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泥鳅血清凝集素的研究
姚东瑞1,2, 盘赛昆1, 周鸣谦1,2, 葛荣荣1
1.淮海工学院海洋学院;2.江苏海洋资源研究院
摘要:
采用DEAE-Sepharose FF离子交换层析和Superdex 200 10/300 GL凝胶过滤层析技术,从泥鳅血清中分离出两种天然凝集素MSL-I和MSL-。SDS-PAGE检测表明,MSL-I及MSL-Ⅱ均显示两条蛋白质着色带。采用凝胶过滤法测得MSL-I相对分子质量为25 000,MSL-Ⅱ相对分子质量为82 000。根据凝胶过滤测定及SDS-PAGE检测结果可知,MSL-Ⅱ由相对分子质量分别为29 000和13 600两种亚基组成,MSL-I的亚基组成有待进一步研究。MSL-Ⅱ能结合多种糖,其中对乳糖的亲和力最强。MSL-Ⅱ具有较强的热稳定性,在pH 4~11范围内具有凝集活性,最适pH为6~8。
关键词:  泥鳅  血清  凝集素  分离纯化
DOI:
分类号:
基金项目:江苏省科技支撑计划(农业)项目(BE2010399)、江苏省农业科技自主创新基金项目[CX09-627;CX(10)328]和连云港市科技基础设施建设计划项目(CK0935)共同资助
Preliminary study on lectins from serum of loach Misgurnus anguillicaudatus
Abstract:
Two lectins with different molecular mass,designated as MSL-I and MSL-II, were isolated from serum of loach Misgurnus anguillicaudatus. The purification procedure consisted of separation on DEAE-Sepharose FF anion exchange column followed by gel filtration chromatography on Superdex 200 10/300 GL. Determination showed that MSL-I and MSL-II had a nature molecular weight of 25,000 and 82,000 respectively.The denatured SDS-PAGE showed that both MSL-I and MSL-II had two bands on the gel. We proposed that MSL-II was constituted with two subunits with molecular mass of 29,000 and 13,600 respectively. The subunit composition of MSL-I was to be further studied. Sugar-binding specificity test showed that glucose, D-galactose, D-fructose, D-xylose, L-arabinose and lactose had inhibitory effect on MSL-II, and among them lactose was most effective.Thermal stability test showed that MSL-II had a strong thermal stability even treated at 60℃ for 30min. The optimum pH of MSL-II was 6~8.
Key words:  Loach Misgurnus anguillicaudatus  Serum  Lectin  Purification