摘要
斑玉蕈子实体经生理盐水抽提、30%～60%饱和度的硫酸铵沉淀、DEAE-Cellulose和Sephadex G-100柱层析纯化得到斑玉蕈凝集素（Hypsizygus marmoreus Lectin,简称HML）.HML经PAGE显示单一条带,SDS-PAGE测得亚基相对分子质量为34.2 kD,SephadexG-100凝胶过滤测得相对分子质量为35 kD,中性糖含量为7.2%,含有17种氨基酸,IEF-PAGE测得其等电点为8.15.该凝集素能凝集多种动物红细胞和人的A、B、AB和O血型红细胞,对兔红细胞的凝集作用可被甘露糖、半乳糖、N-乙酰半乳糖胺和岩藻糖所抑制.HML对热较敏感,经50℃处理10 min,凝集活性下降明显,但对酸碱具有一定的稳定性,在pH 5.0～8.0范围内较稳定.β-消去反应测得其糖和蛋白质的连接键为O-型糖肽键.
The lectin HML was isolated and purified from the fruiting bodies of Hypsizygus marmoreus by physiological saline extraction, precipitation of 30--60% （NH4）2SO4 and DEAE-Cellulose, and Sephadex G-100 chromatography. It turned out to be a single band in PAGE identification. The relative molecular mass of HML was 35 kD tested by gel filtration, and the subunit of HML was 34.2 kD determined by SDS-PAGE. HML contained 7.2% neutro-saccharide, and contained 17 kinds of amino acid. Its pI was about 8.15. This lectin agglutinated erythrocytes of the animal and human beings of A, B, AB and O blood groups, the hemagglutination of the lectin on rabbit erythrocytes was inhibited by manose, galactose, GalNAc and fucose among the sugars tested. HML was sensitive to heat, and the agglutinating activity declined obviously after being heated 10 minutes under 50 ℃. It possessed high activity within pH 5.0-8.0. β-Elimination revealed the bond between the polysaccharide and the protein of HML was O-type glucopeptide one.
出处
《菌物学报》
CAS
CSCD
北大核心
2006年第2期284-291,共8页
Mycosystema
基金
福建省教育厅资助项目(No.JB00029)
福建省高校测试基金资助项目(No.CJ-120)
关键词
食用真菌
生理活性物质
糖蛋白
纯化
edible fungi
physiological active substance
glycoprotein
purification
