Malic enzymes are a class of oxidative decarboxylases that catalyze the oxidative decarboxylation of malate to pyruvate and carbon dioxide, with concomitant reduction of NAD(P)(+) to NAD(P)H. The NADP(+)-dependent malic enzyme in oleaginous fungi plays a key role in fatty acid biosynthesis. In this study, the malic enzyme-encoding complementary DNA (cDNA) (malE1) from the oleaginous fungus Mortierella alpina was cloned and expressed in Escherichia coli BL21 (DE3). The recombinant protein (MaME) was purified using Ni-NTA affinity chromatography. The purified enzyme used NADP(+) as the cofactor. The K (m) values for l-malate and NADP(+) were 2.19 +/- 0.01 and 0.38 +/- 0.02 mM, respectively, while the V (max) values were 147 +/- 2 and 302 +/- 14 U/mg, respectively, at the optimal condition of pH 7.5 and 33 A degrees C. MaME is active in the presence of Mn2+, Mg2+, Co2+, Ni2+, and low concentrations of Zn2+ rather than Ca2+, Cu2+, or high concentrations of Zn2+. Oxaloacetic acid and glyoxylate inhibited the MaME activity by competing with malate, and their K (i) values were 0.08 and 0.6 mM, respectively.