首页> 外文期刊>Journal of Microbiology, Biotechnology and Food Sciences >ZYMOGRAPHIC IDENTIFICATION AND BIOCHEMICAL CHARACTERIZATION OF CHITINASE AGAINST PHYTOFUNGAL PATHOGENS
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ZYMOGRAPHIC IDENTIFICATION AND BIOCHEMICAL CHARACTERIZATION OF CHITINASE AGAINST PHYTOFUNGAL PATHOGENS

机译:几丁质酶对植物真菌病原菌的形态学鉴定和生物化学表征

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An endospore forming Gram positive bacterium (MBCU4) was isolated from a vermicompost amended soil, and confirmed as Bacillus subtilis through the 16S rRNA sequence analysis. An extracellular chitinase was detected from this strain of B. subtilis under specific environmental condition. An attempt was made to purify the enzyme by ammonium sulfate precipitation followed by DEAE sepharose CL-6B column chromatography. The purified enzyme was demonstrated as a single band, having the molecular weight 31kDa on SDS PAGE analysis and its activity in the gel was determined by clear zone on zymogram. Further characterization of the isolated enzymes has showed that this enzyme is most active at pH 6.0 and at the optimized temperature of 50 0C. The purified chitinase exhibited high degree of antifungal activity particularly by degrading their cell wall components of plant pathogens Macrophomina phaseolina (69.0%) and Rhizoctonia solani (52.0%). It infers that the chitinase produced by B. subtilis could play an important role for biopesticidal activity.
机译:从a堆改良土壤中分离出形成内生孢子的革兰氏阳性细菌(MBCU4),并通过16S rRNA序列分析确认为枯草芽孢杆菌。在特定的环境条件下,从该枯草芽孢杆菌菌株中检​​测到细胞外几丁质酶。尝试通过硫酸铵沉淀,然后用DEAE琼脂糖CL-6B柱色谱法纯化酶。纯化的酶显示为单条带,在SDS PAGE分析中分子量为31kDa,其在凝胶中的活性通过酶谱图上的透明区确定。分离的酶的进一步表征表明,该酶在pH 6.0和最适温度50 0C下最具活性。纯化的几丁质酶表现出高度的抗真菌活性,尤其是通过降解植物病原体菜豆(Macrophomina phaseolina)(69.0%)和茄状根瘤菌(Rhizoctonia solani)(52.0%)的细胞壁成分。推测枯草芽孢杆菌产生的几丁质酶可能对杀虫活性起重要作用。

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