title: Purification and characterization of recombinant endoglucanase of Trichoderma reesei expressed in Saccharomyces cerevisiae with higher glycosylation and stability.
abstract: Cel5A (endoglucanase II) of Trichoderma reesei was expressed in Saccharomyces cerevisiae then purified. Two components (C1 and C2) of recombinant Cel5A with different glycosylation were obtained. Purified C1 had a larger molecular mass (57 kDa) than that of the native Cel5A produced by T. reesei (48 kDa) due to the different extents of asparagines-linked glycosylation. There was no significant difference in enzymatic activity between the C1 and the native Cel5A from T. reesei. C1 treated with Endoglycosidase H had a molecular mass of 54 kDa and retained about 88% of its original activity. Unpurified C2 was larger form of hyperglycosylation proteins. Its molecular mass was larger than 85 kDa till up to 200 kDa. It still retained activity regardless of its magnitude molecular mass. With increased glycosylation extent of the enzyme components (C2 >C1 >native Cel5A), the pH range of activity become wider, and thermal stability become higher.
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, PR China.
authors.forename: Yuqi
authors.initials: Y
authors.lastname: Qin
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Wei
authors.forename: Xiangmei
authors.initials: X
authors.lastname: Liu
authors.forename: Tianhong
authors.initials: T
authors.lastname: Wang
authors.forename: Yinbo
authors.initials: Y
authors.lastname: Qu
mesh_terms: D002480:Cellulase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D003001:Cloning, Molecular; D004795:Enzyme Stability; D006031:Glycosylation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; C056373:endoglucanase 2; D002480:Cellulase