5zbc

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

spinqualitylabelsall models 5zbc, resolution 2.20Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5zbc is a 2 chain structure with sequence from Chromobacterium violaceum ATCC 12472. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 2.2Å Ligands: FAD, MSE Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function VIOA_CHRVO Publication Abstract from PubMed 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 A resolution. Protein engineering for improving the thermostability of tryptophan oxidase and insights from structural analysis.,Yamaguchi H, Tatsumi M, Takahashi K, Tagami U, Sugiki M, Kashiwagi T, Kameya M, Okazaki S, Mizukoshi T, Asano Y J Biochem. 2018 Nov 1;164(5):359-367. doi: 10.1093/jb/mvy065. PMID:30053101 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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