Interaction of Flavin-Dependent Fructose Dehydrogenase with Cytochrome c as Basis for the Construction of Biomacromolecular Architectures on Electrodes.
Interaction of Flavin-Dependent Fructose Dehydrogenase with Cytochrome c as Basis for the Construction of Biomacromolecular Architectures on Electrodes.
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DOI：
10.1021/acs.analchem.6b00815
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发表时间：
2016-06
影响因子：
7.4
通讯作者：
Chris Wettstein;K. Kano;D. Schäfer;U. Wollenberger;F. Lisdat
中科院分区：
文献类型：
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作者：
Chris Wettstein;K. Kano;D. Schäfer;U. Wollenberger;F. Lisdat
基于酶和氧化还原蛋白质在电极表面上的定义排列的电子转移（ET）链的创建代表了生物电催化领域内的一种有趣的方法。在这项研究中，我们研究了黄素依赖性酶果糖脱氢酶（FDH）与氧化还原蛋白细胞色素c（cyt c）的ET反应。在酸性和中性溶液中，发现反应的两个不同的pH最佳值。当细胞色素C被吸附在电极表面上，而酶仍然在溶液中，ET进行有效地在中性pH的介质中。蛋白间ET也观察到在酸性介质中，但是，它似乎是效率较低。这些发现表明，两种不同的酶和细胞色素c之间的ET途径可能会发生。此外，细胞色素c和FDH固定在多层的电极表面上的另一种生物大分子：DNA（双链）使用层层技术。的双蛋白多层架构显示依赖于果糖浓度的催化反应，表明两种蛋白质之间的ET反应是可行的，即使在固定化状态。该电极对果糖有较好的响应，并具有良好的储存稳定性。我们的研究结果有助于更好地理解FDH和细胞色素c之间的ET反应，并提供了基础，为创建全生物分子为基础的果糖传感器的灵敏度可以控制的层制备。
The creation of electron transfer (ET) chains based on the defined arrangement of enzymes and redox proteins on electrode surfaces represents an interesting approach within the field of bioelectrocatalysis. In this study, we investigated the ET reaction of the flavin-dependent enzyme fructose dehydrogenase (FDH) with the redox protein cytochrome c (cyt c). Two different pH optima were found for the reaction in acidic and neutral solutions. When cyt c was adsorbed on an electrode surface while the enzyme remained in solution, ET proceeded efficiently in media of neutral pH. Interprotein ET was also observed in acidic media; however, it appeared to be less efficient. These findings suggest that two different ET pathways between the enzyme and cyt c may occur. Moreover, cyt c and FDH were immobilized in multiple layers on an electrode surface by means of another biomacromolecule: DNA (double stranded) using the layer-by-layer technique. The biprotein multilayer architecture showed a catalytic response in dependence on the fructose concentration, indicating that the ET reaction between both proteins is feasible even in the immobilized state. The electrode showed a defined response to fructose and a good storage stability. Our results contribute to the better understanding of the ET reaction between FDH and cyt c and provide the basis for the creation of all-biomolecule based fructose sensors the sensitivity of which can be controlled by the layer preparation.