Bazı makrohalkalı tiyocrown eterlerin polifenol oksidaz enzimi üzerindeki inhibisyon etkilerinin araştırılması

Abstract

Polifenol oksidaz (PPO) enzimi, meyvelerde, sebzelerde ve bazı hayvansal dokularda bolca bulunabilen, aktif bölgesinde bakır olan bir metalo enzimdir.  Enzimatik esmerleşme reaksiyonlarını katalizleyen bu enzimin, ticari değere sahip ürünlerde bolca bulunmasından dolayı inhibisyonu oldukça önem kazanmıştır.  Bunun için literatürde birçok çalışma mevcuttur.  Bu çalışmada, PPO enzimi, amonyum sülfat çöktürmesi ve diyaliz gibi ön saflaştırma işlemlerinden sonra, Sepharose 4B-l-tirozin -p-aminobenzoik asit afinite kromatografi jeli kullanılarak, Musa sapientum var. Cavendishii (Muz) meyvesinden saflaştırılmıştır.  Enzimin saflık kontrolü de SDS – PAGE ile yapılmıştır.  Daha sonra, bizim tarafımızdan esterleşme – halka kapama reaksiyonu sonucunda sentezlenen AE1, AE2 ve AE3 kodlu makrohalkalı tiyocrown eterlerin, saf olarak elde ettiğimiz polifenoloksidaz enzimi üzerindeki inhibisyon etkileri araştırılmıştır.  Tüm maddelerin PPO enzimini inhibe ettiği saptanmıştır.  Sonuçlara göre en etkili inhibitör AE3 olarak tespit edilmiştir. 

Keywords

• Tiyocrown eter,polifenol oksidaz (PPO),inhibisyon,Afinite kromatografisi

Investigation of the ınhibitory effects of some macrogenetic thiocrown ethers on polyphenol oxidase enzyme

Abstract

The polyphenol oxidase (PPO) enzyme is a metallo enzyme that is copper in its active site, which can be found abundantly in fruits, vegetables and some animal tissues. Inhibition of this enzyme, which catalyzes the enzymatic browning reactions, is gaining in importance due to its abundance in commercial value products.  There are many studies in the literature for this.  In this study, PPO enzyme was purified using a Sepharose 4B-1-tyrosine-p-aminobenzoic acid affinity chromatography gel, after preliminary purification, such as ammonium sulfate precipitation and dialysis, from Musa sapientum var. Cavendishii (Banana).  Purity control of the enzyme was also performed by SDS - PAGE. We then investigated the inhibitory effects of the macrophilic thiocrown ethers AE1, AE2 and AE3 synthesized as a result of the esterification-ring closure reaction on the pure polyphenoloxidase enzyme.  All substances were found to inhibit PPO enzyme.  According to the results, the most effective inhibitor was identified as AE3.

Keywords

• Thiocrown ether,polyphenol oxidase (PPO),inhibition,affinity chromatography

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