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Structural characterization of thermostable, solvent tolerant, cytosafe tannase from Bacillus subtilis PAB2
•Tamarind seed can be successfully used as cheap substrate for tannase production.•Bacillus subtilis PAB2 yields a 52kDa tannase with globular secondary structure.•Tannase exhibits stability at a wide range of temperature, salt and organic solvent.•Native conformation of tannase alike a spherical na...
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Published in: | Biochemical engineering journal 2013-08, Vol.77, p.161-170 |
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Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
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Online Access: | Get full text |
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Summary: | •Tamarind seed can be successfully used as cheap substrate for tannase production.•Bacillus subtilis PAB2 yields a 52kDa tannase with globular secondary structure.•Tannase exhibits stability at a wide range of temperature, salt and organic solvent.•Native conformation of tannase alike a spherical nanoparticle.•Tannase have no cytotoxicity on Vero cell line and rat model.
Tannase production by Bacillus subtilis PAB2, was investigated under solid state fermentation using tamarind seed as sole carbon source and it was found as the highest titer (73.44U/gds). The enzyme was purified to homogeneity, which showed the molecular mass around 52kDa (Km=0.445mM, Vmax=125.8mM/mg/min and Kcat=2.88min–1). The enzyme was found stable in a range of pH (3.0–8.0) and temperature (30–70°C) with an optimal activity at pH 5.0, pI of 4.4 and at 40°C temperature. It exhibited half-life (t1/2) of 4.5h at 60°C. The enzyme comprised a typical secondary structure containing α-helix (9.3%), β-pleated sheet (33.6%) and β-turn (17.2%). The native conformation of the enzyme was alike a 44nm spherical nanoparticle upon aggregation. Thermodynamic parameters of tannase revealed that it was stable at 40°C and showed Q10, ΔGd and ΔSd values of 2.08, 99.37KJ/mol and 252.38Jmol−1K−1, respectively. Organic solvents were stimulatory with regard to enzyme activity. Moreover, the altered enzyme activity was determined to be correlated with the changes in structural conformation in presence of inducer and inhibitor. Tannase was explored to have no cytotoxicity on Vero cell line as well as rat model study. |
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ISSN: | 1369-703X 1873-295X |
DOI: | 10.1016/j.bej.2013.06.002 |