Purification and properties of tyrosinases from Vibrio tyrosinaticus1

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Two tyrosinase isoenzymes (EC1. 10.3.1) have been highly purified from Vibrio tyrosinaticus. This represents the first characterization of tyrosinases from eubacteria. The enzymes have molecular weights of 41,000 and 38,500, closer to the tyrosinase of Neurospora crassa (33,000) than to the mammalian enzymes from mouse and hamster melanoma (55,000 and 70,000). The bacterial tyrosinases are also different from the mammalian enzymes in that the former possess basic isoelectric points. The Vibrio tyrosinases do not cross-react with antiserum against a hamster melanoma tyrosinase. The Km values for l-tyrosine are 3.1 mm for both isoenzymes while the Km value for l-DOPA is 36 mm for one and 67 mm for the other. The values for tyrosine are tenfold, and those for dopa about 100-fold, larger than the corresponding Km values for the tumor enzymes. Apoenzyme was prepared by treatment with diethyldithiocarbamate, and activity was partially or wholly restored by addition of Cu2+, Mn3+, Cd2+, or Fe2+. The one isoenzyme examined has some activity on d-tyrosine and very slight activity against d, l-m-tyrosine. No activity was detected with catechol or l-phenylalanine.

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    This research was supported by Research Grant CA-07093 from the National Cancer Institute, National Institutes of Health. A portion of this work is derived from a Dissertation presented by V. V. Murthy to the Graduate School of the University of Maryland in 1968 in partial fulfillment of the Ph.D. degree.

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