文章摘要
李丹彤.刺参凝集素的分离纯化及其性质[J].水产学报,2005,29(5):
刺参凝集素的分离纯化及其性质
Isolation, purification and properties of lectin from Apotichopus japonicus
投稿时间:2014-03-10  修订日期:2014-03-10
DOI:
中文关键词: 刺参 凝集素 纯化 性质
英文关键词: Apotichopus japonicus lecfin purification properties
基金项目:国家自然科学基金
作者单位地址
李丹彤 大连水产学院农业部海洋水产增养殖学与生物技术重点开放实验室 大连水产学院农业部海洋水产增养殖学与生物技术重点开放实验室 辽宁大连116023 大连水产学院农业部海洋水产增养殖学与生物技术重点开放实验室; 大连水产学院农业部海洋水产增养殖学与生物技术重点开放实验室 辽宁大连116023
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中文摘要:
      为探寻刺参药用价值,探索其抑制肿瘤细胞增殖,促进淋巴细胞转化等生物功能.将刺参(Apotichopus japonicus)经磷酸盐缓冲溶液抽提,20%~75%硫酸铵分级沉淀,Sephadex G-200分子筛层析,得到刺参凝集素(AJL).用Sephadex G-200分子筛层析测得其分子量为85 652 Da.AJL的含糖量为20.9%.该凝集素可以凝集人的A、B、AB、O型红细胞,且对A型红细胞凝集作用最强, 在对兔、鼠、鲤鱼、鸡和人的A、B、AB、O红细胞的凝集作用中,对鸡的凝集作用最强.该凝集素凝集兔红细胞的作用不被D-果糖、D-甘露糖、γ-球蛋白、葡萄糖、蔗糖、甘露聚糖所抑制,而被牛甲状腺球蛋白所抑制,其最小抑制浓度为1.55 mg?mL-1.该凝集素凝集兔红细胞的作用不被二价金属离子Ca2 、Mg2 及EDTA所抑制.该凝集素在pH 4.0~10.14系列缓冲液中均有活性,其中在pH 4.4~7.5范围内血凝活性最高.该凝集活性在90 ℃加热30 min后仍然对红细胞有凝集活性,显示出较高的热稳定性.
英文摘要:
      Lectin was purified from Apotichopus japonicus Selenka by extraction with PBS, followed by 20% - 75% ammonium sulfate fractionation and molecular sieve chromatography on the Sephadex G-200 . The Lectin had a molecular weight of 85 652 Da on Sephadex G-200 and contained saccharied of 20.9%. The lectin was nonspecific in agglutination for any type of human erythrocytes, and the hemagglutinating activity for human( A) was the highest, the agglutination for chicken was the highest, among erythrocytes of humans( A, B, AB, O), rabbits, mouse and common carp. The hem,agglutinating activity for rabbit could not be inhibited by D-galactose, glucose, sucrose, mannan, gamma-globulin(human), but inhibited by bovine-thyroglobulin. The minimum inhibitory concentration was 1.55 mg?mL^-1. The hemagglutinating activity was not inhibited by treatment with EDTA and Ca^2+ ,Mg^2+ . This lectin had the maximum hemagglutinating activity at pH 4.4 - 7.5, with activity at pH 4.0 - 10.14. The lectin still showed aggllutinating activity after being treated with 90℃ for 30 minutes, indicating that it had a high heat resistance.
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