Premium
A novel ribonuclease with HIV‐1 reverse transcriptase inhibitory activity purified from the fungus Ramaria formosa
Author(s) -
Zhang Rui,
Tian Guoting,
Zhao Yongchang,
Zhao Liyan,
Wang Hexiang,
Gong Zhiyuan,
Ng Tzi Bun
Publication year - 2015
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.201300876
Subject(s) - ribonuclease , chromatography , size exclusion chromatography , chemistry , polyacrylamide gel electrophoresis , sepharose , molecular mass , sodium dodecyl sulfate , cellulose , gel electrophoresis , ion chromatography , biochemistry , enzyme , rna , gene
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 IC 50 of 3 µM.