Abstract
A laccase from the culture filtrate of Phellinus linteus MTCC-1175 has been purified to homogeneity. The method involved concentration of the culture filtrate by ammonium sulphate precipitation and an anion exchange chromatography on DEAE-cellulose. The SDS-PAGE and native-PAGE gave single protein band indicating that the enzyme preparation was pure. The molecular mass of the enzyme determined from SDS-PAGE analysis was 70 kDa. Using 2.6-dimethoxyphenol, 2.2′[azino-bis-(3-ethylbonzthiazoline-6-sulphonic acid) diammonium salt] (ABTS) and 4-hydroxy-3,5-dimethoxybenzaldehyde azine as the substrates, the K m, k cat and k cat/K m values of the laccase were found to be 160 μM, 6.85 s−1, 4.28 × 104 M−1 s−1, 42 μM, 6.85 s−1, 16.3 × 104 M−1 s−1 and 92 μM, 6.85 s−1, 7.44 × 104 M−1 s−1, respectively. The pH and the temperature optima of the P. linteus MTCC-1175 laccase were 5.0 and 45°C, respectively. The activation energy for thermal denaturation of the enzyme was 38.20 kJ/mole/K. The enzyme was the most stable at pH 5.0 after 1 h reaction. In the presence of ABTS as the mediator, the enzyme transformed toluene, 3-nitrotoluene and 4-chlorotoluene to benzaldehyde, 3-nitrobenzaldehyde and 4-chlorobenzaldehyde, respectively.
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Chaurasia, P.K., Yadav, A., Yadav, R.S.S. et al. Purification and characterization of laccase secreted by Phellinus linteus MTCC-1175 and its role in the selective oxidation of aromatic methyl group. Appl Biochem Microbiol 49, 592–599 (2013). https://doi.org/10.1134/S000368381306006
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DOI: https://doi.org/10.1134/S000368381306006