A New Approach to Prevent Enzymatic Browning in Sugar Beet: Herbiside Inhibitors

Öz

Enzymatic browning represents a significant challenge in the processing and storage of fresh produce within the food industry, as it can adversely affect consumer preferences. Various physical and chemical methods have been explored to inhibit the activity of polyphenol oxidase in order to extend the shelf life of products. In this study, it was aimed to prevent enzymatic browning in sugar beet (Beta vulgaris L. subsp. vulgaris) by limiting the polyphenol oxidase (PPO) activity with selected herbicides. Polyphenol oxidase enzyme was purified 46.35-fold by affinity chromatography using Sepharose-4B-L-tyrosine-p-aminobenzoic acid affinity gel. The molecular weight of the enzyme was found to be about 40 kDa by SDS-PAGE as a single band. Then, the inhibition effects of pendimethalin, bentazone, and glyphosate on purified PFO enzyme were investigated. IC50 values were determined as 0.157, 0.081, and 0.230 µM for pendimethalin, bentazone, and glyphosate herbicides, respectively. Kinetic assays indicated that bentazone was a remarkable inhibitor with the Ki value of 0.092 µM on PPO.

Anahtar Kelimeler

• • Polyphenol oxidase
• • purification
• • inhibition
• • herbicide

Şeker Pancarında Enzimatik Esmerleşmenin Önlenmesi İçin Yeni Bir Yaklaşım: Herbisit İnhibitörleri

Öz

Enzimatik esmerleşme, tüketici tercihlerini olumsuz etkileyebileceğinden, gıda endüstrisinde taze ürünlerin işlenmesi ve depolanmasında önemli bir zorluk teşkil etmektedir. Ürünlerin raf ömrünü uzatmak amacıyla, polifenoloksidaz enzim aktivitesini inhibe eden çeşitli fiziksel ve kimyasal yöntemler denenmiştir. Bu çalışmada, seçilmiş herbisitlerle polifenol oksidaz (PFO) aktivitesini sınırlayarak şeker pancarı (Beta vulgaris L. subsp. vulgaris) bitkisinde enzimatik esmerleşmenin önlenmesi amaçlanmıştır. Polifenol oksidaz enzimi, Sepharose-4B-L-tirozin-p-aminobenzoik asit afinite jeli kullanılarak afinite kromatografisi ile 46,35 kat saflaştırıldı. Enzimin molekül ağırlığı SDS-PAGE ile tek bant olarak yaklaşık 40 kDa olarak bulundu. Daha sonra, pendimetalin, bentazon ve glifosatın saflaştırılmış PFO enzimi üzerindeki inhibisyon etkileri araştırıldı. IC50 değerleri pendimetalin, bentazon ve glifosat herbisitleri için sırasıyla 0,157, 0,081 ve 0,230 µM olarak belirlendi. Kinetik analizler, bentazonun PFO üzerinde 0,092 µM'lik Ki değeriyle dikkate değer bir inhibitör olduğunu gösterdi.

Anahtar Kelimeler

• Polifenol oksidaz
• saflaştırma
• inhibisyon
• herbisit

Kaynakça

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