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[家蚕几丁质酶的纯化与特性分析]

[Purification and characterization of a chitinase from Bombyx mori].

作者信息

Liu Mingyan, Zhang Hongbin, Hu Xueqin, Wei Qingli

机构信息

Department of Pharmaceutical Engineering, Hefei University of Technology, Hefei 230009, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2010 Mar;26(3):404-9.

Abstract

The importance of chitinases in the physiological and developmental processes of fungi and insects makes themselves and their inhibitors important targets for biological pesticides. A chitinase was isolated from Bombyx mori and purified to electrophoretic homogeneity by ammonium sulfate precipitation and Sephadex G-150 column chromatography. The molecular mass was estimated to be about 88 kDa by SDS-PAGE, while the K(m) was calculated to be 22.3 micromol/L. Moveover, the optimal reaction temperature was 45 degrees C, and the optimum pH was 6.0. The effect of metal ions and organic reagents on chitinase activity was investigated. The activity was enhanced by high concentration of Mn2+, while was strongly inhibited by Cu2+ and SDS. These results provide a basis for screening the chitinase-based biological pesticide.

摘要

几丁质酶在真菌和昆虫的生理及发育过程中的重要性,使得它们自身及其抑制剂成为生物农药的重要靶标。从家蚕中分离出一种几丁质酶,并通过硫酸铵沉淀和葡聚糖凝胶G - 150柱色谱法将其纯化至电泳纯。通过SDS - PAGE估计其分子量约为88 kDa,而计算得出的K(m)为22.3微摩尔/升。此外,最佳反应温度为45℃,最适pH为6.0。研究了金属离子和有机试剂对几丁质酶活性的影响。高浓度的Mn2 +可增强其活性,而Cu2 +和SDS则强烈抑制其活性。这些结果为筛选基于几丁质酶的生物农药提供了依据。

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