SUMMARY: 1,3-β-Glucan synthase activity has been detected in a membrane fraction extracted from the mycelium of the filamentous fungus The enzyme was solubilized by CHAPS and stabilized by filtration on a Bio-gel P30 column. Highest activity was obtained in the early exponential phase of growth. Four factors - GTP, NaF, sucrose and EDTA - added during the extraction procedure, were essential for optimal 1,3-β-glucan synthase activity. The soluble enzyme preparation was photolabelled with 5-azido-[P]UDP-glucose and 5-IASA-UDP-glucose which bind covalently to the enzyme after UV irradiation. These UDP-glucose substrate analogues were competitive inhibitors of the enzyme with a of 1.42 mM and 0.3 mM for 5-azido-UDP-glucose and 5-ASA-UDP-glucose, respectively for UDP-glucose = 1.9 mM). Potential UDP-glucose-binding polypeptides were identified with molecular masses of 31, 50 and 115 kDa.


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