Cellulase production by bacillus spp and aspergillus niger using coir waste and saw dust and partial purification

In this study four bacterial colonies (Bacillus spp, two Pseudomonas spp and Proteus spp.) and two fungal spp (Aspergillus niger and Aspergillus fumigatus) were isolated by spread plate technique and were checked for cellulase production (zone formation) with the help of congo red and NaCl/NaOH. The production of cellulase by bacteria (Bacillus spp , Pseudomonas spp and Proteus spp) and fungi (A.niger and A. fumigatus) were also confirmed by different assay (Gel punch assay, Dinitrosalicylic acid (DNS) and Carboxy Methyl Cellulose assay (CMC) ). In DNS assay method high level of cellulase production was achieved by Bacillus spp (0.09 µl/ml and 0.08 µl/ml) for coir waste and saw dust as substrate and among fungi A. niger showed higher cellulase activity (0.1 µl/ml and 0.1 µl/ml) for coir waste and saw dust as substrate. In CMC assay high level of enzyme production was achieved by Bacillus spp (0.1 µl/ml and 0.09 µl/ml) for coir waste and saw dust and among fungi A.niger showed high level of cellulase activity (0.1µl/ml and 0.1µl/ml) .Regarding pH, Temperature, carbon source cellulase activity was high when pH was 6 for bacteria and 7 for fungi and for temp the optimum cellulase production was at 37ºC and 28ºC for bacteria and fungi. Among different carbon source cellulose showed higher yield in production for both bacteria and fungi. The crude enzyme after centrifugation of production media were used to determine the molecular weight of the enzyme produced which lied between 6 to 20 KDa for Bacillus for both substrate and that of A. niger was found to lie between 20 and 29 KDa for both the substrate by SDS-PAGE.