Abstract
In Bacillus subtilis, the germination-related lipase LipC is located in the spore coat, and mutant spores are defective in L-alanine-stimulated germination. To determine the physiological role of LipC, the recombinant LipC expressed in Escherichia coli was purified and characterized. The enzyme hydrolyzes p-nitrophenyl ester substrates with various acyl-chain lengths. Thin-layer chromatography and gas chromatography-mass spectrometry analysis indicated that LipC cleaves the fatty acids at the sn-1 and sn-2 positions of phospholipids as phospholipase B, and that the enzyme shows no selectivity for the polar head groups of lipid molecules. When the amounts of free fatty acids in dormant wild-type and lipC mutant (YCSKd) spores were measured, the amount of free fatty acids in the YCSKd spores was about 35% less than in the wild-type spores. These results suggest the possibility that Bacillus subtilis LipC plays an important role in the degradation of the outer spore membrane during sporulation.
