The manometric estimation of L-malate by the malic decarboxylase of Lactobacillu8 arabinosu8 17-5, has been described by Korkes & Ochoa (1948), Ochoa, Salles & Ortiz (1950) and Blanchard, Korkes, del Campillo & Ochoa (1950). As both the bacterial suspensions and the acetone powder extracts used by these workers have fumarase activity, the method measures the sum of malate and fumarate. In the present work, a modification is described whereby L-malate and fumarate are measured separately, and improvements are introduced which are based on the finding that in intact cells the activity of malic decarboxylase is optimal in the presence of glucose, manganous chloride and potassium chloride (Nossal, 1951b). Part of this work has been communicated to the Biochemical Society (Nossal, 1951a). DEVELOPMENT OF THE METHOD Standard conditions for the assay of malic decarboxylase activity, and estimation of L-maliC acid in pure 8olution The main compartment of conical Warburg vessels contained 3-6 ml. malate solution (equivalent to 40jUmol. DLcompound) andO-1 ml. buffer-co-factor solution (1g. glucose, 1 g. MnC1.4Ha0, 14-9 g. KCI and 16-5 ml. 3M-sodium acetate buffer, pH 5-0, in 100 ml.). The side arm contained the bacterial suspension (0-2 ml.) prepared as previously described (Nossal, 1951 b), and 0-1 ml. of buffer-co-factor solution. The gas space contained air. The bath temperature was 35°. These conditions had previously been found to be optimal for the quantitative decarboxylation of malate. The rate of CO2 evolution for 15 min. after mixing the