【摘要】 鸡菌丝体浸取液依次经硫酸铵分级沉淀,DEAE-Sepharose CL-6B离子交换层析和Sephadex G-100分子筛层析3个主要步骤纯化得到一种凝集素(Termitomyces albu-minosus lectin,简称TAL)。纯化的TAL在聚丙烯酰胺凝胶电泳上显示一条蛋白质着色带。TAL的分子量为89.4kD,亚基分子量为38kD和51kD,提示TAL分子由两个不同亚基组成。TAL具有供血动物种属专一性,使Wistar大鼠红细胞凝集所需TAL最低的浓度为0.49μg/ml。糖抑制试验表明,鸡卵粘蛋白明显抑制TAL的凝血活性。TAL对热不稳定,60℃保温15min活力完全丧失。钙、镁或锰离子对TAL无激活作用。TAL不含不性糖,Glu和Asp含量较高,His和Met含量较低。更多还原

【Abstract】 Termitomyces albuminosus lectin (TAL) has been purified by extracting of mycelia prepared by submerged culture with PBS,ammonium sulfate fractionation,DEAE-Sepharose ion-exchange chro-matography and Sephadex G-100 gel filtration. The purified lectin showed a single band on PAGE in pH8. 9 buffer. This lectin was composed of two different subunits of 38 and 51kD determined by SDS-PAGE and the molecular mass of the intact lectin was estimated to be 89. 4kD by gel filtration. TAL was specific in agglutination for erythrocytes of Wistar rats and SD rats. The minium concentration required for hemagglutination was 0. 49μg/mL and 3. 9μg/mL respectively. The hemagglutination activity of TAL could be inhibited specifically by Chicken Ovomucin. TAL was heat unstable and lost all activity at 60℃ for 15 min. Metal cations Ca2+ ,Mg2+ and Mn2+ had no effects on TAL. TAL contained no neutral sugar determined by the phenol-H2SO4 method with reference to glucose. The amino acid composition of TAL was also etermined.更多还原