Effect of tunicamycin on N-acetyl-beta-D-glucosaminidase produced by Trichoderma harzianum

The effect of tunicamycin, an inhibitor of protein N-glycosylation, was studied in non-growing mycelium of Trichoderma harzianum induced to secrete N-acetyl-beta-D-glucosaminidase by the addition of N-acetylglucosamine. Tunicamycin (30 microg ml(-1)) had no significant effect on growth of the fungus, or on the total protein secreted or specific activity of N-acetyl-beta-D-glucosaminidase. However, in the presence of the inhibitor an underglycosylated form of the enzyme was produced. The apparent molecular masses for this and the native enzyme were 110 and 124 kDa, respectively. Both forms of the enzyme showed the same optimum pH and temperature, but the underglycosylated form was more sensitive to inactivation by both high temperature (60 degrees C) and the proteolytic enzyme trypsin.