Enzyme Assay by Repetitive Flow Injection Analysis. Application to the Assay of Hog Kidney Aminoacylase

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Abstract
The initial rate of hydrolysis of N-acetylmethionine by hog kidney aminoacylase (EC 3.5.1.14) was determined by repetitive flow injection analysis. The reaction mixture was slowly pumped through an injection valve from which portions were injected into a reagent stream consisting of o-phthaldialdehyde and mercaptoethanol in a borate buffer. This reagent converted the product methionine into a spectrophotometrically-detected isoindole. In a typical assay 15 data points were obtained within the first 22 min of reaction. Initial rates obtained by the method were equal to those obtained by ninhydrin assay.