Nonactin is the parent compound of a group of ionophore antibiotics, that known as the macrotetrolides. In previous report, in th course of screening superoxide radical-generating compounds from microbial sources, we first screened Streptomyces viridochromogenes JM-4151 that produces nonactin. It was proved that nonactin is superoxide radical-producing compound. In present study, we examined the optimal culture conditions of nonacin. Th optimal culture conditions for nonactin production were as follows: 1% soluble starch, 1% yeast extract, 0.2% ammonium nitrate, 0.06% magnesium sulfate, 0.2% calcium carbonate, initial pH 7.0 at 28$^{circ}C$ for 96 h. The highest nonactin production was achieved in the production medium of initial pH7.0 at 28$^{circ}C$ for 96h. The threshold level of dissolved oxygen was found to be above 33.2% at 28$^{circ}C$ when 1% soluble starch was used as a carbon source. These results suggest that S. viridochromogenes JM-4151 might be a possible strain for industrial nonactin producer.