芝麻菜种子抗菌蛋白的纯化及其抗菌活性

张耀; 王财成; 袁素素; 张玮玮; 叶秀娟

doi:10.13386/j.issn1002-0306.2018.20.016

芝麻菜种子抗菌蛋白的纯化及其抗菌活性

Purification of Antibacterial Protein from Arugula (Eruca sativa Mill.) Seeds and Its Antifungal Activity

摘要

摘要: 本文旨在从芝麻菜种子中分离纯化得到一种具有抗菌活性的蛋白,并对其抗菌活性进行研究。通过SP-Sepharose离子交换色谱、Mono S^TM 5/50GL和Superdex^TM 75 10/300GL快速蛋白液相色谱进行分离纯化,经Tricine-SDS-PAGE电泳分析;采用纸片扩散法、荧光染色法,对其抗菌活性和稳定性进行研究。结果表明,从芝麻菜种子中纯化一种抗菌蛋白ZSU2,经Tricine-SDS-PAGE电泳鉴定达到电泳纯,相对分子质量约为12 kDa;ZSU2蛋白对小白菜炭疽病菌等12种真菌具有较强的抑制作用,抗菌活性在pH1~13、温度0~100℃稳定,且具有较强的金属阳离子耐受能力;此外,ZSU2蛋白能引起菌丝尖端几丁质积累和细胞膜选择透过性的改变。

Abstract: This essay was aimed to isolate, purify and characterize the protein with antifungal activity from arugula (Eruca sativa Mill.) seeds.The antifungal protein was isolated and purified from the seed of arugula by ion exchange chromatography on SP-Sepharose and followed by fast protein liquid chromatography on Mono S^TM 5/50 GL and Superdex^TM 75 GL.Tricine-SDS-PAGE was used to analysis its purity and molecular weight.The antifungal activity and stability were studied by means of paper diffusion and fluorescence staining.The results showed that, the antifungal protein (ZSU2) isolated and purified from the seed of arugula exhibited a molecular mass about 12 kDa in Tricine-SDS-PAGE electrophoresis.It had a strong inhibitory effect on 12 kinds of fungi, such as Colletotrichum higginsianum. The antifungal activity was stable on pH 1 to 13 and temperature 0 to 100℃, and had a strong metal cation tolerance.In addition, ZSU2 protein could induce chitin accumulation and cell membrane selection through sexual change.