In this study, the static adsorption behavior and desorption effects of 9 kinds of macroporous resins on natamycin were investigated. The static adsorption kinetics and isothermal adsorption process of the resins were discussed. The in-situ adsorption effects of the macroporous resins on natamycin were also studied in Streptomyces natalensis HW-2 fermentation broth. The results showed that the adsorption and desorption effects of HPD300 and HPD450 resins on natamycin were better, the adsorption rates were 83.47% and 96.14%, respectively. The desorption rates were 95.22% and 85.42%, respectively, and the adsorption capacity was 0.63 and 0.72 mg/g dry resins, respectively. HPD450 was more suitable for the adsorption and analysis of natamycin. According to the Langmuir and Freundlich parameters of isothermal adsorption of HPD450 resin, the adsorption of HPD450 resin conformed to Langmuir equation and was mainly monolayer adsorption. With the Addition of 4 g/30 mL HPD450 resin in 48 hours of fermentation to the fermentation broth for in-situ adsorption, the total yield of natamycin was increased by 66.27%.