Proteolytic Activation and Inactivation of Chitin Synthetase from Mucor rouxii

Abstract

Crude chitin synthetase preparations from the mycelial and yeast forms of M. rouxii behaved differently. The mycelial preparations, incubated at 28.degree. C, lost virtually all chitin synthetase activity in a few hours; by contrast, the activity of enzyme preparations from yeast cells increased several fold during similar incubations. These spontaneous changes were probably caused by endogenous protease(s). Seemingly, the chitin synthetase in yeast preparations was present mainly in a latent, zymogenic, form that was activated by proteases. In the mycelial preparations, chitin synthetase was present mainly in an active state and was rapidly degraded by endogenous proteolysis. Exogenous proteases accelerated activation and destruction of chitin synthetase; an acid protease from Rhizopus chinensis was the most effective activator. The activation of chitin synthetase was inhibited by a soluble protein in the cell-free extract. Treatment with the detergent Brij 36T stabilized the chitin synthetase of crude preparations against spontaneous changes. Stabilized preparations were rapidly activated by exogenous proteases. The different behavior of chitin synthetases in crude extracts of mycelium and yeast cells is consistent with, and perhaps partially responsible for, the differences in wall construction between mycelial and yeast forms of M. rouxii.