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Growth requirements for maximum production of pediocin Iz3.13 by Pediococcus pentosaceus Iz3.13 were studied. Physical and chemical parameters that enhanced pediocin production included incubation in static broth at 30℃, with an initial pH of 4.5. Optimum nutritional conditions that enhanced pediocin Iz3.13 production were 3.5% Trypticase peptone, 3.5% Phytone peptone or 5.0% Proteose peptone No.3 as nitrogen sources, and glucose or maltose as carbon sources. Supplementing growth media with beef extract and MgCl2 also enhanced production. This study proposes an optimized growth medium containing 5.0% Proteose peptone No.3, 3.5% Phytone peptone, 2.5% glucose, 1.5% MgCl2, 3.0% beef extract and 0.25% K2HPO4. This medium significantly enhanced pediocin Iz3.13 production (5.1 fold increase) as compared to a basal medium.