title: Improving endoglucanase activity by adding the carbohydrate-binding module from Corticium rolfsii.
abstract: The carbohydrate-binding module (CBM) is an important domain of most cellulases that plays a key role in the hydrolysis of cellulose. The neutral endoglucanase (EG1) gene was reconstructed. A redesigned endoglucanase, named EG2, was constructed with a CBM containing a linker from Corticium rolfsii (GenBank Accession No. D49448). The redesigned EG genes were expressed in Escherichia coli, and their characteristics are discussed. Results showed that the degradation of cellulose by EG2 was about double that by EG1. The specific activities of EG1 and EG2 were tested under optimal conditions, and EG2 had higher activity (169.1 ± 2.74 U/mg) toward CMC-Na than did EG1 (84.0 ± 1.98) in the process of cellulose degradation. The optimal pH and temperature, pH stability, and heat stability of EG1 and EG2 were similar. Results indicated that the CBM plays an essential role in the hydrolysis of cellulose. We can improve EG's catalytic power by adding the CBM from Corticium rolfsii.
authors.affiliation: College of Life and physical Sciences, Sichuan Agricultural University, Yaan 625014, China.
authors.forename: Zizhong
authors.initials: Z
authors.lastname: Tang
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.forename: Lijiao
authors.initials: L
authors.lastname: Chen
authors.forename: San
authors.initials: S
authors.lastname: Liu
authors.forename: Xueyi
authors.initials: X
authors.lastname: Han
authors.forename: Qi
authors.initials: Q
authors.lastname: Wu
mesh_terms: D001487:Basidiomycota / Q000201:enzymology* / Q000235:genetics; D002480:Cellulase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature
chemical_list: D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C055213:saccharide-binding proteins; D002482:Cellulose; D002480:Cellulase