An Assay for Mandelate Racemase Using High-Performance Liquid Chromatography

Mandelate racemase (EC 5.1.2.2) catalyzes the interconversion of the two stereoisomers of mandelic acid. A fixed-time assay for the quantification of mandelate racemase activity has been developed. The assay involves enzymatic conversion ofR-mandelate toS-mandelate (or the reverse reaction) followed...

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Veröffentlicht in:	Analytical biochemistry 1999-05, Vol.269 (2), p.332-336
Hauptverfasser:	Bearne, Stephen L., St. Maurice, Martin, Vaughan, Mark D.
Format:	Artikel
Sprache:	eng
Schlagworte:	assay Chromatography, High Pressure Liquid - methods HPLC Kinetics mandelate Mandelic Acids - chemistry Mandelic Acids - metabolism Pseudomonas putida - enzymology racemase Racemases and Epimerases - analysis Racemases and Epimerases - metabolism Reproducibility of Results Stereoisomerism Substrate Specificity
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Beschreibung
Zusammenfassung:	Mandelate racemase (EC 5.1.2.2) catalyzes the interconversion of the two stereoisomers of mandelic acid. A fixed-time assay for the quantification of mandelate racemase activity has been developed. The assay involves enzymatic conversion ofR-mandelate toS-mandelate (or the reverse reaction) followed by separation and detection of the substrate and product using isocratic reversed-phase high-performance liquid chromatography on a Sumichiral OA-6100 column and absorbance detection. This method offers an economical and efficient alternative to the existing circular dichroism-based and coupled assays.
ISSN:	0003-2697 1096-0309
DOI:	10.1006/abio.1999.4018