Beyaz Kiraz Meyvesi (Starks Gold) Polifenol Oksidazının Karakterizasyonu (İngilizce)

Beyaz kiraz meyvesinden (Starks Gold) polifenol oksidaz enzimi ekstrakte edilmiş ve amonyum sülfat çökeltmesi, diyaliz ve iyon değişim kromatografisi ile saflaştırılmıştır. Toyopearl 650 m kolon kromatografisi sonucunda PFO aktivitesi gösteren izoenzim a ve izoenzim b ile belirtilmiş iki pik elde edilmiştir. İzoenzim a % 34.9 verimle 3.9 kat, izoenzim b % 54.3 verimle 76.7 kat saflaştırılmıştır. İzoenzim a ve b’nin optimum pH değerleri sırası ile 4.5 ve 4.98 olarak bulunmuştur. Optimum sıcaklık değerinin izoenzim a için 20°C ve izoenzim b için 30°C olduğu saptanmıştır. İzoenzim b’nin kateşol substratına ilgisi izoenzim b’den daha yüksektir. Aktivasyon enerjisi ve z değerleri izoenzim a için 22.1°C (r2= 0.883) ve 98.5 kj/mol (r2= 0.878), izoenzim b için 13.9°C (r2= 0.990) ve 157.1 kj/mol (r2= 0.989) bulunmuştur. L-sistein ve sodyum disülfitin inhibitör etkisinin birbirinden farklı olduğu belirlenmiştir.

Anahtar Kelimeler:

Beyaz kiraz, polifenol oksidaz, enzimatik esmerleşme, kinetik, termal inaktivasyon

Characterization of Polyphenol Oxidase From White Cherry Fruit (Starks Gold) (in English)

Polyphenol oxidase (PPO) from white cherry fruit (Starks Gold) was extracted and purified through (NH4)2SO4 precipitation, dialysis and ion exchange chromatography. The enzyme showed two peaks with PPO activity on toyopearl 650 m column, which were denoted as isoenzyme a and isoenzyme b. A 3.9 fold purification of isoenzyme a with a recovery of 34.9 % and 76.7 fold purification of isoenzyme b with a recovery of 54.3 % were achieved. Ph optima of isoenzyme a and b were 4.5 and 4.98, respectively. The temperature optima for enzyme activity were found to be 20°C for isoenzyme a and 30°C for isoenzyme b. The affinity of isoenzyme b for catechol as substrate was higher than that of isoenzyme a. Activation energies and z values were found to be 22.1°C (r2= 0.883) and 98.5 kj/mol (r2= 0.878) for isoenzyme a and 13.9°C (r2= 0.990) and 157.1 kj/mol (r2= 0.989) for isoenzyme b, respectively. Inhibitory effect of l-cysteine and sodium disulphide differed from each other.

Keywords:

White cherry, polyphenol oxidase, enzymatic browning, kinetics, thermal inactivation,

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