Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8359539 | Protein Expression and Purification | 2018 | 7 Pages |
Abstract
At first, commercial, crude tyrosinase from Agaricus bisporus (AbTyr) dissolved in aqueous media was added to octadecyl-Sepabeads matrix at 25â¯Â°C. Under these conditions, the support specifically adsorbed a protein with a molecular weight of 47â¯kDa which showed no tyrosinase activity. The known H subunit of tyrosinase from Agaricus bisporus (45â¯kDa, H-AbTyr) and another protein of 50â¯kDa were present in the supernatant. Sodium phosphate buffer was added to adjust the ionic strength of the solution up to 100â¯mM and Triton X-100 was added (final concentration of 0.07% v/v) to control the hydrophobicity effect for both proteins. This solution was offered again to fresh octadecyl-Sepabeads support, immobilizing selectively the H-AbTyr and leaving exclusively the 50â¯kDa protein as a pure sample in the supernatant. This tyrosinase isoform of 50â¯kDa was almost 4-fold more active than the known H-TyrAb, with a specific tyrosinase activity of more than 38,000 U/mg.
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Authors
David Lopez-Tejedor, Jose M. Palomo,