Zhang Rui, Tian Guoting, Zhao Yongchang, Zhao Liyan, Wang Hexiang, Gong Zhiyuan, Ng Tzi Bun
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, China.
J Basic Microbiol. 2015 Feb;55(2):269-75. doi: 10.1002/jobm.201300876. Epub 2014 Feb 12.
A purification protocol that encompassed anion exchange chromatography (EC) on DEAE-cellulose, cation EC on CM-cellulose, anion EC on Q-Sepharose, and fast protein liquid chromatography-gel filtration of Superdex 75 was used to isolate a ribonuclease from dried fruiting bodies of Ramaria formosa. The ribonuclease was unadsorbed on CM-cellulose but adsorbed on both DEAE-cellulose and Q-Sepharose. It displayed a molecular mass of 29-kDa in both gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The ranking of its ribonucleolytic activity toward polyhomoribonucleotides was poly(U) > poly(C) > poly(G) > poly(A). It exhibited a pH optimum of pH 5 and a temperature optimum at 60 °C. The ribonuclease inhibited HIV-1 reverse transcriptase with an IC50 of 3 µM.
采用一种纯化方案从美丽枝瑚菌的干燥子实体中分离核糖核酸酶，该方案包括在DEAE-纤维素上进行阴离子交换色谱（EC）、在CM-纤维素上进行阳离子EC、在Q-琼脂糖上进行阴离子EC以及Superdex 75的快速蛋白质液相色谱-凝胶过滤。该核糖核酸酶不吸附于CM-纤维素，但吸附于DEAE-纤维素和Q-琼脂糖。在凝胶过滤和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中，其分子量均为29 kDa。它对多聚同型核糖核苷酸的核糖核酸酶活性排序为：聚（U）＞聚（C）＞聚（G）＞聚（A）。其最适pH为5，最适温度为60℃。该核糖核酸酶抑制HIV-1逆转录酶，IC50为3 μM。
