Incubation time (IT), enzyme dose (ED), and substrate concentration (SC) were optimized for enhanced hydrolysis of the biomass using a fungal consortium enzyme.
Incubation times ranging from 24 h to 96 h were tested to find out the optimal duration for the maximum release of RS (mg/g of substrate).
Enzyme concentrations ranging from 2.5 to 15 FPU/mg per gram of substrate were used to assess the effect of ED.
Due to economic constraint ED values above 15 FPU were not investigated.
In all hydrolytic reactions, enzyme supplementing refers to protein content per gram of the pretreated biomass.
The effects of SC ranging from 1% to 2% (w/v) were also examined.
A typical non-optimized hydrolysis reaction consisted of 1% SC supplemented with 10 mg fungal consortium enzyme added to 10 mL of sodium citrate buffer (pH 5.0).
This reaction occurred in a rotary shaker (150 rpm) at 30 C.