Purification of an Inducible DNase from a Thermophilic Fungus
1
Department of Food Science, Massachusetts Agricultural Experiment Station, University of Massachusetts, Amherst, MA 01003, USA
2
The Stockbridge School of Agriculture, University of Massachusetts, Amherst, MA 01003, USA
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2014, 15(1), 1300-1314; https://doi.org/10.3390/ijms15011300
Received: 6 December 2013 / Revised: 6 January 2014 / Accepted: 9 January 2014 / Published: 20 January 2014
(This article belongs to the Special Issue Thermophilic DNases, RNases and Proteases)
The ability to induce an extracellular DNase from a novel thermophilic fungus was studied and the DNAse purified using both traditional and innovative purification techniques. The isolate produced sterile hyphae under all attempted growing conditions, with an average diameter of 2 µm and was found to have an optimal temperature of 45 °C and a maximum of 65 °C. Sequencing of the internal transcribed region resulted in a 91% match with Chaetomium sp., suggesting a new species, but further clarification on this point is needed. The optimal temperature for DNase production was found to be 55 °C and was induced by the presence of DNA and/or deoxyribose. Static growth of the organism resulted in significantly higher DNase production than agitated growth. The DNase was purified 145-fold using a novel affinity membrane purification system with 25% of the initial enzyme activity remaining. Electrophoresis of the purified enzyme resulted in a single protein band, indicating DNase homogeneity.
View Full-Text
Keywords:
affinity purification; DNase purification; chromatography; thermophilic fungi; affinity membrane purification
This is an open access article distributed under the Creative Commons Attribution License
MDPI and ACS Style
Landry, K.S.; Vu, A.; Levin, R.E. Purification of an Inducible DNase from a Thermophilic Fungus. Int. J. Mol. Sci. 2014, 15, 1300-1314.
AMA Style
Landry KS, Vu A, Levin RE. Purification of an Inducible DNase from a Thermophilic Fungus. International Journal of Molecular Sciences. 2014; 15(1):1300-1314.
Chicago/Turabian StyleLandry, Kyle S.; Vu, Andrea; Levin, Robert E. 2014. "Purification of an Inducible DNase from a Thermophilic Fungus" Int. J. Mol. Sci. 15, no. 1: 1300-1314.
Find Other Styles
Search more from Scilit