ORIGINAL ARTICLES   

Minerva Biotecnologica 2013 September;25(3):165-70

Copyright © 2013 EDIZIONI MINERVA MEDICA

language: English

Production, purification and biochemical analysis of RNase producing mutant strain of Streptomyces venezuelae

Sunhare R. ^{1, 2}, Punniyamoorthy G. ³, Prabu R. ^{2, 4} ✉

¹ Institute of Bioscience, Universiti Putra Malaysia, UPM Serdang, Selangor, Malaysia; ² School of Biotechnology, University Institute of Technology, Rajiv Gandhi Proudyogiki Vishiwavidyalaya, Bhopal, India; ³ Department of Biochemistry, M.R Government College, Mannargudi, India; ⁴ Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM Serdang, Selangor, Malaysia

Aim: The aim of this study was to improve the intracellular and extracellular RNase producing Streptomyces venezuelae strain.
Methods: The random mutation was subjected into Streptomyces venezuelae with N-Methyl-N’ nitro-N-nitrosoguanidine (NTG) to increase the RNase enzyme production. The presence of the enzyme was confirmed by RNase activity method and Native-PAGE electrophoresis. This RNase R family was purified by multistep purifications with cation-exchange chromatography and size exclusive chromatography respectively.
Results: There was 3-4fold increment in RNase R enzyme has observed with selected N-Methyl-N’ nitoro-N-nitrosognanidine (NTG) mutated clones. The purified RNase shows stability against various metal ions with enzyme activity at pH 7 and temperature 37°C.
Conclusion: These approaches are able to resolve the contradiction between the low production and high demand of RNase enzyme.