A quantitative method for assaying bioemulsifiers in culture broth was developed and applied to cultivation of Pseudomonas aeruginosa YPJ80. SED(Standard Emulsification Dilution), an indirect measure of bioemulsifier concentration, was proposed. Production of bioemulsifier and rhamnolipid reached their maximum simultaneously. However, the bioemulsifier/rhamnolipid ratio decreased with cultivation time. This indicates the presence of another bioemulsifier other than rhamnolipid. The bioemulsifier seems to be protein-like activator which showed emulsification activity in addition to rhamnolipid.