Constructive Optimization of a Multienzymatic Film Based on a Cascade Reaction for Electrochemical Biosensors
Constructive Optimization of a Multienzymatic Film Based on a Cascade Reaction for Electrochemical Biosensors
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DOI：
10.1021/acsomega.0c05521
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发表时间：
2020-12-08
期刊：
影响因子：
4.1
通讯作者：
Tokito S
中科院分区：
文献类型：
--
作者：
Sasaki K;Furusawa H;Nagamine K;Tokito S
多酶级联反应在电化学生物传感器中的应用除了在电极上选择性地结合多个酶之外，还具有扩展目标底物的优点。然而，多酶系统具有物质转化效率低的缺点，这是因为中间体在酶之间的传递耗时和/或中间体的扩散损失。在这项研究中，一个多酶膜的氨检测传感器的最佳结构进行了研究使用的级联反应的l-谷氨酸氧化酶和l-谷氨酸脱氢酶作为模型传感器。制备了三种酶膜：（1）混合膜，其设计为在紧密定位的酶之间具有短的扩散距离，（2）正常顺序分层膜，其布置用于正确的反应途径，以及（3）反向顺序分层膜作为阴性对照。随后使用普鲁士蓝电极的时间依赖性电位测量来比较氨向过氧化氢的转化效率，从而确定过氧化氢的量。结果表明，在三种酶膜中，正序分层膜的转化效率最高。对级联反应中间转化效率的定量评价表明，与混合膜（34%）相比，在第一酶促反应步骤中获得了92%的更高转化效率。这些发现将促进多酶级联反应系统不仅在生物传感器和生物反应器中，而且在各个工业领域的使用。
The application of a multienzyme cascade reaction in electrochemical biosensors has the advantage of expanding the target substrates in addition to selectivity combining multiple enzymes on an electrode. However, the multienzyme system has the drawback of inefficient substance conversion because of the time-consuming passing of intermediates between the enzymes and/or diffusional loss of the intermediates. In this study, the optimal construction of a multienzymatic film in an ammonia detection sensor was investigated using a cascade reaction of l-glutamate oxidase and l-glutamate dehydrogenase as a model sensor. Three enzymatic films were prepared: (1) a mixed film designed to have a short diffusional distance between closely located enzymes, (2) a normal-sequential layered film arranged for the correct reaction pathway, and (3) a reverse-sequential layered film as a negative control. This was followed by comparison of the conversion efficiency of ammonia to hydrogen peroxide using time-dependent potentiometric measurements of a Prussian blue electrode determining the hydrogen peroxide amount. The results indicate that the conversion efficiency of the normal-sequential layered film was the highest among the three enzymatic films. The quantitative evaluation of the intermediate conversion efficiency of the cascade reaction showed that compared to the mixed film (34%), a higher conversion efficiency of 92% was obtained in the first enzymatic reaction step. These findings will promote the use of multienzymatic cascade reaction systems not only in biosensors and bioreactors but also in various industrial fields.
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影响因子：
4.6
作者：
Shiwaku R;Matsui H;Nagamine K;Uematsu M;Mano T;Maruyama Y;Nomura A;Tsuchiya K;Hayasaka K;Takeda Y;Fukuda T;Kumaki D;Tokito S
通讯作者：
Tokito S
影响因子：
2.7
作者：
Arima, J;Tamura, T;Inagaki, K
通讯作者：
Inagaki, K
影响因子：
2.8
作者：
COULTON, JW;KAPOOR, M
通讯作者：
KAPOOR, M
影响因子：
4.3
作者：
Hildebrand, Falk;Luetz, Stephan
通讯作者：
Luetz, Stephan
影响因子：
16.6
作者：
Elani, Yuval;Law, Robert V.;Ces, Oscar
通讯作者：
Ces, Oscar