Bi-Enzyme Alcohol Biosensors Based on Genetically Engineered Alcohol Oxidase and Different Peroxidases

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• 作者：Inna S. Alpeeva ; Aiste Vilkanauskyte ; Bertrand Ngounou ; Elisabeth Cs?regi ; Ivan Yu. Sakharov ; Mykhailo Gonchar and Wolfgang Schuhmann
• 关键词：Key words ; Bi ; enzyme biosensor ; peroxidase ; alcohol oxidase ; redox polymers ; electrodeposition paint.
• 刊名：Microchimica Acta
• 出版年：2005
• 出版时间：November 2005
• 年：2005
• 卷：152
• 期：1-2
• 页码：21-27
• 全文大小：228 KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Inorganic Chemistry
  Physical Chemistry
  Characterization and Evaluation Materials
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Wien
• ISSN：1436-5073

文摘

We report on the development of a bi-layer bi-enzyme biosensor architecture using different peroxidases and alcohol oxidase from Hansenula polymorpha C-105 as biological recognition elements. The sensor architecture comprises a first layer containing either horseradish peroxidase or royal palm tree peroxidase crosslinked with an Osmium complex-modified redox hydrogel. On top, a second layer was formed by electrochemically induced precipitation of a cathodic electrodeposition paint simultaneously entrapping alcohol oxidase isolated from a genetically modified strain of Hansenula polymorpha C-105. The sensor architecture was optimized with respect to effective electron transfer and stability of the enzyme. The main characteristics of the biosensors are an apparent maximal current I_maxapp of 572–940?nA, an apparent Michaelis constant K_Mapp of 9.5?mM, a sensitivity of 60–98?nA?mM?1 and an improved operational stability represented by a deactivation constant of 1.5–2.0 × 10?4min?1.