title: Isolation and characterization of a novel endoglucanase from a Bursaphelenchus xylophilus metagenomic library.
abstract: A novel gene (designated as cen219) encoding endoglucanase was isolated from a Bursaphelenchus xylophilus metagenomic library by functional screening. Sequence analysis revealed that cen219 encoded a protein of 367 amino acids. SDS-PAGE analysis of purified endoglucanase suggested that Cen219 was a monomeric enzyme with a molecular mass of 40 kDa. The optimum temperature and pH for endoglucanase activity of Cen219 was separately 50 °C and 6.0. It was stable from 30 to 50 °C, and from pH 4.0 to 7.0. The activity was significantly enhanced by Mn(2+) and dramatically reduced by detergent SDS and metals Fe(3+), Cu(2+) or Hg(2+). The enzyme hydrolyzed a wide range of β-1, 3-, and β-1, 4-linked polysaccharides, with varying activities. Activities towards microcrystalline cellulose and filter paper were relatively high, while the highest activity was towards oat gum. The Km and Vmax of Cen219 towards CMC was 17.37 mg/ml and 333.33 U/mg, respectively. The findings have an insight into understanding the molecular basis of host-parasite interactions in B. xylophilus species. The properties also make Cen219 an interesting enzyme for biotechnological application.
authors.affiliation: Department of Life Science and Biotechnology, Nanyang Normal University, Nanyang, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Life Science and Biotechnology, Nanyang Normal University, Nanyang, China.
authors.forename: Yongxin
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Department of Life Science and Biotechnology, Nanyang Normal University, Nanyang, China.
authors.forename: Haoying
authors.initials: H
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Motor Bio-fuel technology, Henan Tianguan Group Co. Ltd., Nanyang, China.
authors.forename: Fengguang
authors.initials: F
authors.lastname: Du
authors.affiliation: Laboratory for Conservation and Utilization of Bio-Resources, and Key Laboratory for Microbial Resources of the Ministry of Education, Yunnan University, Kunming, China.
authors.forename: Ke-qin
authors.initials: KQ
authors.lastname: Zhang
authors.affiliation: Laboratory for Conservation and Utilization of Bio-Resources, and Key Laboratory for Microbial Resources of the Ministry of Education, Yunnan University, Kunming, China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Motor Bio-fuel technology, Henan Tianguan Group Co. Ltd., Nanyang, China.
authors.forename: Linfeng
authors.initials: L
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Motor Bio-fuel technology, Henan Tianguan Group Co. Ltd., Nanyang, China.
authors.forename: Man
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Life Science and Biotechnology, Nanyang Normal University, Nanyang, China.
authors.forename: Qiuhong
authors.initials: Q
authors.lastname: Niu
mesh_terms: D000818:Animals; D002480:Cellulase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015698:Genomic Library; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D054892:Metagenome; D009348:Nematoda / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny
chemical_list: D015801:Helminth Proteins; D002480:Cellulase