FIG 3.

Hydrolytic activity of minicellulsomes. (A) The different strains were cultured in sPASC media (1% PASC) for 18 h at 30°C. The PASC degradation could be seen from the decrease in amounts of residual PASC in both three-enzyme and five-enzyme stains. The addition of LPMO and CDH in the five-enzyme strain enhanced cellulolytic activity of the cells. (B) The different strains were cultured in hydrolysis media (0.1% PASC) for 4 h at 30°C. The starting OD of the cultures was 10. For some of the cultures, 100 mM methyl glyoxal was added to inhibit glucose metabolism. In the absence of methyl glyoxal, any glucose produced by PASC hydrolysis was utilized by the cells resulting in low glucose concentrations. In the presence of methyl glyoxal, glucose utilization is inhibited and glucose produced by PASC hydrolysis is accumulated in the media. The five-enzyme strain had significantly higher cellulose hydrolysis activity, as indicated by the larger amounts of glucose released. Values represent the means of three samples. Error bars denote standard deviations. *, P < 0.05 (Student t test).