Raw reducing sugar quantification

The enzyme-enriched supernatant of P. pastoris expression was incubated with CMC solution and the release of reducing sugar by the hydrolytic process was quantified in the DNS assay. The supernatant was incubated with the cellulosic substrate for up to 20 minutes. Uninoculated YPD medium and EV-transfected inoculated medium were also detected to discard residual reducing sugars present in the reaction medium and in the basal activity of other P. pastoris hydrolases. TlicBgluc1 activity can enrich the reaction medium in glucose equivalents approximately twice as much as the control (2.13 mM glucose to 4.76 mM glucose).