Extraction, purification of tyrosinase and using removal of phenol

In this study, tyrosinase was extracted and purified from green grape. This extracted and purified tyrosinase was used for separation of phenol from aqueous solutions containing phenol. Phenol separation was also studied with pure commercial tyrosinase. Phenol separation efficiencies observed with purified enzyme and commercial enzyme were compared. Extraction and purification steps are firstly crude enzyme extraction, then purification with Triton X-100 and PVP (polyvinyl pyrrolidone) and finally precipitation with ammonium sulphate. The effects of Triton X-100 and PVP (polyvinyl pyrrolidone) concentrations on tyrosinase spesific activity were investigated. The maximum spesific activity of tyrosinase (2874,51 U/mg protein) was observed with 3 % (w/v) PVP (polyvinyl pyrrolidone) and 4,5 % (v/v) Triton X-100 concentrations in the experimantal range studied. The effects of enzyme concentration, separation time and temperature on phenol removal efficiency were investigated. The maximum phenol removal efficiency was achieved at 25 oC and with a separation time of three hours. Phenol separation efficiencies obtained with purified enzyme and commercial enzyme were very close each other. Using equal enzyme activities, the phenol separation efficiencies were found as 75-79 % and 92-95 % with purified tyrosinase enzyme and commercial enzyme, respectively. Increasing the enzyme activity by 50 % for the separation experiments with the purified tyrosinase enzyme in this work resulted in an increase in the phenol separation efficiency to 82-89 %.