A simple, sensitive agar plate method was used to screen lipase activity from 1229 selected cultures, including 508 bacteria, 479 yeasts, 230 actinomycetes and 12 fungi, covering many genera and species. About 25% of the cultures tested are lipase-positive. These lipase-positive strains were further classified into three categories according to their enzyme activity: good, moderate and weak lipase producers for those having orange fluorescent halo zones greater than 10 mm, 7.5 mm or 5 mm diameters, respectively. The good lipase producers have the potential to be developed for industrial enzymes. The positional, fatty acid or enantiospecificity of each individual lipase requires further investigation. The data presented here are important for the discovery of new lipases.