International Journal of Molecular Sciences | |
Purification and Characterization of Iso-Ribonucleases from a Novel Thermophilic Fungus | |
Kyle S. Landry1  | |
[1] Department of Food Science, Massachusetts Agricultural Experiment Station, University of Massachusetts, Amherst, MA 01003, USA; E-Mail | |
关键词: affinity purification; RNase purification; chromatography; thermophilic fungi; RNase characterization; | |
DOI : 10.3390/ijms15010944 | |
来源: mdpi | |
【 摘 要 】
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.
【 授权许可】
CC BY
© 2014 by the authors; licensee MDPI, Basel, Switzerland
【 预 览 】
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RO202003190029906ZK.pdf | 921KB | download |