The hydrolysis and fermentation reaction were performed in the rotary flask shaker (Model: LRD-750) with
working volume 100ml in 250ml flasks. Based on the study purposes various amount of substrate with different
particle size were taken in the flask. All the composition and parameter assumed same in the total reaction mixture
due to continuous rotation. All experiments were run with different predetermined amount of enzyme in the
hydrolysis environment with different amount of substrate, different size of particle, different pH, and different
temperature to optimize the corresponding parameter. Samples were taken time to time and boiled with DNS
solution after centrifuging the sample to destroy the enzymes activity which confirms the reaction ceasing. Then the
sample was analysed for glucose in UV spectrophotometer according to the method by Miller [12]. When the
hydrolysis reaction reached at its equilibrium then the reaction mixture were separated by filtration method and
performing fermentation to produce bio ethanol using Saccharomyces cerevisiae (Baker's yeast) and detect the
ethanol concentration using K2Cr2O7 reagent and UV spectrophotometric method proposed by Adran and Prifysgol
[13].
3. Result and discussion
Though the reaction rate depends on different parameters, but in this study we assumed that the rate is affected
only by