Use of a chemiluminescent detector for quantitation of nitric oxide produced in assays of denitrifying enzymes. [Achromobacter cycloblastes; Pseudomonas perfectomarini]

Conference · Fri May 01 00:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6015343

Pai, G; Payne, W J; Le Gall, J

The authors developed a closed-flow system that continuously sweeps away gases evolved in enzyme assay mixtures into a commercially available oxides of nitrogen analyzer for the quantitation of any nitric oxide (NO) present in these gases. The system is particularly useful for the study of both the stoichiometry and kinetics of NO production from nitrite by cell-free extracts of denitrifying bacteria and by the purified denitrifying nitrite reductases. In addition to its much greater sensitivity when compared to standard gas chromatographic techniques, the method offers some unique advantages since it allows measurements of initial reaction velocities, without the problems of product inhibition, which often limit kinetic studies of denitrifying enzymes. The system also allows the immediate quantitation of any NO produced and so would be useful in detecting the transient presence of this very reactive free radical, if it is produced as a free intermediate in whole cell suspensions of denitrifying bacteria respiring nitrate. The apparatus has been calibrated using a purified copper nitrite reductase from Achromobacter cycloclastes and cell-free extracts from Pseudomonas Perfectomarini, and its utility in studies of the kinetics and stoichiometry of NO production from nitrite confirmed by comparison with results obtained using manometric and gas chromatographic techniques.

• Research Organization: Univ. of Georgia, Athens
• OSTI ID: 6015343
• Report Number(s): CONF-870644-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
NITRIC OXIDE
QUANTITATIVE CHEMICAL ANALYSIS
BACTERIA
BIOCHEMICAL REACTION KINETICS
CHEMILUMINESCENCE
DENITRIFICATION
MEASURING INSTRUMENTS
NITRITES
OXIDOREDUCTASES
RADICALS
SENSITIVITY
CHALCOGENIDES
CHEMICAL ANALYSIS
CHEMICAL REACTIONS
ENZYMES
KINETICS
LUMINESCENCE
MICROORGANISMS
NITROGEN COMPOUNDS
NITROGEN OXIDES
OXIDES
OXYGEN COMPOUNDS
REACTION KINETICS
400102* - Chemical & Spectral Procedures