Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/68903
標題: A conserved aspartate is essential for FAD binding and catalysis in the D-amino acid oxidase from Trigonopsis variabilis
作者: Ju, S.S.
Lin, L.L.
Wang, W.C.
Hsu, W.H.
關鍵字: D-amino acid oxidase
site-directed mutagenesis
flavin adenine
dinucleotide binding
Trigonopsis variabilis
flavin adenine-dinucleotide
site-specific mutagenesis
cephalosporin-c
rhodotorula-gracilis
phenotypic selection
phenol hydroxylase
molecular-cloning
sequence-analysis
kidney
dehydrogenase
期刊/報告no:: Febs Letters, Volume 436, Issue 1, Page(s) 119-122.
摘要: To evaluate the possible contribution of Asp(206) of Trigonopsis variabilis D-amino acid oxidase (DAO) to its flavin adenine dinucleotide (FAD) binding and catalytic function, six mutant enzymes were constructed by site-directed mutagenesis. Western immunoblot analysis revealed that a protein with an apparent molecular mass of about 39.2 kDa was present in the cell-free extracts of wild-type and mutant strains. Replacement of Asp(206) with Leu, Gly, and Asn resulted in the loss of DAO activity and characteristic absorption spectrum for flavoenzyme, while the other mutant DAOs, Asp(206)Glu, Asp(206)Ser, and Asp(206)Ala, exhibited a similar spectral profile to that of wildtype enzyme and retained about 6-90% of the enzyme activity. These results suggested that Asp(206) of T. variabilis DAO might play an important role in the binding of FAD. (C) 1998 Federation of European Biochemical Societies.
URI: http://hdl.handle.net/11455/68903
ISSN: 0014-5793
文章連結: http://dx.doi.org/10.1016/s0014-5793(98)01108-9
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