7E0C

Structure of L-glutamate oxidase R305E mutant

7E0C の概要

• エントリーDOI: 10.2210/pdb7e0c/pdb
• 関連するPDBエントリー: 2e1m
• 分子名称: L-glutamate oxidase, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• 機能のキーワード: l-glutamic acid oxidase, l-amino acid oxidase, oxidoreductase
• 由来する生物種: Streptomyces sp. X-119-6
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 77212.80

構造登録者

Ito, N.,Matsuo, S.,Inagaki, K.,Imada, K. (登録日: 2021-01-27, 公開日: 2021-04-07, 最終更新日: 2023-11-29)

主引用文献

Yano, Y.,Matsuo, S.,Ito, N.,Tamura, T.,Kusakabe, H.,Inagaki, K.,Imada, K.
A new l-arginine oxidase engineered from l-glutamate oxidase.
Protein Sci., 30:1044-1055, 2021

PubMed Abstract: The alternation of substrate specificity expands the application range of enzymes in industrial, medical, and pharmaceutical fields. l-Glutamate oxidase (LGOX) from Streptomyces sp. X-119-6 catalyzes the oxidative deamination of l-glutamate to produce 2-ketoglutarate with ammonia and hydrogen peroxide. LGOX shows strict substrate specificity for l-glutamate. Previous studies on LGOX revealed that Arg305 in its active site recognizes the side chain of l-glutamate, and replacement of Arg305 by other amino acids drastically changes the substrate specificity of LGOX. Here we demonstrate that the R305E mutant variant of LGOX exhibits strict specificity for l-arginine. The oxidative deamination activity of LGOX to l-arginine is higher than that of l-arginine oxidase form from Pseudomonas sp. TPU 7192. X-ray crystal structure analysis revealed that the guanidino group of l-arginine is recognized not only by Glu305 but also Asp433, Trp564, and Glu617, which interact with Arg305 in wild-type LGOX. Multiple interactions by these residues provide strict specificity and high activity of LGOX R305E toward l-arginine. LGOX R305E is a thermostable and pH stable enzyme. The amount of hydrogen peroxide, which is a byproduct of oxidative deamination of l-arginine by LGOX R305E, is proportional to the concentration of l-arginine in a range from 0 to 100 μM. The linear relationship is maintained around 1 μM of l-arginine. Thus, LGOX R305E is suitable for the determination of l-arginine.

PubMed: 33764624
DOI: 10.1002/pro.4070

実験手法

X-RAY DIFFRACTION (2.65 Å)