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).