黑芸豆中新型凝集素的分离纯化及其功能活性  

Isolation and Purification of a Novel Lectin from Black Kidney Bean and Its Functional Properties

作　　者：蔡茜茜[1] 洪永祥[1] 汪少芸[1] 

机构地区：[1]福州大学生物科学与工程学院,福州350108

出　　处：《中国食品学报》2015年第6期96-101,共6页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(31471623);福州市科技项目(2014-G-62);福清市科技项目(融科2014-23)

摘　　要：目的：分离纯化一种黑芸豆凝集素,对其部分生物活性进行表征和研究。方法：采用缓冲液抽提、AFBlue HC-650M亲和色谱、CM Sephadex c-25阳离子交换色谱及Sephadex G-50凝胶过滤色谱进行分离纯化,通过SDS-PAGE分析鉴定,并对其凝血活性、抗真菌活性进行表征。结果：从黑芸豆中分离纯化出一种凝集素,命名为BKBL,经SDS-PAGE鉴定达到电泳纯。BKBL为单体蛋白,相对分子质量为36 ku,经PAS染色鉴定为糖蛋白,含糖量11.9%。其对人血和小鼠血均具有一定的凝集活性,对小鼠血的凝集活性高于人血。BKBL凝血活性在70℃以下,p H 2~10范围内高度稳定,在p H 11处理30 min后活性保留50%,在100℃处理30 min后活性完全丧失。抑菌试验表明,BKBL对黑曲霉、香蕉尖孢镰刀菌、锥栗炭疽病病菌、莲细极链格孢菌、瓜果腐霉病菌、葡萄灰霉菌具有抑制作用。结论：从黑芸豆分离纯化出的一种凝集素,对小鼠血有较高的凝集活性,性质稳定;对多种真菌有抑制作用。Objective： The main purpose of this research is isolation, purification and partial characterization of a lectin from black kidney beans. Methods： The isolation and purification procedure contained buffer extraction, affinity chromatography on AF-Blue HC-650M, ion-exchange chromatography on CM Sephadex c-25, gel filtration chromatography on Sephadex G-50. SDS-PAGE was used to identify its purity and analyze the molecular weight, and the partial bi- ological function activity of the lectin including hemagglutinating activity and antifungal activity had also been studied.Results： A lectin was isolated and purified from Black kidney beans, named BKBL. The lectin was a purified monomeric protein with a molecular weight of 36 ku by SDS-PAGE, and was also identified as a glycoprotein with 11.9% sugar content through PAS dyeing. BKBL exhibited hemagglutinating activity in both human erythrocytes and mice erythrocytes, and the effect was more obvious in the latter. The hemagglutinating activity was not effected below 70 ℃ and between pH 2-10, its activity remained 50% after being treated with pH 11 for 30 min, but it would lose its activity at 100 ℃ treatment for 30 rain. BKBL showed antifungal activity towards the growth of AspergiUus niger, Fusarium oxysporum f.sp. cubense, Colletotrichum gloeosporioides Penz, Alternaria tenuissima, Pythium aphanidermatum, Botrytis einerea. Conclusion： A lectin can be isolated and purified from Black kidney beans, with strong and stable hemagglutinating activity to- wards mice erythrocytes, and antifungal activity towards most fungi.

关 键 词：黑芸豆 凝集素 凝血 抗真菌 

分 类 号：TS214[轻工技术与工程—粮食、油脂及植物蛋白工程]