The production conditions of kocurin by the Kocuria strain F-276,345 were analyzed with a time course study monitoring the growth and kocurin production in the medium R358 during 4 days. Given that our preliminary experiments using standard seed ratios of 5% had shown a rapid exponential growth and production of the compound in concentrations of 0.2 μg/mL within the first 24 h of cultivation, the experiment was established using diluted seed conditions ensuring initial optical densities as low as OD600 0.015. Using this approach we have confirmed that whereas the strain reached the stationary phase within the first 48 h, kocurin production, estimated as the intensity of the ion M + 2Na+ with m/z = 780, was initiated during the exponential growth reaching its maximal production 24 h after the establishment of the stationary phase to fall afterwards as the incubation proceeds up to 4 days