Polyphenol oxidases (PPOs) are important enzymes that are widely found in both prokaryotes and eukaryotes including grapes. Studies of grape PPO to date have mostly relied on enzymes extracted and purified from plants. In this work, we describe the production of the mature form of Shine Muscat grape PPO by using an Escherichia coli expression system. We have optimised the purification procedure to obtain pure and active recombinant enzymes and characterised the catalytic efficiency of the recombinant grape PPO by using ultraviolet/visible (UV/Vis) spectrophotometry. Our work provides a simple protocol of obtaining pure and active recombinant grape PPO that will enable further studies about the catalytic mechanism and inhibition of this enzyme.

中文翻译：

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一种重组生产成熟的葡萄多酚氧化酶的方法。

多酚氧化酶（PPOs）是在原核生物和真核生物（包括葡萄）中广泛发现的重要酶。迄今为止，对葡萄PPO的研究主要依赖于从植物中提取和纯化的酶。在这项工作中，我们描述了通过使用大肠杆菌表达系统生产Shine Muscat葡萄PPO的成熟形式。我们优化了纯化程序，以获得纯净和活性的重组酶，并通过紫外/可见（UV / Vis）分光光度法表征了重组葡萄PPO的催化效率。我们的工作为获得纯净和活性的重组葡萄PPO提供了一个简单的方案，这将有助于进一步研究该酶的催化机理和抑制作用。