Freezing-directed construction of enzyme/nano interfaces: Reagentless conjugation, superior activity and better stability
Freezing-directed construction of enzyme/nano interfaces: Reagentless conjugation, superior activity and better stability
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DOI：
10.1016/j.cclet.2023.108894
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发表时间：
2023-08
影响因子：
9.1
通讯作者：
Ke Quan;Jiajie Tong;Lifang Chen;Shuyao Fang;Meng-shi Li;Linlin Wu;Zhihe Qing
中科院分区：
文献类型：
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作者：
Ke Quan;Jiajie Tong;Lifang Chen;Shuyao Fang;Meng-shi Li;Linlin Wu;Zhihe Qing
将酶固定在纳米界面上是促进酶工业化应用的一个有利策略。尽管由于金纳米颗粒（AuNPs）迷人的物理化学性质，人们一直致力于使用其作为缀合基质，但在保持酶活性的同时避免繁琐的修饰仍然是一个艰巨的挑战。本文中，酶/纳米界面的冷冻定向缀合在没有额外试剂的情况下构建。作为概念的证明，选择葡萄糖氧化酶（GOx）作为模型酶。一锅法结合过程可以在− 20 ° C下在水溶液中轻松完成。此外，与GOx的负载在金纳米粒子在冷冻，酶表现出优越的催化活性和稳定性后，热和pH值的扰动。然后详细讨论了活性增强的机制。结果表明，冷冻条件下较高的装载密度和较多的酶倾向于通过Au-S键结合AuNPs是其上级活性的主要因素。更重要的是，该方法具有普适性，也可应用于其他含有天然半胱氨酸的酶，如辣根过氧化物酶（HRP）和木瓜蛋白酶。这种简单的结合策略伴随着显着的生物活性，扩大了酶生物传感，微器件，甚至药物输送的可能性。
Immobilizing enzyme to nano interfaces has demonstrated to be a favorable strategy for prompting the industrialized application of enzyme. Despite tremendous endeavor has been devoted to using gold nanoparticles (AuNPs) as conjugation matrix due to its fascinating physico-chemical properties, maintaining enzymatic activity while circumventing cumbersome modification remains a formidable challenge. Herein, the freezing-directed conjugation of enzyme/nano interfaces was constructed without extra reagent. As the proof of concept, glucose oxidase (GOx) was chosen as model enzyme. The one-pot conjugation process can be facilely completed at −20 °C under aqueous solution. Moreover, with the loading of GOx on AuNP at freezing, the enzyme exhibited superior catalytic activity and stability upon thermal and pH perturbation. The mechanism of boosted activity was then discussed in detail. It was found that higher loading density under freezing condition and more enzyme tending to bind AuNPsviaAu-S bond were the main factors for the superior activity. More importantly, this methodology was universal and can also be applied to other enzyme which contains natural cysteine, such as horseradish peroxidase (HRP) and papain. This facile conjugation strategy accompanied by remarkable bioactivity expand the possibilities for enzymatic biosensing, microdevice and even drug delivery.