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Conceptual Error in Determination of NAD⁺-Malic Enzyme in Extracts Containing NAD⁺-Malic Dehydrogenase ¹

Plant Biology Program, Department of Biology (Box 1137), Washington University, St. Louis, MO 63130

The typical procedure for determining NAD⁺-malic enzyme (EC 1.1.1.39) is to calculate the enzyme rate to be A₃₄₀/ time after the endogenous NAD⁺-malic dehydrogenase (EC 1.1.1.37) catalyzed reaction has reached equilibrium. This ignores the equilibrium shift of oxaloacetic acid and NADH during the course of the NAD⁺-malic enzyme reaction and causes an error that varies depending on the reagent [malate], [NAD⁺], pH and final [NADH]. For a A₃₄₀ of 0.02, the error is about 80% and for a A₃₄₀ of 0.30, 20%. We develop this argument, give supportive data and present a simple method to circumvent the error.

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