with 5 and 10% v/v of pre-inoculum. Pretreated sugar cane bagasse
(20 g/L) was used as a carbon source for cellulase production.
The nominal and working volumes were 10 and 7 L, respectively.
The temperature, agitation and pH were maintained at 30 ◦C,
200–350 rpm and 5.0, respectively. Air was sparged at 0.5–1 vvm
to provide dissolved oxygen concentrations within 20–40% of the
saturated level. Samples were withdrawn every 12 h for quantification
of FPase, Avicelase, CMCase and -glucosidase activities, as
described by Eveleigh et al. (2009), NREL (Adney and Baker, 1996)
and Ghose (1987). Total protein content was measured using the
Bio-Rad protein assay (Bio-Rad Laboratories), which is based on the
Bradford method. BSA was used as standard. All experiments were
performed in duplicate and the samples analyses were performed
in triplicate.