Electroactive Multilayer Assemblies of Bilirubin Oxidase and Human Cytochrome C Mutants: Insight in Formation and Kinetic Behavior

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• 作者：Franziska Wegerich ; Paola Turano ; Marco Allegrozzi ; Helmuth Mo虉hwald ; Fred Lisdat
• 刊名：Langmuir
• 出版年：2011
• 出版时间：April 5, 2011
• 年：2011
• 卷：27
• 期：7
• 页码：4202-4211
• 全文大小：1132K
• 年卷期：v.27,no.7(April 5, 2011)
• ISSN：1520-5827

文摘

Here, we report on cytochrome c/bilirubin oxidase multilayer electrodes with different cytochrome c (cyt c) forms including mutant forms of human cyt c, which exhibit different reaction rates with bilirubin oxidase (BOD) in solution. The multilayer formation via the layer-by-layer technique and the kinetic behavior of the mono (only cyt c) and biprotein (cyt c and BOD) multilayer systems are studied by SPR and cyclic voltammetry. For the layer construction, sulfonated polyaniline is used. The only cyt c containing multilayer electrodes show that the quantity of deposited protein and the kinetic behavior depend on the cyt c form incorporated. In the case of the biprotein multilayer with BOD, it is demonstrated that the catalytic signal chain from the electrode via cyt c to BOD and oxygen can be established with all chosen cyt c forms. However, the magnitude of the catalytic current as well as the kinetic behavior differ significantly. We conclude that the different cytochrome c forms affect three parameters, identified here, to be important for the functionality of the multilayer system: the amount of molecules per layer, which can be immobilized on the electrodes, the cyt c self-exchange rate, and the rate constant for the reaction with BOD.