Production, Purification and Characterization of Tannase from Rhodococcus NCIM 2891

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 30°C. The K_m of both enzymes was 0.034 and 0.040 mM respectively and V_(max) 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 Hg^(2+), Cu^(2+), Fe^(3+), Co^(2+) and Tannase I and II both was completely inhibited by1 mM Hg^(2+) alone. The DNA damage- protecting activity of tannic acid and product obtained after tannic acid hydrolysis was assessed bringing about the DNA damage with H_2O_2 + 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 H_2O_2 + UV. However, there was no damage of DNA by H_2O_2 + UV exposure in presence of the hydrolysis products.

並列關鍵字

Gallic acid ； NCIM 2891 ； pollutants ； Rhodococcus ； tannase ； tannin

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