was performed in a water bath shaker at 27±2°C for 24 h. The
resultant supernatants following centrifugation (2500 g, 15 min)
were assayed for total reducing sugars by DNS method (www.glue.
umd.edu/~nsw/ench485/lab4a.htm). The sugars released were expressed
as equivalent of glucose.
The optimum temperature of saccharification was determined by
incubating the reaction mixture at different temperature ranging
from 25 to 60°C. The optimum pH was determined by adjusting the
pH of the reaction mixture from 3.5 to 6.5. The optimum substrate
concentration was determined by preparing substrate suspensions
1 to 6%.