Purification of tyrosinase from edible mushroom

A simple preparative method was developed for purification of Tyrosinase from edible mushroom (Agaricus bispora). A homogenized extract of mushroom was first saturated by ammonium sulfate. The desired precipitate was mixed thoroughly with DEAE-Cellulose (DE-52) and washed out to produce melanin free precipitate. The obtained protein solution was dialyzed against running water for 4 hrs, then, concentrated and chromatographed on a DE-52 column. On the basis of the activities assay, the eluted fractions by 150 mM salt solution were selected for further purification. The collected fractions were pooled and chromatographed on a Sephadex G-200 column. Polyacrylamide gel electrophoresis (PAGE) of the purified tyrosinase produced a single band right beside the commercial sample obtained from Sigma Company at 128 kDa. The lyophilized form of the purified Tyrosinase had a purification degree of 104 and showed strong cresolase and catecholase activities when compared to a commerically available tyrosinase.

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