Immobilzation of polyphenol oxidase enzyme with different matters and investigation of the some properties

The purpose of this study was the characterization of Polyphenol oxidase (PPO) enzyme from rocket (Eruca vesicaria) in the Sakarya region and immobilization of this isolated enzyme with different immobilization chemicals, Ca-alginate, Ba-alginate, polyacrilamide and alginate-polyacrylamide and the investigation of some properties of free and immobilized rocket PPO enzyme. Thus, optimum pH and temperatures of rocket PPO were determined for five different substrates (chatechole, 4-methylcathecol, pyrogallol, L-Tyrosine and gallic acid). The best substrate for rocket PPO was 4-methylchatecole with min Km value. Optimum pH and temperature of rocket PPO were determined 5.0 and 20 oC for 4-methylcathecole. Kinetic parameters of 4-methylchatecole were calculated 3,68 mM for Km and 16,6.10-3 EU/min for Vmax for free rocket PPO. Then, the characterized PPO enzyme was successfully immobilized with Ca-alginate, Ba-alginate, polyacrilamide and alginate-polyacrylamide and investigated kinetically and compared with the free enzyme. Kinetic parameters were calculated as 4,33mM , 1,45mM , 2,41 mM, 2,93 mM, Vmax değerleri ise 3,2. 10-3 EU/min, 4,0 10-3 EU/min, 9,2.10-3 EU/min, 10,0 10-3 EU/min for Ca-alginate, Ba-alginate, alginate-polyacrilamide and polyacrylamide immobilized enzyme. The optimum pH and temperature were parallel for free ad immobilized enzyme. The Immobilized enzyme was very stable compare to free enzyme in the same conditions at room temperature. The immobilized PPO was retained 26 % of the initial activity was used repeatedly 12 times in two days, respectively.Key Words: Polyphenol oxidase, rocket, Eruca vesicaria, immobilization of enzyme, alginate, polyacrylamide