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磷沙蚕凝集素的分离纯化及生物活性研究
孔 静1, 熊川男1, 李 伟1,2, 孔 亮1, 金 桥2, 白雪芳1, 杜昱光1
1.中国科学院大连化学物理研究所生物技术部;2.大连水产学院食品工程系, 辽宁省水产品加工与贮藏重点开放实验室
摘要:
采用DEAE-Toyopearl阴离子交换树脂、Bio- Gel P-100分子筛层析、Lactosyl-Sepharose4B亲和层析和Sephadex 75分子筛层析技术从日本海的海洋无脊椎动物磷沙蚕(Chaetopterus variopedatus)中分离出2种凝集素,通过体外细胞活性和酶活性实验观察它们的生物活性。结果表明,从海洋无脊椎动物磷沙蚕中分离出一种β-半 乳糖特异性凝集素(CVL)和一种具有血凝活性,但不被经测试的单糖和糖蛋白所抑制的相对分子质量为30 000的蛋白质(CVL-1)。β-半乳糖特异性凝集素(CVL)具有体外抑制NB4白血病细胞株的活性和抑制小鼠脾细胞的活性,IC50分别为7.87 mg/L和22 mg/L。同时,另一种蛋白质(CVL-1)具有β-葡萄糖苷酶活性。
关键词:  磷沙蚕(Chaetopterus variopedatus)  凝集素  β-葡萄糖苷酶  NB4白血病细胞株  脾细胞
DOI:
分类号:
基金项目:辽宁省自然科学基金项目(2050885);辽宁省教育厅资助项目(05L086);大连水产学院科研项目(015690)
The lectins from sea worm Chaetopterus variopedatus: isolation and biological activity
Abstract:
A β-galactose-specific lectin(CVL) was purified from the sea worm Chaetopterus variopedatus. CVL displays cytotoxic effect on NB4 human leukemia cells and T, B lymphocytes of mouse. The highest limit of CVL concentration causing a decrease in cell survival of 50% was evaluated to be 7.87 mg/L for NB4 cells and 22 mg/L for T, B lymphocytes of mouse. A new lectin(CVL-1) was purified from the same sea worm. Molecular weight of CVL-1 obtained was determined by SDS-PAGE to be 30 000. The hemmagglutination activity of CVL-1 was not inhibited by any of the monosaccharides and glycoproteins tested. CVL-1 is presented with β-glucosidase activity.
Key words:  Chaetopterus variopedatus  lectin  β-glucosidase  NB4 human leukemia cell  lymphocyte
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