Studies on the toxin of Aspergillus fumigatus. VII. Purification and some properities of hemolytic toxin (asp-hemolysin) from culture filtrates and mycelia.
Studies on the toxin of Aspergillus fumigatus. VII. Purification and some properities of hemolytic toxin (asp-hemolysin) from culture filtrates and mycelia.
复制标题
烟曲霉毒素的研究.
DOI：
10.1016/j.ejphar.2015.06.062
复制
发表时间：
1977
影响因子：
2.6
通讯作者：
O. Sakaguchi
中科院分区：
文献类型：
--
作者：
K. Yokota;H. Shimada;A. Kamaguchi;O. Sakaguchi
通过硫酸铵沉淀、DEAE-Sephadex 层析、伴刀豆球蛋白 A-Sepharose 亲和层析、Sephadex G-50、G-100 和 G-150 凝胶过滤，从烟曲霉菌丝体和培养滤液中获得溶血毒素。纯化的均溶毒素在免疫学和圆盘电泳分析中是均质的，并且通过免疫扩散技术从培养滤液中得到的毒素与从菌丝体中得到的毒素相同。该溶血毒素首次从真菌中分离得到，命名为Asp-溶血素。通过凝胶过滤技术估计Asp-溶血素的分子量约为30,000，发现其等电点约为pH 4.0。这种天冬氨酸溶血素含有大量的蛋白质和极少量的碳水化合物。 Asp-溶血素的紫外吸收光谱在280 nm处有最大吸收，在251 nm处有最小吸收。 280 nm 处的消光系数和 251 nm 处的最小吸收。 280 nm（E 1% 1CM）处的消光系数为12.4，280 nm处与260 nm处的吸光度之比为2.3。室温下该毒素对鸡红细胞溶血活性的最适pH为5.0，在pH 3.5～10.5范围内有活性。最适温度为21℃，50℃孵育5分钟或45℃孵育23分钟约50％的活性损失。 Hg2+、Cu2+、Fe2+、Ag1+、碘和p-CMB显着抑制溶血活性，而Zn2+和Co2+则略有增强溶血活性。
A hemolytic toxin has been obtained from mycelia and culture filtrates of Aspergillus fumigatus by the procedures that included precipitation with ammonium sulfate, chromatography of DEAE-Sephadex, affinity chromatography on Concanavalin A-Sepharose and gell filtration on Sephadex G-50, G-100 AND G-150. The purified homolytic toxin was homogeneous on immunological and disk electrophoretic analysis, and the toxin from culture filtrates was identical with that from mycelia by the immunodiffusion technique. The hemolytic toxin was obtained for the first time from fungi and designated as Asp-hemolysin. The molecular weight of Asp-hemolysin was estimated to be appoximately 30,000 by the gel-filtration technique and its isoelectric point was found to be around pH 4.0. This Asp-hemolysin contained large amounts of protein and very small amounts of carbohydrate. The UV absorption spectrum of Asp-hemolysin showed a maximum absorption at 280 nm and minimum absorption at 251 nm. The extinction coefficient at 280 nm and minimum absorption at 251 nm. The extinction coefficient at 280 nm, E 1% 1CM, was 12.4 and the ratio of absorbance at 280 nm to that at 260 nm was 2.3. The optimum pH for the hemolytic activity of the toxin toward chicken erythrocytes was 5.0 at room temperature and it was active in the pH range of 3.5 to 10.5. The optimum temperature was 21 C and about 50% of the activity was lost by incubation at 50 C for 5 min or 45 C for 23 min. The hemolytic activity was remarkably inhibited by Hg2+, Cu2+, Fe2+, Ag1+, iodine and p-CMB, but enhanced slightly by Zn2+ and Co2+.