Preparation of Cellulase Enzyme and Bio-ethanol Production from Agricultural Waste Product

Screening for cellulase-producing fungi was routinely done on cellulose yeast extract agar plates with 1% Congo red after 4 to 7 day old culture. After 20-30 minutes it could be seen a clear zone and then a little flooded with Gram's iodine to get a sharp and distinct clear zone around the cellulase producing fungal colonies. Grams iodine is the best plate assay method for determining cellulase activity and gives the best results with prominent and distinct clear zones within 3-5 minutes. Total (19) fungal isolates were screened from two selected areas such as (12) isolates from near the heap of straw soil, Mawbi Township and (7) isolates from the litter soil, Botanical garden of Yangon University, Kamayut Township. In this study, cellulolytic fungi belonged to Aspergillus sp., Penicillium sp., Fusarium sp., Spicaria sp., Rhizoctonia sp., Trichoderma sp. and Paecilomyces sp.. The effect of experimental factors is important for the production of reducing sugar by enzyme activity. In the present study, the effect of various fermentation periods (1, 2, 3, 4, 5, 6, 7 and 8 days), the effect of different temperature (20, 30, 40 and 50° C) and the effect of different pH (pH - 3, 4, 5, 6, 7 and 8) on the appearance of reducing sugar by the crude cellulase enzymes were studied and which were extracted from Aspergillus, penicillium and Fusarium, Spicaria sp. In this study, optimal fermentation period was 3 days in Aspergillus and Penicillium sp. and 4 days in Fusarium and Spicaria sp., optimal temperature 30° C of pH 6 in Aspergillus, Penicillium, Fusarium and Spicaria sp., respectively. The optimal experimental condition was carried out by crude cellulase enzymes using filter paper cellulose, rice straw and water hyacinth, and end products were analyzed by TLC method.