Sonesson B, Eide S, Rorsman H, Rosengren E
DOI: 10.2340/0001555575283286

Serum samples for the analysis of tyrosinase activity were obtained from 10 healthy subjects in autumn, winter and summer. Tyrosinase was purified from 100 microl serum by adsorption to concanavalin A sepharose, the tyrosinase adsorbed to the gel being separated from other components by centrifugation. The gel was suspended in a buffer containing 5-hydroxy-indole-3-acetic acid as an antioxidant and incubated for 2 min with L-cysteine and D-L-dopa at 37 degrees C. The 5-S-L-cysteinyl-L-dopa formed was measured by HPLC and electrochemical detection. Tyrosinase has high stereo-specificity for the L-enantiomer of dopa, and correction for non-specific oxidation was made by simultaneous measurement of 5-S-L-cysteinyl-D-dopa formed from D-dopa. Whereas the oxidation of L-dopa catalysed by tyrosinase ws inhibited by L-tyrosine, the non-specific oxidation of D-dopa was not. Mean serum tyrosinase activity was 0.9 nkatal/l in summer, 0.8 nkatal/l in autumn and 0.4 nkatal/l in winter. The range of tyrosinase activity was much higher in summer and autumn than in winter