祁连山荷叶离褶伞菌丝体LML的分离纯化及其凝血活性检测

Purification and hemagglutinating activity test of the lectin isolated from mycelium of Lyophyllum decastes from Qilian mountain

以荷叶离褶伞茵丝体为材料，利DEAE—Sepharose F．F．色谱和sephadexG-100色谱技术，分离获得新的LMLLML。根据凝胶过滤色谱和SDS—PAGE电泳结果，确定LML是中性糖含量为10．36％、分子量约为48．2ku单亚基糖蛋白。凝血活性检测结果表明，LML对兔血红细胞具有凝聚活性，这种活性在LML浓度为500μg/mL、温度为20～70℃、pH3～10的条件下稳定；其凝血活性可被葡萄糖（最小浓度为25mmol／L）和麦芽糖（最小浓度为12．5mmol／L）所抑制；当LML浓度≥250μg／mL时，其凝血活性不受Ca^2＋、Mg^2＋和Zn^2＋的影响。

One novel lectin LML was purified from mycelium of Lyophyllum decastes using ion exchange chromatography and gel filtration. Based on the result of gel filtration and SDS-PAGE,LML was single subunit glycoprotein with a molecular mass of 48.2ku and having 10.36% neutral sugar. The hemagglutinating activity test showed that LML showed hemagglutinating activity on rabbit red blood cells. Its activity was stable in the pH range of 3-10,temperature range of 20-70℃, The hemagglutinating activity of LML were inhibited by glucose （25mmol/L） and maltose （12.5mmol/L） ,and were unaffected by Ca2＋, Mg2＋ and Zn2＋ at LML concentrition≥ 250μg/m L.