8HDD

Complex structure of catalytic, small, and a partial electron transfer subunits from Burkholderia cepacia FAD glucose dehydrogenase

8HDD の概要

• エントリーDOI: 10.2210/pdb8hdd/pdb
• 関連するPDBエントリー: 6A2U
• 分子名称: Glucose dehydrogenase, Glucose dehydrogenase beta subunit, Twin-arginine translocation pathway signal, ... (6 entities in total)
• 機能のキーワード: glucose dehydrogenase, fad, burkholderia cepacia, cytochrome c, hem, oxidoreductase
• 由来する生物種: Burkholderia cepacia; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 126455.52

構造登録者

Yoshida, H.,Sode, K. (登録日: 2022-11-04, 公開日: 2022-12-14, 最終更新日: 2024-11-06)

主引用文献

Okuda-Shimazaki, J.,Yoshida, H.,Lee, I.,Kojima, K.,Suzuki, N.,Tsugawa, W.,Yamada, M.,Inaka, K.,Tanaka, H.,Sode, K.
Microgravity environment grown crystal structure information based engineering of direct electron transfer type glucose dehydrogenase.
Commun Biol, 5:1334-1334, 2022

PubMed Abstract: The heterotrimeric flavin adenine dinucleotide dependent glucose dehydrogenase is a promising enzyme for direct electron transfer (DET) principle-based glucose sensors within continuous glucose monitoring systems. We elucidate the structure of the subunit interface of this enzyme by preparing heterotrimer complex protein crystals grown under a space microgravity environment. Based on the proposed structure, we introduce inter-subunit disulfide bonds between the small and electron transfer subunits (5 pairs), as well as the catalytic and the electron transfer subunits (9 pairs). Without compromising the enzyme's catalytic efficiency, a mutant enzyme harboring Pro205Cys in the catalytic subunit, Asp383Cys and Tyr349Cys in the electron transfer subunit, and Lys155Cys in the small subunit, is determined to be the most stable of the variants. The developed engineered enzyme demonstrate a higher catalytic activity and DET ability than the wild type. This mutant retains its full activity below 70 °C as well as after incubation at 75 °C for 15 min - much higher temperatures than the current gold standard enzyme, glucose oxidase, is capable of withstanding.

PubMed: 36473944
DOI: 10.1038/s42003-022-04286-9

実験手法

X-RAY DIFFRACTION (3 Å)