5ZBC

Crystal structure of Se-Met tryptophan oxidase (C395A mutant) from Chromobacterium violaceum

5ZBC の概要

• エントリーDOI: 10.2210/pdb5zbc/pdb
• 分子名称: Flavin-dependent L-tryptophan oxidase VioA, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• 機能のキーワード: oxidase, oxidoreductase
• 由来する生物種: Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 / JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 103652.93

構造登録者

Yamaguchi, H.,Tatsumi, M.,Takahashi, K.,Tagami, U.,Sugiki, M.,Kashiwagi, T.,Okazaki, S.,Mizukoshi, T.,Asano, Y. (登録日: 2018-02-11, 公開日: 2018-12-19, 最終更新日: 2024-11-06)

主引用文献

Yamaguchi, H.,Tatsumi, M.,Takahashi, K.,Tagami, U.,Sugiki, M.,Kashiwagi, T.,Kameya, M.,Okazaki, S.,Mizukoshi, T.,Asano, Y.
Protein engineering for improving the thermostability of tryptophan oxidase and insights from structural analysis.
J. Biochem., 164:359-367, 2018

PubMed Abstract: l-Tryptophan oxidase, VioA from Chromobacterium violaceum, which has a high substrate specificity for tryptophan, is useful for quantitative assay of tryptophan. However, stability of wild type VioA is not enough for its application in clinical or industrial use. To improve the thermal stability of the enzyme, we developed a VioA (C395A) mutant, with higher stability than wild type VioA. The VioA (C395A) exhibited similar specificity and kinetic parameter for tryptophan to wild type. Conventionally, the quantity of tryptophan is determined by instrumental methods, such as high-performance liquid chromatography (HPLC) after pre-column-derivatization. Using the mutant enzyme, we succeeded in the tryptophan quantification in human plasma samples, to an accuracy of <2.9% when compared to the instrumental method, and to a precision of CV <3.2%. To analyse the improvement in storage stability and substrate specificity, we further determined the crystal structures of VioA (C395A) complexed with FAD, and with FAD and tryptophan at 1.8 Å resolution.

PubMed: 30053101
DOI: 10.1093/jb/mvy065

実験手法

X-RAY DIFFRACTION (2.2 Å)