姬松茸凝集素的分离纯化及性质  

Purification and characterization of lectin from Agaricus blazei Murrill

作　　者：王文君 王璐瑶[1] 周华[1] 韦萍[1] WANG Wenjun;WANG Luyao;ZHOU Hua;WEI Ping(College of Biotechnology and Pharmaceutical Engineering,Nanjing Tech University,Nanjing 211800,China)

机构地区：[1]南京工业大学生物与制药工程学院,江苏南京211800

出　　处：《生物加工过程》2020年第3期318-323,共6页Chinese Journal of Bioprocess Engineering

基　　金：国家重点基础研究计划(973计划)(2013CB733904)。

摘　　要：凝集素是一类能够识别特异性糖并与之非共价结合的蛋白或糖蛋白,因其具有特定的识别受体,故在免疫系统和发育过程中发挥了重要的作用。使用DEAE-Sepharose FF弱阴离子交换层析、SP-Sepharose强阳离子交换柱层析、Sephadex G-75分子筛凝胶层析对姬松茸凝集素进行分离纯化,研究理化因素对其凝血活性的影响及其抗菌活性。结果表明:纯化的样品经SDS-PAGE电泳分析得到单一条带,姬松茸凝集素是含有2个相同亚基的同源二聚体,其分子量都为1.20×10^5。纯化后的姬松茸凝集素相对粗提液的回收率为28.99%,纯化倍数为19.06倍。姬松茸凝集素对热有较好的耐受性,但温度为90℃时失去凝血活性;在强酸或强碱下丧失活性,最适pH为7~9;凝集活性不受金属离子的影响;葡萄糖是其专一性结合糖。姬松茸凝集素对褐腐菌的生长起到了明显的抑制作用,而对大肠杆菌、枯草芽孢杆菌、绿色木霉、西瓜枯萎病菌和黑曲霉的生长没有明显的影响。Lectins are a kind of proteins or glycoproteins that interact with carbohydrates.Lectins can function as pattern recognition receptors and play an important role in the innate immune system and development.We isolated a lectin from the edible mushroom Agaricus blazei Murrill(ABM)by DEAE-Sepharose FF anion exchange chromatography,SP-Sepharose strong cation-exchange chromatography and Sephadex G-75 gel filtration chromatography.The physicochemical properties and antibacterial activity of the ABM lectin were also studied.The results show that the obtained lectin possessed one subunit using SDS-PAGE.It contained two homologous dimers of the same subunit with the molecular weight of 1.20×10^5.The recovery yield of the purified lectin from ABM was 28.99% and the purification ratio was 19.06 times.The ABM lectin had good resistance to heat when the temperature is below 90℃,otherwise it lost cruor activity.When it was in the extreme acid or extreme alkali environment,the activity of lectin lost and the range of optimum pH was 7-9.Its agglutination activity did not depend on metal ions.Glucose was its specific binding sugar.ABM lectin had obvious inhibitory effect on the growth of brown rot fungi and had no significant effect on the growth of Escherichia coli,Bacillus subtilis,Trichoderma viride,Fusarium oxysporum f.sp.niveum and Aspergillus niger.

关 键 词：姬松茸 凝集素 凝血活性 抑菌活性 

分 类 号：Q513[生物学—生物化学]