Determination of Biochemical Properties of Polyphenol Oxidase from Domat Olives (In English)

Abstract

 Enzymatic browning of damaged tissues of fruits and vegetables occurs from the oxidation of phenolic compounds and contributes significantly to quality loss. The primary enzyme responsible for the browning reaction is polyphenol oxidase (PPO). PPO from Domat olives was extracted and purified through (NH4)2SO4 precipitation, dialysis and ion exchange chromatography. Optimum pH and temperature for olive PPO were found to be 4.5 and 30 °C, respectively, using 4-methyl catechol as substrate. Km and Vmax values for olive PPO using 4-methyl catechol as substrate were found to be 14.52 mM (r2= 0.985) and 1.78 Abs/min, respectively. Of the inhibitors tested, the most potent inhibitors were L-cysteine and sodium disulfite. The Z and Ea values obtained in this study were 18.6 °C (r2 = 0.953) and 121.0 kj/mol (r2= 0.950), respectively. 

Keywords

• Olive, polyphenol oxidase, enzymatic browning, kinetics, thermal inactivation

Domat Zeytini Polifenol Oksidazının Biyokimyasal Özelliklerinin Belirlenmesi (İngilizce)

Abstract

Zarar görmüş meye ve sebzelerdeki enzimatik esmerleşme fenolik bileşiklerin oksidasyonu sonucu meydana gelir ve önemli kalite kayıplarına neden olur. Esmerleşme reaksiyonuna neden olan en önemli enzim polifenol oksidaz (PFO)’dır. Domat zeytininden polifenol oksidaz enzimi ekstrakte edilmiş ve amonyum sülfat çökeltmesi, dializ ve iyon değişim kromatografisi ile saflaştırılmıştır. Zeytin PFO’sunun optimum pH ve sıcaklığı sırasıyla 4-metil kateşol substratı kullanılarak 4.5 ve 30 °C bulunmuştur. Zeytin PFO’su Km ve Vmax değerleri sırasıyla 4-metil kateşol substratı kullanılarak 14.52 mM (r2= 0.985) ve 1.78 Abs/dak bulunmuştur. Test edilmiş inhibitörlerin en etkili olanları sistein ve sodyum disülfittir. Bu çalışmada elde edilmiş Z ve Ea değerleri sırasıyla 18.6 °C (r2 = 0.953) ve 121.0 kj/mol (r2= 0.950) bulunmuştur.

Keywords

• Zeytin, polifenol oksidaz, enzimatik esmerleşme, kinetik, termal inaktivasyon

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