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...

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Published inJournal of pure & applied microbiology : an international research journal of microbiology Vol. 10; no. 1; p. 1
Main Authors You, Shuang-Hong, He, Yong-Mei, Zhu, Bo, Wang, Bo, Li, Da-Wei, Peng, Ri-He, Yao, Quan-Hong
Format Journal Article
LanguageEnglish
Published Oriental Scientific Publishing Company 01.03.2016
Subjects
Ascomycota
Gene expression
Genetic aspects
Observations
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Summary: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 Glu201 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.sup.-1][min.sup.-1], respectively. The r-ScEG12 activity was inhibited 71.64 % and 50.97% by [Cu.sup.2+] and [Mn.sup.2+] respectively, while the r-ScEG12 activity was enhanced by Na+ and [Ca.sup.2+] mildly. Key words: Endo-[beta]-1,4-glucanase; Biochemical characterization; Structural Modeling; Pichia pastoris; Stachybotrys chartarum.
ISSN:0973-7510