title: Improvement of the catalytic performance of a Bispora antennata cellulase by replacing the N-terminal semi-barrel structure.
abstract: The aim of this work was to study the contribution of the N-terminal structure to cellulase catalytic performance. A wild-type cellulase (BaCel5) of glycosyl hydrolase (GH) family 5 from Bispora antennata and two hybrid enzymes (BaCel5(127) and BaCel5(167)) with replacement of the N-terminal (βα)3 (127 residues) or (βα)4 (167 residues)-barrel with the corresponding sequences of TeEgl5A from Talaromyces emersonii were produced in Pichia pastoris and biochemically characterized. BaCel5 exhibited optimal activity at pH 5.0 and 50°C but had low catalytic efficiency (25.4±0.8mLs(-1)mg(-1)). In contrast, BaCel5(127) and BaCel5(167) showed similar enzymatic properties but improved catalytic performance. When using CMC-Na, barley β-glucan, lichenan, and cellooligosaccharides as substrates, BaCel5(127) and BaCel5(167) had increased specific activities and catalytic efficiencies by ∼1.8-6.7-fold and ∼1.0-4.7-fold, respectively. The catalytic efficiency of BaCel5(167) was even higher than that of parental proteins. The underlying mechanism was analyzed by molecular docking and molecular dynamic simulation.
authors.affiliation: College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, People's Republic of China; Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Fei
authors.initials: F
authors.lastname: Zheng
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Huoqing
authors.initials: H
authors.lastname: Huang
authors.affiliation: College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, People's Republic of China; Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Tu
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Kun
authors.initials: K
authors.lastname: Meng
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Su
authors.affiliation: College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, People's Republic of China.
authors.forename: Xiangming
authors.initials: X
authors.lastname: Xie
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China. Electronic address: luohuiying@caas.cn.
authors.forename: Huiying
authors.initials: H
authors.lastname: Luo
mesh_terms: D001203:Ascomycota / Q000201:enzymology*; D002384:Catalysis; D002480:Cellulase / Q000737:chemistry* / Q000378:metabolism; D003001:Cloning, Molecular; D005936:Glucans / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D062105:Molecular Docking Simulation; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D010843:Pichia / Q000378:metabolism; D000072417:Protein Domains; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity; D013696:Temperature; D047071:beta-Glucans / Q000737:chemistry / Q000378:metabolism
chemical_list: D005936:Glucans; D009844:Oligosaccharides; D047071:beta-Glucans; D002480:Cellulase; C015968:lichenin
keywords: Catalytic efficiency; Cellulase; Glycosyl hydrolase (GH) family 5; Hybrid enzyme; N-terminal (βα)-replacement