title: cDNA cloning, expression, and enzymatic activity of a novel endogenous cellulase from the beetle Batocera horsfieldi.
abstract: In this study, we report a novel cellulase [β-1,4-endoglucanase (EGase), EC 3.2.1.4] cDNA (Bh-EGase II) belonging to the glycoside hydrolase family (GHF) 45 from the beetle Batocera horsfieldi. The Bh-EGase II gene spans 720bp and consists of a single exon coding for 239 amino acid residues. Bh-EGase II showed 93.72% protein sequence identity to Ag-EGase II from the beetle Apriona germari. The GHF 45 catalytic site is conserved in Bh-EGase II. Bh-EGase II has three putative N-glycosylation sites at 56-58 (N-K-S), 99-101 (N-S-T), and 237-239 (N-Y-S), respectively. The cDNA encoding Bh-EGase II was expressed in baculovirus-infected insect BmN cells and Bombyx mori larvae. Recombinant Bh-EGase II from BmN cells and larval hemolymph had an enzymatic activity of approximately 928U/mg. The enzymatic catalysis of recombinant Bh-EGase II showed the highest activity at 50°C and pH6.0.
authors.affiliation: Institute of Sericulture and System Biology, Southwest University, Chongqing, China.
authors.forename: Dingguo
authors.initials: D
authors.lastname: Xia
authors.forename: Yadong
authors.initials: Y
authors.lastname: Wei
authors.forename: Guozheng
authors.initials: G
authors.lastname: Zhang
authors.forename: Qiaoling
authors.initials: Q
authors.lastname: Zhao
authors.forename: Yeshun
authors.initials: Y
authors.lastname: Zhang
authors.forename: Zhonghuai
authors.initials: Z
authors.lastname: Xiang
authors.forename: Cheng
authors.initials: C
authors.lastname: Lu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000201:enzymology / Q000235:genetics; D020134:Catalytic Domain / Q000235:genetics; D002480:Cellulase / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D001517:Coleoptera / Q000201:enzymology* / Q000235:genetics*; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D017344:Genes, Insect; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D058974:Phylogeography; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D019476:Insect Proteins; D011994:Recombinant Proteins; D002480:Cellulase