红豆中几丁质酶的纯化与表征  

Purification and characterization of a chitinase from the adzuki bean

作　　者：刘心义[1] 徐二华[1] 陈继伟[1] 刘元刚[1] 汪少芸[1] 叶秀云[1] 

机构地区：[1]福州大学生物工程研究所,福建福州350002

出　　处：《福州大学学报（自然科学版）》2007年第4期625-630,共6页Journal of Fuzhou University(Natural Science Edition)

摘　　要：利用硫酸铵沉降、离子交换色谱CM-Sephadex C-50、凝胶过滤色谱Sephadex G-75以及高效液相色谱(HPLC)Poros HS-20,从红豆种子中分离纯化出一种具有抗真菌活性的几丁质酶.SDS-PAGE显示还原与非还原状态下该蛋白分子质量分别为27.7和22.7 kDa,这表明该几丁质酶分子内含有二硫键.该酶比活为14.57 U/mg,最适反应pH为5.4,最适反应温度为50℃.它对棉花枯萎病菌、甜瓜枯萎病菌与瓜果腐霉病菌等真菌有明显的抑制作用.A chitinase with antifungal activity was isolated from adzuki bean （ Vigna angularis） seeds by a procedure containing （NH4）2SO4 precipitation, ionexchange chromatography on CM -Sepha- dex C -50, gel filtration on Sephadex G- 75, and high-performance liquid chromatography （HPLC） on Poros HS - 20. The protein exhibited a molecular mass of 27.7 kDa in reduced condition while 22.7 kDa in oxidized condition on SDS - PAGE, which indicates that there are disulfide bonds inside the molecule. The specific activity of the chitinase was estimated to be 14.57 U/mg. The enzyme expressed its optimum activity at pH 5.4 and a temperature of 50 ℃. It exerted obvious antifungal action towards Fusarium oxysporum, Fusarium oxysporum f. sp. nelonis, and Pythium aphanidermatum.

关 键 词：红豆 几丁质酶 纯化 表征 

分 类 号：Q814.1[生物学—生物工程]