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dc.creatorIgnjatović, Svetlana
dc.creatorMajkić-Singh, Nada
dc.date.accessioned2019-09-02T10:48:50Z
dc.date.available2019-09-02T10:48:50Z
dc.date.issued1994
dc.identifier.issn0003-2670
dc.identifier.urihttp://farfar.pharmacy.bg.ac.rs/handle/123456789/115
dc.description.abstractA simple kinetic method for assay of d-amino acid oxidase (d-AAO) catalytic activity using 2,2′-azinodi(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) as chromogen is described. Hydrogen peroxide produced in the reaction catalysed by d-AAO stoichiometrically oxidezes ABTS in a peroxidase-catalysed reaction. The absorbance of the chromogen measured at 410 nm is directly proportional to d-AAO activity. The estimated values of the Michaelis-Menten constants (Km) for d-alanine, d-proline, d-valine, d-2-aminobutyrate and d-leucine, determined in 100 mM pyrophosphate buffer (pH 8.8) at 30°C, were 2.60, 3.40, 4.60, 1.98 and 0.14 mM, respectively. The pH optima for the d-AAO activity was found to be between 8.6 and 8.8. The optimum temperature was about 30°C and the activation energy 8.89 kJ mol-1. The Km values for peroxidase, as an auxiliary enzyme in the indicator reaction, were 19.0, 20.8, 17.3 and 27.2 U 1-1 for d-alanine, d-proline, d-2-aminobutyrate and d-leucine, respectively, as substrate. The Km value for ABTS was 1.16 mM. The inhibition by sodium benzoate was found to be competitive with an inhibition constant, Ki, of 2.7 × 10-5 M. In all experiments, 50.5 μM of free flavine adenine dinucleptide was added to the reaction mixture to avoid dissociation of the holoenzyme.en
dc.rightsrestrictedAccess
dc.sourceAnalytica Chimica Acta
dc.subjectAmino acid oxidaseen
dc.subjectCatalytic activityen
dc.subjectEnzymatic methodsen
dc.subjectKinetic methodsen
dc.titleStudy of the kinetic characteristics of d-amino acid oxidase using 2,2′-azinodi(3-ethylbenzothiazoline-6-sulphonic acid) as oxygen acceptoren
dc.typearticle
dc.rights.licenseARR
dcterms.abstractИгњатовић, Светлана; Мајкић-Сингх, Нада;
dc.citation.volume285
dc.citation.issue3
dc.citation.spage369
dc.citation.epage375
dc.citation.other285(3): 369-375
dc.identifier.doi10.1016/0003-2670(94)80078-2
dc.identifier.scopus2-s2.0-0028010749
dc.identifier.rcubconv_5471
dc.type.versionpublishedVersion


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