l-Ornithinedecarboxylase(ODC)istherate-limitingenzymeofdenovopolyaminesynthesis in humans and fungi. Elevated levels of polyamine by over-induction of ODC activity in response to tumor-promoting factors has been frequently reported. Since ODC from fungi and human have thesamemolecularpropertiesandregulatorymechanisms,thus,fungalODChasbeenusedasmodel enzyme in the preliminary studies. Thus, the aim of this work was to purify ODC from fungi, and assess its kinetics of inhibition towards various compounds. Forty fungal isolates were screened for ODC production, twenty fungal isolates have the higher potency to grow on L-ornithine as sole nitrogensource. AspergillusterreuswasthemostpotentODCproducer(2.1µmol/mg/min),followedby PenicilliumcrustosumandFusariumfujikuori. Theseisolatesweremolecularlyidentifiedbasedontheir ITSsequences,whichhavebeendepositedintheNCBIdatabaseunderaccessionnumbersMH156195, MH155304 and MH152411, respectively. ODC was purified and characterized from A. terreus using SDS-PAGE,showingawholemoleculemassof~110kDaanda50kDasubunitstructurerevealingits homodimericidentity. Theenzymehadamaximumactivityat37◦C,pH7.4–7.8andthermalstability for20hat37◦C,and90daysstoragestabilityat4◦C.A.terreusODChadamaximumaffinity(Km)for l-ornithine, l-lysine and l-arginine (0.95, 1.34 and 1.4 mM) and catalytic efficiency (kcat/Km) (4.6, 2.83, 2.46×10−5 mM−1·s−1). The enzyme activity was strongly inhibited by DFMO (0.02 µg/mL),curcumin (IC50 0.04 µg/mL), propargylglycine (20.9 µg/mL) and hydroxylamine (32.9 µg/mL). These results emphasize the strong inhibitory effect of curcumin on ODC activity and subsequent polyamine synthesis. Further molecular dynamic studies to elucidate the mechanistics of ODC inhibition by curcumin are ongoing.