Characterization and application of novel dye-linked L-glutamate dehydrogenase

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K05834
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 38030:Applied biochemistry-related
• Research Institution: University of Fukui
• Principal Investigator: Satomura Takenori 福井大学, 学術研究院工学系部門, 教授 (50412317)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 酸化還元酵素 / L-グルタミン酸 / バイオセンサ

Outline of Final Research Achievements

The dye-linked L-glutamate dehydrogenase (Dye-GluDH) is an enzyme that catalyzes the oxidation of L-glutamate in the presence of artificial electron acceptors. This enzyme can transfer electrons from the substrate to an electrode via an artificial electron acceptor, making it possible to use as an element for an L-glutamate biosensor. However, the existence of an enzyme showing Dye-GluDH activity has not been reported so far. Therefore, we have screened Dye-GluDH in various microorganisms. As a result, we successfully detected Dye-GluDH activity in the thermophilic bacterium Geobacillus kaustphilus. We also succeeded in the construction of a recombinant protein expression system for G. kaustphilus Dye-GluDH in Escherichia coli, analysis of the detailed enzymatic properties, and evaluation of the Dye-GuDH as an element for L-glutamate biosensor.

Free Research Field

応用生物化学

Academic Significance and Societal Importance of the Research Achievements

本申請で見出したDye-GluDHは、L-グルタミン酸を電気化学的に測定可能なバイオセンサを開発するうえで非常に高い有用性を示した。L-グルタミン酸は、食品の品質管理に有用なインジケーター、また、様々な病気に対する重要なバイオマーカーになることが知られている。これまでに、L-グルタミン酸を測定可能な最適なバイオセンサ用素子は、見出されていなかった。本申請で見出したDye-GluDHを用いてバイオセンサを構築できれば、これまでには無い簡便で高感度にL-グルタミン酸測定用のバイオデバイスの開発が可能となる。