The effects of the addition of short-chain fatty acids and lower alcohols on the production of natamycin from Streptomyces natalensis F4-245 was investigated, and propanol was found to be the most effective additive. Under the optimal condition of propanol addition, the maximal natamycin titer reached 10.38 g/l, which was 17 % higher than that of the control. Metabolites analysis showed the concentrations of amino acids and acetyl-CoA were enhanced while those of organic acids in tricarboxylic acid (TCA) cycle were reduced. This work demonstrates that the addition of propanol is an effective strategy to increase natamycin yield through regulating metabolite node and pools of precursors.