Recently, there is a growing interest of consumers in natural products thus a large number of natural preservatives have been studied as food additives. Among those natural preservatives, rosemary extract are commonly used by the food industry to extend the shelf life of several products. Nevertheless, the incorporation of the rosemary extract in food matrix is highly limited due to the low water solubility and poor chemical stability of constituents of rosemary extract. In this context, it is needed to find a way that can improve the water solubility to incorporate rosemary extract into aqueous system like foods. Hence, in this study we added surfactants that have a relatively high hydrophilic-liphophilic balance number into rosemary extract solution to increase the water solubility of rosemary extract and then investigated the microbial activities of rosemary extract with surfactants. Tween 20, decaglycerol monooleate, decaglycerol laurate and decaglycerol myristate were used in a range 0.1~4% and rosemary-surfactant solution was prepared by dissolving rosemary in surfactant added phosphate buffer (pH 7) as much as extract can be dissolved. Water solubility of rosemary-surfactant solution were examined by observance of the UV-vis spectra. The antimicrobial effects rosemary-surfactant solution on B.subtilis were also examined by paper disc diffusion method. Rosemary extract showed poor solubility in normal phosphate buffer but its water solubility was highly increased when surfactant was added. This tendency was lasted at all types of surfactant. This is because of amphipathic property of surfactant. Antimicrobial effect to B.subtilis was observed when a relatively lower concentration of surfactants were used, however, not in high concentrations. This result might be attributed to the formation of surfactant micelles containing rosemary extract when a relatively higher concentration of surfactants were used. The information presented may be useful for the development of a new rosemary-loaded delivery systems.