Purification and characterization of laccase from Ganoderma sp. 03
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Abstract
Laccases are enzymes that catalyze oxidation of a wide range of substrates. The enzyme shows promising applications in various fields such as toxic substance degradation, organic synthesis, and biosensors. In this study, we purified and characterized laccase from Ganoderma sp. Three different Ganoderma sp. found in the northeast Thailand were studied for laccase activity. When cultured in media containing rice bran and rice husk, Ganoderma sp. 03 showed high laccase activity. The laccase produced by this strain was purified using Phenyl Sepharose fast flow (FF) chromatography and quaternary amine (Q) Sepharose chromatography. Estimated molecular weight of the purified laccase was 39.81 kDa as determined by the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The purified laccase had an optimal potential hydrogen (pH) of 3.5, showed high stability at pH 3.0-5.0 using 2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) as a substrate, and exhibited an optimal temperature of 40.0-55.0°C. The best temperature stability was observed at 30°C. Effect of different substances on laccase activity showed that p-coumaric acid and dithiothreitol (DTT) strongly inhibited laccase activity. Especially, DTT could inhibit 100% of laccase activity, even at a concentration as low as 0.1 mM. All the tested metal ions, except ZnSO4, decreased laccase activity. This information may be useful for further studies and applications of this enzyme.
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