Document Detail


Coupled optical rate determinations of amino acid oxidase activity.
MedLine Citation:
PMID:  235296     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Methods are described in which liberation of ammonia from amino acid substrates by the D- and L-amino acid oxidases may be coupled with the NADH-dependent reductive amination of 2-oxoglutarate catalysed by exogenous glutamate dehydrogenase (L-glutamate: NAD oxidoreductase (deaminating), EC 1.4.1.2). The inhibition of D-amino acid oxidase (D-amino acid:O2 oxidoreductase (deaminating), EC 1.4.3.3) by ADP needed to activate and stabilise glutamate dehydrogenase was relieved by FAD, and the substrate was D-alanine at approximately 6-fold Km concentration. Neither FAD or FMN were required in the L-amino acid oxidase (L-amino acid:O2 oxidoreductase (deaminating), EC 1.4.3.2) assay; this utilised L-leucine as substrate in a concentration approximately 7-fold the Km value. The methods were reasonably sensitive and precise, and a linear relationship between activity and enzyme concentration prevailed up to an absorbance change of 0.050 per min. They have the advantage of being amenable to automation and to employment of fluorescence techniques should greater sensitivity be required.
Authors:
D J Holme; D M Goldberg
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  377     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  1975 Jan 
Date Detail:
Created Date:  1975-06-27     Completed Date:  1975-06-27     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  61-70     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Alanine
Amino Acid Oxidoreductases / analysis*
Animals
D-Amino-Acid Oxidase / analysis
Glutamate Dehydrogenase
Kidney / enzymology
Kinetics
Leucine
Manometry
Methods
NAD
NADP
Spectrophotometry
Swine
Chemical
Reg. No./Substance:
53-59-8/NADP; 53-84-9/NAD; 56-41-7/Alanine; 61-90-5/Leucine; EC 1.4.-/Amino Acid Oxidoreductases; EC 1.4.1.2/Glutamate Dehydrogenase; EC 1.4.3.3/D-Amino-Acid Oxidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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