Department of Biochemistry, National Research Institute for Genetic Engineering and Biotechnology, Tehran, I.R. Iran.
Abstract
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.
Haghbeen, K., Rastgar Jazii, F., Karkhane, A. A., & Shareefi Borojerdi, S. (2004). Purification of tyrosinase from edible mushroom. Iranian Journal of Biotechnology, 2(3), 189-194.
MLA
Kamahldin Haghbeen; Ferdous Rastgar Jazii; Ali Asghar Karkhane; Shahrzad Shareefi Borojerdi. "Purification of tyrosinase from edible mushroom". Iranian Journal of Biotechnology, 2, 3, 2004, 189-194.
HARVARD
Haghbeen, K., Rastgar Jazii, F., Karkhane, A. A., Shareefi Borojerdi, S. (2004). 'Purification of tyrosinase from edible mushroom', Iranian Journal of Biotechnology, 2(3), pp. 189-194.
VANCOUVER
Haghbeen, K., Rastgar Jazii, F., Karkhane, A. A., Shareefi Borojerdi, S. Purification of tyrosinase from edible mushroom. Iranian Journal of Biotechnology, 2004; 2(3): 189-194.