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Production, Purification and Characterization of Tannase from Rhodococcus NCIM 2891

Author(s): Naiem H. Nadaf | Jai S. Ghosh

Journal: Current Research Journal of Biological Sciences
ISSN 2041-076X

Volume: 3;
Issue: 3;
Start page: 246;
Date: 2011;
Original page

Keywords: Gallic acid | NCIM 2891 | pollutants | Rhodococcus | tannase | tannin

The study was done with the objective of tannic acid degradation by tannase from the Rhodococcus NCIM 2891 which is actually a desulfurizing bacteria. Rhodococcus NCIM 2891cultivated in medium containing 0.1% Tannic acid, produced tannase which showed maximum activity after 24 h. The enzyme was purified by DEAE cellulose ion exchange chromatography and was shown to be dimeric in nature it has two subunits as Tannase I and Tannase II, having specific activities of 0.23 U/mg of protein and 0.295 U/mg of protein, respectively. Both the enzyme showed optimum activity at pH 6 and 30ÂșC. The Km of both enzymes was 0.034 and 0.040 mM respectively and Vmax of both enzymes were found to be 40 and 45 U/mL, respectively. SDS-PAGE analysis of purified proteins fraction revealed that their molecular weights are 60 and 62 kDa. Tannase I was completely inhibited by 10 mM Hg2+, Cu2+, Fe3+, Co2+ and Tannase I and II both was completely inhibited by1 mM Hg2+ alone. The DNA damage- protecting activity of tannic acid and product obtained after tannic acid hydrolysis was assessed bringing about the DNA damage with H2O2 + UV exposure in absence of the hydrolyzed products of tannic acid. The experiment was carried out by using agarose gel electrophoresis to analyze DNA damage in presence of H2O2 + UV. However, there was no damage of DNA by H2O2 + UV exposure in presence of the hydrolysis products.

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