Department of Food Engineering, Faculty of Engineering, Iğdır University, Iğdır, Turkey.
J Food Biochem. 2021 Feb;45(2):e13627. doi: 10.1111/jfbc.13627. Epub 2021 Feb 1.
The purification of the polyphenol oxidase (PPO) enzyme from Kirmizi Kismis grape (Vitis vinifera L.) was performed 61.23 times using affinity chromatography. The molecular weight of the enzyme was found to be about 38.1 kDa by SDS-PAGE as a single band. The optimum pH and temperature values were revealed to be 5.0 and 30°C, respectively, in the presence of 4-methyl catechol substrate. The thermal stability of PPO was examined and it was observed to maintain its activity at 20°C for 1 hr. K and V values were determined to be 4.8 mM and 2000,0 EU/ml for 4-methyl catechol as a substrate. IC and K values and inhibition types were found for various browning agents and ascorbic acid had the strongest inhibitory impact on PPO. The inhibitory impact of Na , K , Mg , Cu , and Al metal ions on the enzyme activity at final concentrations of 1 mM and 10 mM was examined. PRACTICAL APPLICATIONS: Grapes grown and processed take a significant place in our life. The grape has antioxidant, anticarcinogenic, antidiabetic and protective properties against bacteria and viruses. Furthermore, it takes an important position in the country's economy and social life due to providing raw materials to the food industry and having high export potential. Polyphenol oxidase, which is the leading actor of enzymatic browning reactions causing serious economic losses every year, was purified and characterized from Kirmizi Kismis grape (Vitis vinifera L.). This ancient grape variety has industrial processing and export potential due to its long storage life and resistance to oxidation. Therefore, the purification and biochemical characterization of polyphenol oxidase from Kirmizi Kismis grape are of great importance.
从红玫瑰葡萄（Vitis vinifera L.）中通过亲和层析对多酚氧化酶（PPO）进行了 61.23 倍的纯化。SDS-PAGE 显示酶的分子量约为 38.1 kDa，为单一条带。在 4-甲基儿茶酚底物存在下，发现最适 pH 和温度值分别为 5.0 和 30°C。研究了 PPO 的热稳定性，发现其在 20°C 下保持 1 小时的活性。确定 K 和 V 值分别为 4.8 mM 和 2000,0 EU/ml，4-甲基儿茶酚为底物。对于各种褐变剂，测定了 IC 和 K 值以及抑制类型，并且抗坏血酸对 PPO 具有最强的抑制作用。在终浓度为 1 mM 和 10 mM 时，研究了 Na 、K 、Mg 、Cu 和 Al 金属离子对酶活性的抑制作用。
生长和加工的葡萄在我们的生活中占有重要地位。葡萄具有抗氧化、抗癌、抗糖尿病和保护作用，可以抵抗细菌和病毒。此外，由于为食品工业提供原材料并具有很高的出口潜力，它在国家经济和社会生活中占有重要地位。多酚氧化酶是导致每年严重经济损失的酶促褐变反应的主角，从红玫瑰葡萄（Vitis vinifera L.）中对其进行了纯化和特性分析。由于其长的储存寿命和抗氧化能力，这种古老的葡萄品种具有工业加工和出口潜力。因此，从红玫瑰葡萄中纯化和生化特性分析多酚氧化酶具有重要意义。
