Purification of an Inducible DNase from a Thermophilic Fungus

• Main Authors: Kyle S. Landry, Andrea Vu, Robert E. Levin
• Format: Article
• Language: English
• Published: MDPI AG 2014-01-01
• Series: International Journal of Molecular Sciences
• Subjects: affinity purification; DNase purification; chromatography; thermophilic fungi; affinity membrane purification
• Online Access: http://www.mdpi.com/1422-0067/15/1/1300

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.