Production and Optimization of cellulase enzyme under Submerged and Solid State Fermentation from Trichoderma Isolates

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Addis Ababa University


Pure cellulose and agricultural wastes were used as sole carbon sources for the production of cellulase enzymes by Trichoderma isolates in both SmF and SSF, respectively. These isolates (seven) were obtained from Mycology laboratory, Faculty of Life science, Addis Ababa University. Carboxymethyl cellulose (CMC) and Congo red were used to screen four isolates for their ability to produce cellulase enzymes. Cellulase production was assayed by measuring the amount of glucose released in μmol/ml/min by using the DNS assay method at 540nm. To maximize the cellulase production, the critical parameters like carbon source, nitrogen source, cellulose concentration, temperature and pH were optimized under SmF. The highest cellulase enzyme activity was observed after 12 days of incubation on media containing, yeast extract (1%), cellulose concentration (1%) and pH (5.5) from all isolates under SmF whereas under SSF maximum cellulase enzyme was recorded between 5-11 days incubation. WB, RB and WS were found to suitable for high cellulase production whereas CS, CP and BB relatively showed the least cellulase production in SSF. Trichoderma isolates AUT1 produce the highest CMCase on WS (5.68U/g), AUT5 on rice bran (8.15U/g), AUT2 and AUT4 on wheat bran, their enzymatic activities were 4.92U/g and 7.01U/g, respectively. Enzyme synthesis was repressed in the presence of glucose and fructose while it was induced in the presence of maltose and lactose in SmF. The maximum amount of cellulase was observed between 55% to 65% moisture contents. It is evident from the present study, solid state fermentation was better than liquid state fermentation for the production of cellulase by Trichoderma isolates using agricultural wastes. The cellulase enzymes produced by Trichoderma isolates were active at temperature ranges of 40-60oC and pH values 4.5-6.5. Key words:- Agricultural wastes, Carboxymethyl cellulase, Cellulase activity, DNS methods and Congo red



Agricultural wastes, Carboxymethyl cellulase, Cellulase activity, DNS methods and Congo red