The rapid and specific determination of picomole quantities of 8-aminolevulinate has been accomplished by its enzymatic conversion to uroporphyrinogen I and subsequent fluorometric detection of the oxidized uroporphyrin I. The coupled-enzyme assay was linear with time and protein concentration and required less than 3 h for 25 individual determinations. Under the standard assay conditions, 5-100 pmol of uroporphyrin I was reliably quantitated; these values corresponded to a range of δ-aminolevulinate synthase activities from 0.2 to 15 nmol/h/ml enzyme. The sensitivity of this method was comparable to the more time-consuming radiochemical determinations of δ-aminolevulinate synthase activity. Theδ-aminolevulinate synthase activity of liver homogenates from uninduced rats was 8.7 U/g liver (37 °C).

Keywords:
δ-Aminolevulinate synthase, Fluorometric assay, δ-Aminolevulinate dehydratase, Porphobilinogen deaminase
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1982
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