Prevention of Enzymatic Browning by Inhibiting Polyphenol Oxidase with Some Natural Compounds and Benzenethiol

Abstract

Background: In recent years consumers' awareness of the health benefits of fresh vegetables and fruits have increased and demand for ready-made food has increased. That's why, fresh cut vegetables and fruits have become a popular. An important concern in this process is enzymatic browning, which cause changes in the texture, taste, and color of freshly cut product. Physical and chemical-based anti-browning methods to prolong the shelf life of freshly cut products are remarkable. In this study, it was aimed to prevent enzymatic browning via inhibition of polyphenol oxidase (PPO) enzyme. Method: Firstly, potato was used as the source of polyphenol oxidase and enzyme purified by affinity chromatography. Sepharose 4B L-tyrosine p-amino benzoic acid (Sepharose 4B L-tyr-p-ABA) was used as affinity column. SDS-PAGE was used to check enzyme purity. Then, the inhibition effect of baicalin, bailcalein, phloridzin, phloretin natural compounds and benzenthiol sulfur compound were investigated. Results: PPO was purified with 13.94% fold from potato. In inhibition studies Ki values of phloridzin, phloretin and benzenethiol were determined as 0.120±0.0157, 0.027±0.0054, 0.008±0.0014 mM respectively. According to our results, the order of inhibition is as follows: pholoridzin ˃ phloretin ˃ benzenethiol. While baicalin and bailcalein did not show any inhibition effect, phloridzin and phloretin showed non-competitive, benzentiol showed competitive inhibition.

Keywords

• Enzymatic Browning
• Inhibition
• Polyphenol Oxidase
• Natural Compound
• Benzenethiol

Bazı Doğal Bileşikler ve Benzentiyol ile Polifenol Oksidaz İnhibisyonu Yoluyla Enzimatik Kararmanın Önlenmesi

Öz

Arkaplan: Taze kesilmiş sebzeler ve meyveler, son yıllarda tüketicilerin taze sebzelerin ve meyvelerin sağlık yararları konusundaki farkındalığının artması ve hazır gıda talebinin artması nedeniyle popüler hale gelmiştir. Önemli bir endişe, taze kesilmiş ürünlerin renk, tat ve dokusunda değişikliklere yol açabilen ve bu nedenle kalite bozulması ile ilişkili enzimatik esmerleşmedir. Taze kesilmiş ürünlerin raf ömrünü uzatmak için fiziksel ve kimyasal temelli anti-esmerleşme yöntemleri dikkat çekicidir. Bu çalışmada, enzimatik esmerleşmeden sorumlu polifenol oksidaz (PPO) enzimi için inhibitörler önerilmiştir. Yöntem: İlk olarak patates, Sepharose 4B L-tirozin p-amino benzoik asit (Sepharose 4B L-tyr-p-ABA) afinite kromatografisi ile polifenol oksidaz saflaştırılmasında enzim kaynağı olarak kullanıldı. Enzim saflığı SDS-PAGE ile kontrol edildi. Daha sonra, baicalin, bailcalein, phloridzin, phloretin doğal bileşikleri ve benzentiol sülfür bileşiğinin inhibisyon etkisi araştırıldı. Sonuçlar: PPO, patatesden % 13.94 kat saflaştırıldı. İnhibisyon çalışmalarında phloridzin, phloretin ve benzenethiol'ün Ki değerleri sırasıyla 0.120 ± 0.0157, 0.027 ± 0.0054, 0.008 ± 0.0014 mM olarak belirlenmiştir. Sonuçlarımıza göre, inhibisyon sırası aşağıdaki gibidir: pholoridzin ˃ phloretin ˃ benzenethiol. Baicalin ve bailcalein bir inhibisyon etkisi göstermezken, phloridzin, phloretin yarşmasız, benzentiol yarışmalı inhibisyon gösterdi.

Anahtar Kelimeler

• Enzimatik Kararma
• İnhibisyon
• Polifenol Oksidaz
• Doğal Bileşikler
• Benzentiol

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