Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/68916
標題: The role of a conserved histidine residue, His324, in Trigonopsis variabilis D-amino acid oxidase
作者: Lin, L.L.
Wang, W.C.
Ju, S.S.
Chien, H.R.
Hsu, W.H.
關鍵字: Trigonopsis variabilis
D-amino acid oxidase
FAD binding
site-specific mutagenesis
yeast rhodosporidium toruloides
catalytic
function
molecular-cloning
cephalosporin-c
kidney
expression
aspartate
sequence
binding
期刊/報告no:: Fems Microbiology Letters, Volume 176, Issue 2, Page(s) 443-448.
摘要: To investigate the functional role of an invariant histidine residue in Trigonopsis variabilis D-amino acid oxidase (DAAO), a set of mutant enzymes with replacement of the histidine residue at position 324 was constructed and their enzymatic properties were examined, Wild-type and mutant enzymes have been purified to homogeneity using the His-bound column and the molecular masses were determined to be 39.2 kDa. Western blot analysis revealed that the in vivo synthesized mutant enzymes are immune-identical with that of the wild-type DAAO. The His324Asn and His324Gln mutants displayed comparable enzymatic activity to that of the wild-type enzyme, while the other mutant DAAOs showed markedly decreased or no detectable activity. The mutants, His324/Asn/Gln/Ala/Tyr/Glu, exhibited 38-181% increase in K-m and a 2-10-fold reduction in k(cat)/K-m. Based on the crystal structure of a homologous protein, pig kidney DAAO, it is suggested that His324 might play a structural role for proper catalytic function of T. variabilis DAAO. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
URI: http://hdl.handle.net/11455/68916
ISSN: 0378-1097
文章連結: http://dx.doi.org/10.1111/j.1574-6968.1999.tb13695.x
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