Abstract
The simple and sensitive spectrophotometric method for determination of branched- chain L-amino acids and their keto analogs with leucine dehydrogenase (EC 1.4.1.9) is described. Branched-chain L-amino acids were determined with the enzyme by measuring the formation of NADH spectrophotometrically. A linear relationship was established between the absorbance at 340 nm and the concentration of the amino acid (0.02 ~ 0.20 µmol). The determination of branched-chain keto acids (0.01 ~ 0.12 µmol) is based on the measurement of NADH disappeared. This method can be used to assay D-amino acid aminotransferase (EC 2.6.1.21), and is applicable to the assay of other enzymes catalyzing the formation or decomposition of branched-chain L-amino acids and their keto analogs as well.
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