I need help with an extremely frustrating problem. I have separated
S. cerevisiae membrane extracts into the individual yeast phospholipids by
2-D TLC and now want to cleave them with bacterial PLC's to yield
diacylglycerol. The problem is that (unlike mammalian phospholipids) the
purified phospholipids appear to be resistant to hydrolysis by commercially
available PLCs from Bacillus cereus and Bacillus thuringiensis.
Has anybody had any similar experiences and overcome them ? Does
anybody have any information about yeast phospholipid head group
modifications which might lead to resistance to bacterial PLC hydrolysis ?
I hope someone can help.