glucose was also shown to repress the synthesis of cyclopropane synthetase under similar conditions. However, once maximum levels of cyclopropane synthetase were reached, they remained constant for at least the following 15 h irrespective of the source of carbon in the medium. Methylene hexadecanoic acid accumulated in a linear manner throughout this period until a maximum level was achieved, the rate of accumulation being related to the activity of cyclopropane synthetase detected in vitro. The rate of conversion of total fatty acid to methylene hexadecanoic acid was approximately 1.3-1.5% per h, the methylene hexadecanoic acid being metabolically stable - the relative percentage of methylene hexadecanoic acid to total fatty acid in repressed cells, falling linearly with increase in cell number. Repression of enzyme synthesis was further investigated by growing cells on various sources of carbon other than glucose. The results indicated that succinate was unique amongst tricarboxylic acid cycle intermediates in depressing cyclopropane synthetase under limited oxygen conditions.