Solubilization and partial purification of yeast chitin synthetase. Confirmation of the zymogenic nature of the enzyme.

摘要: Chitin synthetase was solubilized with digitonin from a particulate yeast fraction. The enzyme, which did not sediment at 200,000 X g and emerged after the void volume in Sepharose 6B column, active only treatment protease. This confirms that chitin exists plasma membrane as zymogen initiation of septum occurs by localized activation enzyme. By differential extraction sodium cholate digitonin, followed chromatography on 6B, 20-fold purification enzyme achieved respect to crude particles. purified showed requirement for phospholipid; phosphatidylserine lysophosphatidylserine were best activators. Unsaturated fatty acids strongly inhibited activity, whereas their saturated counterparts inert. catalyzed formation insoluble absence added primer. synthetic polysaccharide examined electron microscopy found consist lozenge-shaped particles about 60 nm long 10 wide.