Pl see

medcraveonline.com/JMEN/JMEN-01-00003.pdf

Thank you very much sir

With decades of studies on cellulose bioconversion, cellulases have been playing an important role in producing fermentable sugars from lignocellulosic biomass. Usually, cellulases are mainly composed of three types of synergistic enzymes: endoglucanases (EC 3.2.1.4) that hydrolyze the exposed cellulose chains of the cellulose polymer, exoglucanases (cellobiohydrolases, EC 3.2.1.91) that act to release cellobiose from the reducing and nonreducing ends, and β-glucosidases (EC 3.2.1.21) that help to cleave the cellobiose and short-chain cello-oligosaccharide into glucose.

The soil sample suspensions can be inoculated on Czapek’s medium containing sugarcane bagasse pulp (in g/L: NaNO3, 2; MgSO4·7H2O, 0.5; NaCl, 0.5; FeSO4·7H2O, 0.01; KH2PO4, 1.0; yeast extract, 0.4; pulp, 5 (containing 80% water); and agar, 15.0; pH 5.0) and incubate at 28°C. Subsequently, single colonies can be picked using an inoculating needle and inoculated onto Mandels and Reese medium containing carboxymethyl cellulose sodium salt (CMC-Na; in g/L: KH2PO4, 2.0; (NH4)2SO4, 1.4; MgSO4·7H2O, 0.3; CaCl2, 0.3; yeast extract, 0.4; FeSO4·7H2O, 0.005; MnSO4, 0.0016; ZnCl2, 0.0017; CoCl2, 0.002; CMC-Na, 5.0; and agar, 15.0; pH 5.0). After incubation at 28°C for 48 h, all the plates can be stained with 1% (w/v) Congo-red solution for 15 min and discolored with 1 M NaCl for 15 min [19]. The degradation zones remain visible around the bacteria, showing that the strains could hydrolyze CMC.

The modified Mandels medium (also called basal medium) is also used for CMCase production by the isolates contained the following components (in g/L: KH2PO4, 1.5; Na2HPO4·7H2O, 2.5; (NH4)2SO4, 1.5; MgSO4·7H2O, 0.3; CaCl2, 0.1; FeSO4·7H2O, 0.005; MnSO4, 0.0016; ZnCl2, 0.0017; and CoCl2, 0.002; pH 7.0). The bacterial isolates can be  precultured overnight in general bacteria medium (in g/L: beef extract, 2; yeast extract, 2; sucrose, 6; and peptone, 5) at 28°C and 180 rpm. Subsequently, 2 mL of the culture can be  inoculated into 250 mL conical flask containing 50 mL of basal medium with 10 g/L of CMC-Na as the sole carbon source and incubate at 28°C and 180 rpm for 60 h.