5ZBD
Crystal structure of tryptophan oxidase (C395A mutant) from Chromobacterium violaceum
Summary for 5ZBD
| Entry DOI | 10.2210/pdb5zbd/pdb |
| Descriptor | Flavin-dependent L-tryptophan oxidase VioA, TRYPTOPHAN, FLAVIN-ADENINE DINUCLEOTIDE, ... (4 entities in total) |
| Functional Keywords | oxidase, oxidoreductase |
| Biological source | Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 / JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757) |
| Total number of polymer chains | 2 |
| Total formula weight | 102935.91 |
| Authors | Yamaguchi, H.,Tatsumi, M.,Takahashi, K.,Tagami, U.,Sugiki, M.,Kashiwagi, T.,Okazaki, S.,Mizukoshi, T.,Asano, Y. (deposition date: 2018-02-11, release date: 2018-12-19, Last modification date: 2024-03-27) |
| Primary citation | 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 Cited by 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: 30053101DOI: 10.1093/jb/mvy065 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.8 Å) |
Structure validation
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