Expression, Purification, Biochemical Characterization And Structural Modeling Of An Endo-Beta-1,4-Glucanase From Stachybotrys Chartarum In Pichia Pastoris

In this study, we analyzed the physicochemical characteristics and the structure of one glucanase and hoped it will be useful for future studies on the possible synergistic action of cellulases and it will provide guidance for the industrial utilization. A recombinant glucanase gene (r-ScEG12) from Stachybotrys chartarum was synthesized and expressed in Pichia pastoris, then was purified and characterized. The r-ScEG12 belongs to glycosyl hydrolase family 12 (GH12) by phylogenetic analysis. The Asp100, Glu117, and G1u201 residues were proposed to be present at the active site. The apparent molecular weight of r-ScEG12 was approximate 27 kDa, and the optimum temperature and pH was 40 degrees C and 5.0, respectively. The Km and Vmax for CMC were 26.08 g/L, and 1.88 mg L(-1)min(-1), respectively. The r-ScEG12 activity was inhibited 71.64 % and 50.97% by Cu2+ and Mn2+ respectively, while the r-ScEG12 activity was enhanced, by Na+ and CO+ mildly.