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Int. J. Mol. Sci. 2014, 15(1), 944-957; doi:10.3390/ijms15010944
Article

Purification and Characterization of Iso-Ribonucleases from a Novel Thermophilic Fungus

 and *
Received: 10 December 2013; in revised form: 31 December 2013 / Accepted: 2 January 2014 / Published: 10 January 2014
(This article belongs to the Special Issue Thermophilic DNases, RNases and Proteases)
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Abstract: A thermophilic fungus previously isolated from composted horse manure was found to produce extracellular iso-RNases that were purified 127.6-fold using a combination of size exclusion chromatography and a novel affinity membrane purification system. The extent of purification was determined electrophoretically using 4%–15% gradient polyacrylamide gels. RNase activity was dependent on the presence of a metal co-factor with significantly more activity with Zn2+ or Mn2+ than Mg2+. The RNases exhibited maximum activity at both pH 3.0 and pH 7.0 with no activity at pH 2.0 or 10.0. The optimal temperature for the iso-RNase was 70 °C. The molecular weight of the iso-RNase was determined to be 69 kDa using a Sephadex G-75 column.
Keywords: affinity purification; RNase purification; chromatography; thermophilic fungi; RNase characterization affinity purification; RNase purification; chromatography; thermophilic fungi; RNase characterization
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Landry, K.S.; Levin, R.E. Purification and Characterization of Iso-Ribonucleases from a Novel Thermophilic Fungus. Int. J. Mol. Sci. 2014, 15, 944-957.

AMA Style

Landry KS, Levin RE. Purification and Characterization of Iso-Ribonucleases from a Novel Thermophilic Fungus. International Journal of Molecular Sciences. 2014; 15(1):944-957.

Chicago/Turabian Style

Landry, Kyle S.; Levin, Robert E. 2014. "Purification and Characterization of Iso-Ribonucleases from a Novel Thermophilic Fungus." Int. J. Mol. Sci. 15, no. 1: 944-957.


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