Enzymatic cycling assay for phenylpyruvate☆

Add time:08/26/2019    Source:sciencedirect.com

Enzymatic cycling assays for the determination of l-phenylalanine and phenylpyruvate in deproteinized tissue extracts are described. Assay 1 couples glutamine transaminase K with l-phenylalanine dehydrogenase. Assay 2 combines phenylalanine dehydrogenase, l-amino acid oxidase, and catalase. In both assays, tyrosine and some other amino acids (or their α-keto acid analogs) can replace phenylalanine (or phenylpyruvate) to a small extent. Thus, if phenylalanine is to be measured a correction must be made for the nonspecificity of the reaction. By removing phenylalanine on a cation-exchange column it was possible to measure phenylpyruvate in tissue extracts. Concentrations of phenylpyruvate (μmol/kg) in normal rat liver, kidney, and brain were 2.1 ± 1.1 (n = 8), 1.8 ± 0.4 (n = 4), and 3.3 ± 0.6 (n = 4), respectively.

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