Abstract: Preliminary screening was carried out to obtain PI factor structural analogues that can effectively induce natamycin production in the fermentation culture by Streptomyces Chattanovgensis.Box-Behnken experimental design was adopted to build regression model between single factor and natamycin production, according the response surface analysis to determine the optimal conditions. The results indicated that 1, 2-propanediol, glycerol and triethanolamine effectively increased natamycin production.With volume of 0.42, 0.43 and 1 m L for these PI factor structural analogues in 50 m L culture under the condition of120 h, the yield of natamycin was 2.5 g/L, which was 0.60 times higher than that of the control group.It can be seen that a certain amount of PI factor structural analogues can increase the yield of natamycin.