Abstract
Laboratory bench scaling was done and an average of 1.85 fold increase by Response Surface Methodology (RSM) optimization was obtained. It was found that the predicted value (4.96 IU/ml) obtained by RSM is in close accordance with observed activity 5.14 IU/ml. Endoglucanases are mainly induced by CMC while Wheat bran (natural substrate) exoglucanase is more active when induced by avicel and cellulose. Addition of substrate beyond a level caused inhibition of cellulase production. The molecular weight of protein as determined by SDS-PAGE is very similar to molecular weight of cellulase of Trichoderma viride (T. viride) cellulase and Trichoderma reesei (T. reesei) endoglucanase. T. reesei β-glucosidase has high enzymatic activity on CMC substrate when compared with T. viride β-glucosidase. Secondary structure analysed by using Circular Dichroism confirmed that composition of celluase system is very similar to other analysed species. The cellulase was found to be active in pH range of 4.8-5.5; while temperature range varied from 50°C to 70°C. Although the enzymatic activity produced by mutants were lesser than the parent, but in one case mutants of Trichoderma reesei’s BGL has shown higher activity on cellulose.
Highlights
Microbial cellulase have wide spread application in textile, pulp, brewing, food and agriculture industry
Isolation and identification of Actinomycetes Novel isolate from the soil sample collected from high altitude of Leh, Jammu & Kashmir, India was identified as actinomycetes by Gram staining and Methylene blue tests
Screening of the Trichoderma and Actinomycetes strains for cellulase production All the organisms under study which were incubated with congo red, produced cellulases as observed due to Coefficient t-value
Summary
Microbial cellulase have wide spread application in textile, pulp, brewing, food and agriculture industry. Because of its commercial importance and complexity of structure, research of many labs and industries are revolving around cellulase producing microbes, its production and its characterization. It has two domains, cellulose binding module (CBM) and a catalytic domain (CD). Cellulase family contain three groups of enzymes, endo(1,4)-β-D-glucanase (EG), exo-(1,4)-β-D-glucanase (CBH) and β-glucosidase (BGL). EG breaks o-glycosidic bonds which are present internally but randomly. CBH attacks cellulose chains at the ends, to release β-cellulobiose.
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