Chitooligosaccharides are oligosaccharides with many biological activities that can be used in food production for sweeteners, preservatives and humectants, among other products. Chitin, a long-chain polymer of N-acetylglucosamine and a derivative of glucose, can be hydrolyzed by applying chitinase to break down glycosidic bonds to form chitooligosaccharides. Chitinases arising from heterologous gene expression are usually linked to a 6 × His-tag to facilitate easy purification. Heterologously expressed chitinase linked to a 6 × His-tag is a transgenic element, but enzyme activity tests cannot be used to distinguish transgenic elements from natural elements. In this study, we established a rapid and easy method to detect His-tag-containing chitinase using gold nanoparticles (AuNPs) and ssDNA aptamers. Using this method, His-tag-containing chitinase could be detected at concentrations as low as 0.136 nM within 5 min. Color changes of AuNPs showed a positive correlation with His-tag-containing chitinase concentrations.

译文

:壳寡糖是具有许多生物活性的寡糖,可用于食品生产中的甜味剂,防腐剂和保湿剂等产品。几丁质是一种N-乙酰氨基葡萄糖的长链聚合物,是葡萄糖的衍生物,可通过应用几丁质酶分解糖苷键形成壳寡糖来水解。异源基因表达产生的几丁质酶通常与6×His标签相连,以方便纯化。与6×His-tag连接的异源几丁质酶是一种转基因元件,但是酶活性测试不能用于区分转基因元件和天然元件。在这项研究中,我们建立了一种快速,简便的方法,使用金纳米颗粒(AuNPs)和ssDNA适体来检测含His标签的几丁质酶。使用这种方法,可以在5分钟内以低至0.136nM的浓度检测到含His-tag的几丁质酶。 AuNPs的颜色变化与含His标签的几丁质酶浓度呈正相关。

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