Enzymic Assays of Amino Acids and Keto Acids

  • B. D. Sanwal
Part of the Modern Methods of Plant Analysis / Moderne Methoden der Pflanzenanalyse book series (PFLANZENANAL., volume 6)


An enzyme can be used for the estimation of a given metabolite only if it fulfills the following criteria: 1. The enzyme is specific for the metabolite in question. 2. The reaction proceeds to completion and, 3. one of the products of the reaction can be easily measured. In many cases, however, all the criteria are not easily fulfilled. In the spectrophotometric measurement of aspartic acid, for instance, use is made of the glutamate-oxalacetate transaminase which converts aspartate with α-ketoglutarate to glutamate and oxalacetate. This reaction, however, does not go to completion; equilibrium is established when the concentration of oxalacetate reaches 0.15 mole per mole of aspartate in solution (Pfleiderer, Gruber and Wieland, 1955). The reaction can be brought to completion if the oxalacetic acid is continuously removed with the help of DPNH and 1-malic dehydrogenase. Many such coupled reactions are used in the determination of the metabolites.


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© Springer-Verlag oHG. Berlin · Göttingen · Heidelberg 1963

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  • B. D. Sanwal

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