Production and Characterization of partial purified Protease from the fungal species Trichoderma harzianum | ||
| journal of kerbala university | ||
| Article 1, Volume 8, Issue 2, June 2012, Pages 121-132 PDF (0 K) | ||
| Abstract | ||
| Laboratory experiment was achieved for production and partial purification of protease from the fungal species Trichoderma harzianum. After screening of enzyme productivity by calculating the radial of hemolysis zone in Skimed milk was 7 cm. The optimal conditions for protease production were studied using liquid state fermentation composed of wheat bran as protein source. The results revealed the optimal conditions of enzyme production after five days of incubation at 25 C in liquid mineral medium with 2% concentration of wheat bran and pH 4.0. The protease was purified by several steps, including precipitation with 80 % saturation of ammonium sulphate as the first step of purification, the obtained purification fold was 3.47 and recovery 43%. Second step of partial purification which was ion exchange chromatography using DEAE- Cellulose in the presence of Tris buffer ( 0.05 Molar and pH 9 ). The obtained purification fold 25 and recovery 25% for A image which have specific activity 225 unit/ mg protein. The B image step obtained specific activity 375 unit/ mg protein , purification fold 41.6 and recovery 20.8 %. The optimum pH of activity for partially purified enzyme with ion exchange chromatography for both A and B image was 9 and 5 respectively and the optimum temperature was 35 C for both steps. | ||
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