A particulate chitin synthase from Aspergillus flavus Link: the properties, location, and levels of activity in mycelium and regenerating protoplast preparations

Abstract

Chitin synthase (ED 2.4.1.16) has been characterized in Aspergillus flavus. A K(m) value of 2.5 m(M) was obtained for the substrate UDPGlcNAc. The enzyme had a requirement for GlcNAc, and Mg2+ and activity was increased in the presence of soluble chitodextrins F1 and F2. The optimum activity was obtained using Tris--HCl buffer, pH 7.5, with a secondary peak at pH 6.2 and an incubation temperature of 29.5 degrees C. Distribution patterns of chitin synthase in protoplasts and mycelial material were very similar. The highest specific activity was found in a 200 000 X g fraction. Enzyme levels in growing mycelium increased during the exponential growth phase after which they declined. Activity also increased during the early stages of regeneration of both conidial and mycelial protoplasts, despite an initial lack in net protein synthesis. Chitin synthase levels were also dependent upon the carbon source available during regeneration.