A cDNA Cloning of a Novel Alpha-Class Tyrosinase of Pinctada fucata: Its Expression Analysis and Characterization of the Expressed Protein

Figure 5

Measurement of the enzyme activity of the in vitro translation product and mushroom tyrosinase. The spectrophotometric tyrosinase assay at 475 nm was performed as described in Section 2. (a) In the absence of EDTA; the translation product of pET29b-Tlg-cDNA (◆-◆), the translation product of pET29b (■-■), blank (●-●). (b) Mushroom tyrosinase (○-○), the translation product of pET29b-Tlg-cDNA (◆-◆), mushroom tyrosinase + EDTA (△-△), the translation product of pET29b-Tlg-cDNA + EDTA (▲-▲), the translation product of pET29b + EDTA (■-■), blank + EDTA (●-●). Blank is a 500 μL solution of the substrate (L-DOPA). Abbreviations are as follows: Tlg-cDNA, tyrosinase-like gene cDNA. The used amount of mushroom tyrosinase was 0.1 unit (0.1 μg) dissolved in the 20 μL of H₂O.

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(b)