title: Isolation, partial characterization, and cloning of an extracellular chitinase from the entomopathogenic fungus Verticillium lecanii.
abstract: The entomopathogenic fungus Verticillium lecanii is a well-known biocontrol agent of fungal phytopathogens, as well as insect pests. A 42-kDa chitinase belonging to family 18 of the glycosyl hydrolases was isolated and partially characterized. Chitinase was purified using successive column chromatography on phenyl-sepharose, DEAE-sepharose, and CM-sepharose. The enzyme showed the highest activity at 40°C and pH 4.6. Enzyme activity was strongly activated in the presence of Mg(2+). The purified enzyme showed inhibitory activity of spore germination against several plant pathogens, particularly Fusarium moniliforme. The genomic DNA and cDNA sequences were resolved by polymerase chain reaction amplification and sequencing. Protein modeling and comparative investigation of different chitinase amino acids showed that chitinases are conserved in parasitic fungi.
authors: College of Plant Science, Jilin University, Changchun, China.
authors: G
authors: G
authors: Yu
authors: College of Plant Science, Jilin University, Changchun, China.
authors: L Q
authors: LQ
authors: Xie
authors: College of Plant Science, Jilin University, Changchun, China.
authors: J T
authors: JT
authors: Li
authors: College of Plant Science, Jilin University, Changchun, China.
authors: X H
authors: XH
authors: Sun
authors: College of Resource and Environment, Jilin Agricultural University, Changchun, Jilin, China.
authors: H
authors: H
authors: Zhang
authors: Jilin Academy of Agricultural Science, Changchun, China.
authors: Q
authors: Q
authors: Du
authors: Jilin Academy of Agricultural Science, Changchun, China.
authors: Q Y
authors: QY
authors: Li
authors: College of Plant Science, Jilin University, Changchun, China.
authors: S H
authors: SH
authors: Zhang
authors: College of Plant Science, Jilin University, Changchun, China panhongyu@jlu.edu.cn.
authors: H Y
authors: HY
authors: Pan
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D002688:Chitinases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002845:Chromatography / Q000295:instrumentation / Q000379:methods; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D005670:Fusarium / Q000187:drug effects / Q000254:growth & development; D006863:Hydrogen-Ion Concentration; D007313:Insecta / Q000382:microbiology*; D007700:Kinetics; D008969:Molecular Sequence Data; D012685:Sepharose; D017422:Sequence Analysis, DNA; D013172:Spores, Fungal / Q000187:drug effects / Q000254:growth & development; D013696:Temperature; D020172:Verticillium / Q000201:enzymology / Q000235:genetics*
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins; D012685:Sepharose; D002688:Chitinases