1. A modification of the Nelson and Mason (1970) method for tyrosinase (EC 1.14.18.1) extraction, purification and separation from Agaricus bisporus is proposed. 2. Modifications primarily concern: (a) enzyme recovery by performing pellet re-extractions, (b) preparative isoelectric focusing purification replacing a gel-permeation, (c) use of polyvinyipyrrolidone to remove contaminant ''brown proteins'', instead of basic Pb acetate. 3. With this procedure, it is possible to enhance the amount of tyrosinase recovered, thus reducing the quantity of mushrooms required (90% less). The enzyme obtained displays a better degree of purification (between 81- and 92-fold), and the total extraction time is lower than that of the original technique.
THE AGARICUS BISPORUS TYROSINASE 1: PROGRESS MADE IN PREPARATIVE METHODS.
PESSIONE, Enrica;GIUNTA, Carlo
1994-01-01
Abstract
1. A modification of the Nelson and Mason (1970) method for tyrosinase (EC 1.14.18.1) extraction, purification and separation from Agaricus bisporus is proposed. 2. Modifications primarily concern: (a) enzyme recovery by performing pellet re-extractions, (b) preparative isoelectric focusing purification replacing a gel-permeation, (c) use of polyvinyipyrrolidone to remove contaminant ''brown proteins'', instead of basic Pb acetate. 3. With this procedure, it is possible to enhance the amount of tyrosinase recovered, thus reducing the quantity of mushrooms required (90% less). The enzyme obtained displays a better degree of purification (between 81- and 92-fold), and the total extraction time is lower than that of the original technique.
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