Purification and characterization of the tyrosinase isozymes of pine needles.

Three tyrosinase isozymes were purified to electrophoretic homogeneity from pine needles. The molecular weight of the three isozymes (P1, P2 and P3) were approximately 65,000, 50,000 and 45,000, and the pI values were 6.2, 5.9 and 5.3, respectively. The three isozymes have a number of common properties. These include amino acid composition, substrate specificity, response to inhibition. The amino acid compositions of the three isozymes showed the characteristic high contents of glycine, serine and glutamic acid residues. The three isozymes exhibited high substrate specificity towards pyrogallol. The K(m) values of the three isozymes for L-DOPA ranged from 8.7 to 10 mM. L-ascorbic acid and beta -mercaptoethanol, glutathione and sodium diethyldithiocarbamate notably inhibited the enzymatic activities.