Engineered fungus derived FAD-dependent glucose dehydrogenase with acquired ability to utilize hexaammineruthenium(III) as an electron acceptor
| Autor: | Koji Sode, Nanami Suzuki, Katsuhiro Kojima, Noya Loew, Madoka Okurita, Wakako Tsugawa, David C. Klonoff, Kazushige Mori, Hiromi Yoshida |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Models Molecular Biophysics Enzyme electrode Electrons Electrochemistry Protein Engineering 01 natural sciences Redox 03 medical and health sciences chemistry.chemical_compound Glucose dehydrogenase Glucose oxidase Physical and Theoretical Chemistry chemistry.chemical_classification biology Chemistry 010401 analytical chemistry Glucose 1-Dehydrogenase General Medicine Maltose Electrochemical Techniques Electron acceptor Combinatorial chemistry 0104 chemical sciences 030104 developmental biology Amino Acid Substitution biology.protein Flavin-Adenine Dinucleotide Ruthenium Compounds Ferricyanide Aspergillus flavus |
| Zdroj: | Bioelectrochemistry (Amsterdam, Netherlands). 123 |
| ISSN: | 1878-562X |
| Popis: | Fungal FAD-dependent glucose dehydrogenases (FADGDHs) are considered to be superior enzymes for glucose sensor strips because of their insensitivity to oxygen and maltose. One highly desirable mediator for enzyme sensor strips is hexaammineruthenium(III) chloride because of its low redox potential and high storage stability. However, in contrast to glucose oxidase (GOx), fungal FADGDH cannot utilize hexaammineruthenium(III) as electron acceptor. Based on strategic structure comparison between FADGDH and GOx, we constructed a mutant of Aspergillus flavus-derived FADGDH, capable of utilizing hexaammineruthenium(III) as electron acceptor: AfGDH-H403D. In AfGDH-H403D, a negative charge introduced at the pathway-entrance leading to the FAD attracts the positively charged hexaammineruthenium(III) and guides it into the pathway. The corresponding amino acid in wild-type GOx is negatively charged, which explains the ability of GOx to utilize hexaammineruthenium(III) as electron acceptor. Electrochemical measurements showed a response current of 46.0 μA for 10 mM glucose with AfGDH-H403D and hexaammineruthenium(III), similar to that with wild-type AfGDH and ferricyanide (47.8 μA). Therefore, AfGDH-H403D is suitable for constructing enzyme electrode strips with hexaammineruthenium(III) chloride as sole mediator. Utilization of this new, improved fungal FADGDH should lead to the development of sensor strips for blood glucose monitoring with increased accuracy and less stringent packing requirements. |
| Databáze: | OpenAIRE |
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