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 of R-mandelate to S-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.
Bearne
S L
SL
Department of Biochemistry, Dalhousie University, Halifax, Nova Scotia, B3H 4H7, Canada.
St Maurice
M
M
Vaughan
M D
MD
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007700:Kinetics; D008333:Mandelic Acids / Q000737:chemistry / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology; D019998:Racemases and Epimerases / Q000032:analysis* / Q000378:metabolism; D015203:Reproducibility of Results; D013237:Stereoisomerism; D013379:Substrate Specificity
D008333:Mandelic Acids; D019998:Racemases and Epimerases; C021540:mandelate racemase; C037938:mandelic acid