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Use of A Bioreactor Consisting of Sequentially Aligned L‐Glutamate Dehydrogenase and L‐Glutamate Oxidase for the Determination of Ammonia by Chemiluminescence
Author(s) -
Murachi T.,
Tabata M.
Publication year - 1987
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1987.tb00479.x
Subject(s) - chemiluminescence , glutamate dehydrogenase , chemistry , hydrogen peroxide , chromatography , ammonia , immobilized enzyme , luminol , ferricyanide , bioreactor , potassium ferricyanide , flow injection analysis , biochemistry , glutamate receptor , detection limit , inorganic chemistry , organic chemistry , enzyme , receptor
A chemiluminometric method for the automated flow injection analysis of ammonia is described. The essence of the invention is the use of a bioreactor consisting of both immobilized L‐glutamate dehydrogenase (GLDH) and L‐glutamate oxidase (GLXD), which are sequentially aligned in this order in a minicolumn measuring 2.0 × 20 mm. The unidirectional constant flow of liquid through the column reactor minimizes the reversed diffusion of the solutes so that the following sequence of reactions is ensured. Thus, ammonia to be determined is first transformed by GLDH into L‐glutamate, which then produces hydrogen peroxide by GLXD. Hydrogen peroxide in the effluent from the column is then determined by its chemiluminescence upon admixing with luminol and potassium ferricyanide. The present method gives linearity of the standard curve for ammonia up to 1.0 mM. It is at least 100 times more sensitive than the conventional method for ammonia assay using ultraviolet absorption measurement.