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首页> 外文期刊>Chemistry: A European journal >Real-time monitoring of mass-transport-related enzymatic reaction kinetics in a nanochannel-array reactor
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Real-time monitoring of mass-transport-related enzymatic reaction kinetics in a nanochannel-array reactor

机译：实时监测纳米通道阵列反应器中与运输有关的酶反应动力学

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摘要

To understand the fundamentals of enzymatic reactions confined in micro-anosystems, the construction of a small enzyme reactor coupled with an integrated real-time detection system for monitoring the kinetic information is a significant challenge. Nano-enzyme array reactors were fabricated by covalently linking enzymes to the inner channels of a porous anodic alumina (PAA) membrane. The mechanical stability of this nanodevice enables us to integrate an electrochemical detector for the real-time monitoring of the formation of the enzyme reaction product by sputtering a thin Pt film on one side of the PAA membrane. Because the enzymatic reaction is confined in a limited nanospace, the mass transport of the substrate would influence the reaction kinetics considerably. Therefore, the oxidation of glucose by dissolved oxygen catalyzed by immobilized glucose oxidase was used as a model to investigate the mass-transport-related enzymatic reaction kinetics in confined nanospaces. The activity and stability of the enzyme immobilized in the nanochannels was enhanced. In this nano-enzyme reactor, the enzymatic reaction was controlled by mass transport if the flux was low. With an increase in the flux (e.g., >50 μLmin~(-1)), the enzymatic reaction kinetics became the rate-determining step. This change resulted in the decrease in the conversion efficiency of the nano-enzyme reactor and the apparent Michaelis-Menten constant with an increase in substrate flux. This nanodevice integrated with an electrochemical detector could help to understand the fundamentals of enzymatic reactions confined in nanospaces and provide a platform for the design of highly efficient enzyme reactors. In addition, we believe that such nanodevices will find widespread applications in biosensing, drug screening, and biochemical synthesis.
机译：为了了解限制在微/纳米系统中的酶促反应的基本原理，构建一个小型酶反应器以及一个集成的实时检测系统以监测动力学信息是一个重大挑战。纳米酶阵列反应器是通过将酶与多孔阳极氧化铝（PAA）膜的内部通道共价连接而制得的。这种纳米器件的机械稳定性使我们能够集成电化学检测器，以通过在PAA膜的一侧溅射一层Pt薄膜来实时监测酶反应产物的形成。因为酶促反应被限制在有限的纳米空间中，所以底物的质量传递将显着影响反应动力学。因此，以固定化葡萄糖氧化酶催化的溶解氧对葡萄糖的氧化反应为模型，研究了在受限的纳米空间中与运输相关的酶促反应动力学。固定在纳米通道中的酶的活性和稳定性得到增强。在该纳米酶反应器中，如果通量低，则通过质量传递来控制酶反应。随着通量的增加（例如，>50μLmin〜（-1）），酶​​促反应动力学成为决定速率的步骤。这种变化导致纳米酶反应器的转化效率降低，并且表观米氏（Michaelis-Menten）常数随着底物通量的增加而降低。这种与电化学检测器集成在一起的纳米器件可以帮助理解限制在纳米空间中的酶促反应的基础，并为设计高效酶反应器提供平台。此外，我们相信，此类纳米器件将在生物传感，药物筛选和生化合成中找到广泛的应用。

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