Comparison of Polyphenol Oxidase Enzymes in Fresh and Dried Agaricus bisporus

Öz

Agaricus bisporus (J.E. Lange) Imbach is the most important mushroom species cultivated commercially in our country as well as in the world. The polyphenol oxidase (PPO) is an enzyme that catalyzes the oxidation of phenolic compounds commonly found in fruits and vegetables, oxidizes them to o-quinones, and leads to the formation of brown melanin pigments that cause browning as a result of their polymerization. Enzymatic browning reactions catalyzed by the PPO enzyme are undesirable because they reduce the taste, appearance, and nutritional value of the product. In this study, it was aimed to compare the PPO enzyme of fresh and dried A. bisporus. Extraction, purification (ammonium sulfate precipitation, dialysis, and affinity column chromatography), and characterization of PPO enzymes present in fresh and dry A. bisporus were performed. Sepharose 4B-L-tyrosine-tyrosine-p-aminobenzoic acid was used as an affinity gel for purification. PPO activities in purified fresh and dried A. bisporus were evaluated using the spectrophotometric method. Catechol was used as a substrate for the determination of PPO enzyme activities. As a result of the study, it was determined that the PPO activities of fresh and dried A. bisporus were different. This is the first comparative study on PPO enzymes in fresh and dried A. bisporus.

Anahtar Kelimeler

• Affinity Chromatography
• Characterization
• Agaricus bisporus
• Polyphenol Oxidase

Taze ve Kuru Agaricus bisporus’da Polifenol Oksidaz Enzimlerinin Karşılaştırılması

Öz

Agaricus bisporus (J.E. Lange ) Imbach, dünyada olduğu gibi ülkemizde de ticari olarak kültürü yapılan en önemli mantar türlerinden biridir. Polifenol oksidaz (PPO), meyve ve sebzelerde yaygın olarak bulunan fenolik bileşiklerin oksidasyonunu katalizleyerek, onları o-kinonlara yükseltgeyen ve bunların polimerizasyonu sonucu esmerleşmeyi yapan kahverengi melanin pigmentlerinin oluşumuna yol açan bir enzimdir. PPO enziminin katalizlediği enzimatik kararma reaksiyonları ürünün tat, görünüm ve besin değerini düşürdüğünden istenmemektedir. Bu çalışmada, A. bisporus’un taze ve kuru halinin PPO enziminin karşılaştırılması amaçlanmıştır. Taze ve kuru A. bisporus’da mevcut olan PPO enzimlerinin ekstraksiyonu, saflaştırılması (amonyum sülfat çöktürmesi, diyaliz ve afinite kolon kromatografisi) ve karakterizasyonu yapılmıştır. Saflaştırmada afinite jeli olarak Sepharose 4B-L-tirozin-p-aminobenzoik asit kullanılmıştır. Saflaştırılan taze ve kuru A. bisporus’daki PPO aktiviteleri spektrofotometrik yöntem kullanılarak değerlendirilmiştir. PPO enzim aktivitelerinin belirlenmesi için substrat olarak katekol kullanılmıştır. Çalışma sonucunda, A. bisporus’un taze ve kuru hallerinin PPO aktivitelerinin farklı olduğu tespit edilmiştir. Bu araştırma, taze ve kuru A. bisporus’daki PPO enzimlerinin karşılaştırılmalı olarak incelendiği ilk çalışmadır.

Anahtar Kelimeler

• Afinite Kromatografisi
• Karakterizasyon
• Agaricus bisporus
• Polifenol Oksidaz

Kaynakça

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