Abstract
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Article Information:
Production, Purification and Characterization of Tannase from Rhodococcus NCIM 2891
Naiem H. Nadaf and Jai S. Ghosh
Corresponding Author: Jai S. Ghosh
Submitted: 2011 March, 24
Accepted: 2011 April, 20
Published: 2011 May, 05 |
Abstract:
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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.
Key words: Gallic acid, NCIM 2891, pollutants, Rhodococcus, tannase, tannin,
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Cite this Reference:
Naiem H. Nadaf and Jai S. Ghosh, . Production, Purification and Characterization of Tannase from Rhodococcus NCIM 2891. Current Research Journal of Biological Sciences, (3): 246-253.
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ISSN (Online): 2041-0778
ISSN (Print): 2041-076X |
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