Rhamnolipids have recently emerged as promising bioactive molecules due to their novel structures, diverse and versatile biological functions, lower toxicity, higher biodegradability, as well as production from renewable resources. The advantages of rhamnolipids make them attractive targets for research in a wide variety of applications. Especially rhamnolipids are likely to possess potential applications of the future in areas such as biomedicine, therapeutics, and agriculture. The purpose of this mini review is to provide a comprehensive prospective of biosurfactant rhamnolipids as potential antimicrobials, immune modulators, and virulence factors, and anticancer agents in the field of biomedicine and agriculture that may meet the ever-increasing future pharmacological treatment and food safety needs in human health.