界面活性剤フリーの高活性電極触媒を開発~構造生物電気化学に基づく酵素の合理的設計~

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2025-07-28京都大学

京都大学などの研究グループは、Gluconobacter japonicus由来のフルクトース脱水酵素(FDH)の膜結合領域を除去した変異体を開発。従来は界面活性剤を用いて精製していたが、この変異体は界面活性剤なしで精製可能で、構造変化もほぼなく酵素活性を維持。さらに、電極への高密度吸着が可能となり、直接電子移動(DET)型反応速度が野生型の14倍に向上。構造生物電気化学に基づく酵素設計の有効性を示し、バイオ燃料電池やセンサーへの応用が期待される。

界面活性剤フリーの高活性電極触媒を開発~構造生物電気化学に基づく酵素の合理的設計~
細胞膜から遊離した高活性FDH変異体によるDET型反応のイメージ(©京都大学生体機能化学研究室)

<関連情報>

界面活性剤不含のヘテロトリマー型フルクトースデヒドロゲナーゼ変異体を用いたヘム1cおよびC末端親水性領域を欠失させた改良型直接バイオ電気化学的フルクトース酸化 Improved Direct Bioelectrochemical Fructose Oxidation with Surfactant-Free Heterotrimeric Fructose Dehydrogenase Variant Truncating Heme 1c and C-Terminal Hydrophobic Regions

Taiki Adachi,Konatsu Ichikawa,Tomoko Miyata,Fumiaki Makino,Hideaki Tanaka,Keiichi Namba,and Keisei Sowa
ACS Electrochemistry  Published: July 11, 2025
DOI:https://doi.org/10.1021/acselectrochem.5c00106

Abstract

Direct electron transfer (DET)-type bioelectrocatalysis is a coupled redox reaction between enzymatic and electrode reactions. Such mediatorless reactions are an environmentally safe approach that can be applied to various bioelectrochemical devices. We focused on fructose dehydrogenase (FDH), a membrane-bound heterotrimeric enzyme that catalyzes DET-type d-fructose oxidation. Although the overall structure was recently elucidated, its membrane-bound region has not been completely identified. Therefore, this study assumed that the heme 1c region and C-terminal hydrophobic region (CHR) were bound to the membrane. A constructed double variant (Δ1cΔCHR_FDH) was soluble without any surfactants; additionally, cryo-electron microscopy confirmed that this variant was downsized. Δ1cΔCHR_FDH exhibited a 14-fold higher catalytic current density (11 ± 1 mA cm–2) than that of the wild-type recombinant FDH (rFDH) at multi-walled carbon nanotube electrodes. Kinetic analysis of the voltammograms suggested that downsizing of the enzyme and the removal of surfactants increased the surface concentration of enzymes at the electrode. This study will lead to efficient bioelectrocatalysis, overcoming the interference of surfactants.

生物化学工学
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