Improvement in Thermal Stability and Substrate Binding of Pig Kidney D-Amino Acid Oxidase by Chemical Modification

2004 ◽  
Vol 112 (3) ◽  
pp. 123-132 ◽  
Author(s):  
Mikio Bakke ◽  
Naoki Kajiyama
Keyword(s):  
Thermal Stability ◽  
Amino Acid ◽  
Chemical Modification ◽  
Substrate Binding ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Pig Kidney

scholarly journals Studies on the active centre of Rhodotorula gracilisd-amino acid oxidase and comparison with pig kidney enzyme

1992 ◽  
Vol 286 (2) ◽  
pp. 389-394 ◽  
Author(s):  
L Pollegioni ◽  
S Ghisla ◽  
M S Pilone
Keyword(s):  
Amino Acid ◽  
Solvent Accessibility ◽  
Alkylating Agents ◽  
Fine Tuning ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Active Centre ◽  
Pig Kidney ◽  
Mammalian Enzyme ◽  
Active Centres

D-Amino acid oxidase (EC 1.4.3.3) from Rhodotorula gracilis has been reconstituted with 8-chloro-, 8-mercapto-, 6-hydroxy-, 2-thio-, 5-deaza- and 1-deaza-FAD, and the properties of the resulting complexes have been studied and compared with those of the correspondingly modified pig kidney D-amino acid oxidases. Binding appears to be tight for most analogues, at least as tight as for native FAD (approximately 10(-8) M). 8-Mercapto- and 6-hydroxy-FAD bind in their para- and ortho-quinoid forms respectively to yeast D-amino acid oxidase, inferring the presence of a positive charge near the flavin N(1) position, as in the case of the mammalian enzyme. On the other hand, important differences in active-site microenvironment emerge: solvent accessibility to flavin position 8 is drastically restricted in yeast D-amino acid oxidase as indicated by the unreactivity of 8-chloro- and 8-mercapto-FAD enzyme with thiolates and alkylating agents. Significantly different microenvironments are also likely to occur around the flavin positions N(1)-C(2) = 0, N(3)-H and N(5). This is deduced from the differences in interaction of the two proteins with 1-deaza-FAD, 5-deaza-FAD and 2-thio-FAD and from the properties of the respective complexes. The same re-side flavin stereospecificity as shown by the mammalian enzyme was determined for the yeast enzyme using 8-hydroxy-5-deaza-FAD. Thus we can deduce the presence of a similar pattern of functional groups at the active centres of the two enzymes, while the fine tuning of specificity and regulation correlate with environmental differences at specific flavin loci.

scholarly journals Contribution of the dimeric state to the thermal stability of the flavoprotein D-amino acid oxidase

2003 ◽  
Vol 12 (5) ◽  
pp. 1018-1029 ◽  
Author(s):  
Loredano Pollegioni ◽  
Stefania Iametti ◽  
Dimitrios Fessas ◽  
Laura Caldinelli ◽  
Luciano Piubelli ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Amino Acid ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Thermal Stability Of
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