title: Molecular cloning, expression, purification and characterization of a novel cellulase gene (Bh-EGaseI) in the beetle Batocera horsfieldi.
abstract: Wood-feeding insects depend heavily on the secretion of a combination of cellulases, mainly endoglucanases and other glucanases such as exoglucanases and xylanases, to achieve efficient digestion of the cellulose of cellulosic materials. In this paper, we report a novel cellulose Bh-EGaseI belonging to the glycoside hydrolase family 45(gh45-1) obtained from the beetle Batocera horsfieldi. The Bh-EGaseI gene spans 714 bp and consists of three exons coding 237 amino acid residues. The cDNA encoding Bh-EGaseI was expressed as 25 KDa in baculovirus-infected Bombyx mori larvae. The expression products of Bh-EGaseI from larval hemolymph showed a specific enzymatic activity of approximately 1030.87 IU per mg. The enzyme was active over a wide range of pH and temperatures; optimal activity was observed at 40 °C and pH 4.0. The effects of ions on Bh-EGaseI activity were also studied, and results indicated that activity decreased to different extents upon addition of ions. Investigations on Bh-EGaseI facilitate their potential application in the production of bioenergy and biomaterials from cellulosic biomass in the future.
authors.affiliation: Jiangsu University of Science and Technology, Zhenjiang, Jiangsu 212018, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu 212018, China.
authors.forename: Hui-Zhen
authors.initials: HZ
authors.lastname: Mei
authors.affiliation: Jiangsu University of Science and Technology, Zhenjiang, Jiangsu 212018, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu 212018, China. Electronic address: xiadingguo@126.com.
authors.forename: Ding-Guo
authors.initials: DG
authors.lastname: Xia
authors.affiliation: Jiangsu University of Science and Technology, Zhenjiang, Jiangsu 212018, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu 212018, China.
authors.forename: Qiao-Ling
authors.initials: QL
authors.lastname: Zhao
authors.affiliation: Jiangsu University of Science and Technology, Zhenjiang, Jiangsu 212018, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu 212018, China.
authors.forename: Guo-Zheng
authors.initials: GZ
authors.lastname: Zhang
authors.affiliation: Jiangsu University of Science and Technology, Zhenjiang, Jiangsu 212018, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu 212018, China.
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: Qiu
authors.affiliation: Jiangsu University of Science and Technology, Zhenjiang, Jiangsu 212018, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu 212018, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Qian
authors.affiliation: Institute of Sericulture and System Biology, Southwest University, Chongqing 400716, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Lu
mesh_terms: D000818:Animals; D002480:Cellulase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D001517:Coleoptera / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D019476:Insect Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D019476:Insect Proteins; D011994:Recombinant Proteins; D002480:Cellulase
keywords: Baculovirus; Batocera horsfieldi; Cellulase; Endoglucanases; Silkworm