从昆虫病原真菌蜡蚧轮枝菌中分离、部分鉴定及克隆一种细胞外几丁质酶
Isolation, partial characterization, and cloning of an extracellular chitinase from the entomopathogenic fungus Verticillium lecanii.
作者信息
Yu G, Xie L Q, Li J T, Sun X H, Zhang H, Du Q, Li Q Y, Zhang S H, Pan H Y
机构信息
College of Plant Science, Jilin University, Changchun, China.
College of Resource and Environment, Jilin Agricultural University, Changchun, Jilin, China.
出版信息
Genet Mol Res. 2015 Mar 27;14(1):2275-89. doi: 10.4238/2015.March.27.13.
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.
昆虫病原真菌蜡蚧轮枝菌是一种著名的真菌植物病原体以及害虫的生物防治剂。分离出了一种属于糖基水解酶18家族的42 kDa几丁质酶，并对其进行了部分特性分析。使用苯基琼脂糖、DEAE琼脂糖和CM琼脂糖通过连续柱色谱法纯化几丁质酶。该酶在40°C和pH 4.6时表现出最高活性。在Mg(2+)存在下，酶活性被强烈激活。纯化后的酶对几种植物病原体，特别是串珠镰刀菌的孢子萌发具有抑制活性。通过聚合酶链反应扩增和测序解析了基因组DNA和cDNA序列。不同几丁质酶氨基酸的蛋白质建模和比较研究表明，几丁质酶在寄生真菌中是保守的。
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