L-LACTATE DEHYDROGENASE FROM LEAVES OF CAPSELLA-BURSA-PASTORIS (L) MED .1. IDENTIFICATION AND PARTIAL CHARACTERIZATION

By ammonium sulfate fractionation and gel filtration an enzyme preparation which catalyzed NAD+-dependent L-lactate oxidation (10-4 kat kg-1 protein), as well as NADH-dependent pyruvate reduction (10-3 kat kg-1 protein), was obtained from leaves of Capsella bursa-pastoris. This lactate dehydrogenase activity was not due to an unspecific activity of either glycolate oxidase, glycolate dehydrogenase, hydroxypyruvate reductase, alcohol dehydrogenase, or a malate oxidizing enzyme. These enzymes could be separated from the protein displaying lactate dehydrogenase activity by gel filtration and electrophoresis and distinguished from it by their known properties. The enzyme under consideration does not oxidize D-lactate, and reduces pyruvate to L-lactate (the configuration of which was determined using highly specific animal L-lactate dehydrogenase). Based on these results the studied Capsella leaf enzyme is classified as L-lactate dehydrogenase (EC 1.1.1.27). It has a Km value of 0.25 mmol l-1 (pH 7.0, 0.3 mmol l-1 NADH) for pyruvate and of 13 mmol l-1 (pH 7.8, 3 mmol l-1 NAD+) for L-lactate. Lactate dehydrogenase activity was also detected in the leaves of several other plants. © 1979 Springer-Verlag.