中华绒螯蟹血清凝集素的分离纯化与性质研究  

Purification and Characteristics of Agglutinin in Serum of Chinese Mitten Crab Eriocheir sinensis

作　　者：徐海圣[1] 徐步进[2] 

机构地区：[1]浙江大学动物科学学院,浙江杭州310029 [2]浙江大学原子核农业科学研究所,浙江杭州310029

出　　处：《水产科学》2008年第9期455-461,共7页Fisheries Science

基　　金：浙江省科技计划项目(2004C32050)

摘　　要：运用硫酸铵盐析和GlcNAc-Sepharose6B亲合层析等方法从中华绒螯蟹的血清中分离出一种天然的凝集素，经SDS-PAGE测得其分子量约为82kD，由单一亚基组成，由等电聚焦凝胶电泳测得其等电点（pI）为5．4。糖凝集抑制试验中检测到葡萄糖、半乳糖、甘露糖、乳糖、果糖和蔗糖对中华绒螯蟹血清凝集素均没有抑制作用，但N-乙酰氨基糖GlcNAc、GalNAc和ManNAc等能抑制其凝集活性。热变性试验结果表明，温度为10～50℃，凝集素仍保持强的凝集活性，但温度升至60℃后，凝集活性迅速下降，至80℃失去凝集活性，在pH4．0～8．0的各缓冲液中保持较强的凝集活性，而在此pH范围外，凝集活性均有不同程度的下降。中华绒螯蟹血清凝集素可使外源红血细胞发生凝集，还具有凝集细菌或抗菌作用。A natural agglutinin from the serum of Chinese mitten crab, Eriocheir sinensis, was purified by affinity chromatography on GlcNAc-Sepharose 6B. The purified agglutinin was composed of one subunit, the molecular weight was 82 kDa by SDS - PAGE, and its isoelectric point was 5.4. The agglutinating inhibition test showed that the purified agglutinin was not be inhibited obviously by glucose, galactose, mannose, lactose, fructose, and sucrose, but N-acetylglucosamine （ GlcNAc）, N-acetylgalactosamine （GalNAc） and N-acetylmannosamine （ManNAc） had obvious inhibited effect on the agglutination. The agglutinating activity was strong and stable at water temperature of 10 -50℃ ,and declined dramatically when temperature inclined to 60℃ and losed its activity when temperature inclined to 80℃. The agglutinin also showed an optimum hemaglutinating activity over a range of pH 4.0 ～ 8.0. The agglutinin from the serum of the Chinese mitten crab had antibacterial activity.

关 键 词：中华绒螯蟹 凝集素 纯化 性质 

分 类 号：Q959.223[生物学—动物学]