ISOLATION OF CELLULOLYTIC ENZYMES FROM THE FUNGUS TRICHODERMA HARZIANUM В1
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Authors
B.R. Umarov
Institute of Microbiology, Academy of Sciences of Uzbekistan, 7b, A. Kadyri str., Tashkent, 100128, Uzbekistan
A.V. Sagdiev
A.S. Sadykov Institute of Bioorganic Chemistry, Academy of Sciences of the Republic of Uzbekistan, 83, M. Ulugbek str., Tashkent, 100125, Uzbekistan
A.V. Kim
A.S. Sadykov Institute of Bioorganic Chemistry, Academy of Sciences of the Republic of Uzbekistan, 83, M. Ulugbek str., Tashkent, 100125, Uzbekistan
U.K. Inagamov
A.S. Sadykov Institute of Bioorganic Chemistry, Academy of Sciences of the Republic of Uzbekistan, 83, M. Ulugbek str., Tashkent, 100125, Uzbekistan
Abstract
The local strain of Trichoderma harzianum В1 produces cellulolytic enzymes when cultured submerged in a medium containing 2% wheat bran as the sole carbon source. Chromatography systems using different carriers and different matrices, including Sephadex G-75, P-60 Akrileks, and ion exchange chromatography on DEAE 650M gel Toyperl, were used to isolate three isoforms of endo-1,4-β-glucanase (EC 3.2.1.4), EG 1, EG 2, and EG 3 and determined a molecular weight (MW) of 35±1 kDa. The isolates exhibited a cellulolytic activity of 90.4, 77.52, and 78.92 U/mg protein. In addition, we isolated four isoforms of cellobiase (1.3-β-glucosidase, EC 3.2.1.21) CBH 1, CBH 2, CBH 3, and CBH 4 with aMW 24±1 kDa, and a cellulolytic activity of 2.60, 3.80, 4.3, and 3.0 U/mg protein. The influence of the temperature, pH, and metal ions on the activity was determined for the cellulolytic enzymes. The optimal pH and temperature of endo-1,4-β-glucanase and cellobiase (1,3-β-glucosidase) was observed to bepH 4.8 and 50°C. The effect of the monovalent and divalent metal ions K, Na, Fe, Mg, and Mn in a buffered medium increased the enzymatic activity of 1,4-β-glucanase by10%, and of 1,3-β-glucosidase by 15%.
Keywords
Trichoderma harzianum В1, cellulolytic enzymes, 1,3-β-glucosidase, endo-1,4-β-glucanase
Article Details
References
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