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KMID : 0545120100200101403
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Journal of Microbiology and Biotechnology
2010 Volume.20 No. 10 p.1403 ~ p.1414
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Acidophilic tannase from marine Aspergillus awamori BTMFW032
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P. S. Beena
M. B. Soorej
K. K. Elyas
G. Bhat Sarita
M. Chandrasekaran
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Abstract
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Aspergillus awamori BTMFW032, isolated from sea water, produced tannase as extracellular enzyme under submerged culture conditions. Enzyme with a specific activity of 2761.89 IU/mg protein, a final yield of 0.51 %, and a purification fold of 6.32 was obtained after purification to homogeneity by ultrafiltration and gel filtration. SDS-PAGE analyses under non- reducing and reducing conditions yielded a single band of 230 kDa and 37.8 kDa, respectively, indicating presence of six identical monomers. pI of 4.4 and 8.02 % carbohydrate content in the enzyme were observed. Optimal temperature was 30ºC, although the enzyme was active at 5-80 ºC. Two pH optima, pH 2 and pH 8, were recorded and the enzyme was stable only at pH 2.0 for 24 h. Methylgallate recorded maximal affinity and Km and Vmax were recorded, respectively, as 1.9 X 10-3 M and 830 μmol/min.. Impact of several metal salts, solvents, surfactants, and typical enzyme inhibitors on tannase activity were determined to establish the novelty of the enzyme. Gene encoding tannase isolated from A. awamori is 1.232 kb and nucleic acid sequence analysis revealed an open reading frame consisting of 1122 bp (374 amino acids) of one stretch in -1 strand. In-silico analyses of gene sequences and comparison with reported sequences of other species of Aspergillus indicated that the acidophilic tannase from marine A. awamori is different from others.
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KEYWORD
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Aspergillus awamori, acidophilic tannase, characterization
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