Rhamnolipids are a class of biosurfactants produced by various bacterial species, particularly Pseudomonas aeruginosa. These molecules have gained significant attention due to their unique properties and potential applications in various industries. Structurally, rhamnolipids consist of one or two rhamnose sugar molecules linked to one or more lipid (fatty acid) chains. This amphiphilic nature, with both hydrophilic and hydrophobic components, gives rhamnolipids their surfactant properties, allowing them to reduce surface and interfacial tensions between different phases. One of the most notable features of rhamnolipids is their ability to enhance the solubilization and biodegradation of hydrophobic compounds, such as oils, hydrocarbons, and other organic pollutants. This property makes them valuable in environmental remediation processes, where they can facilitate the removal of contaminants from soil and water. Additionally, rhamnolipids have shown potential in enhancing oil recovery from reservoirs in the petroleum industry. Rhamnolipids also exhibit antimicrobial and antifungal activities, making them promising candidates for use in various applications, including agriculture, food preservation, and healthcare. Their ability to disrupt cell membranes and inhibit biofilm formation contributes to their efficacy against a wide range of microorganisms. Furthermore, rhamnolipids have found applications in the cosmetic and personal care industries, where they can act as emulsifiers, foaming agents, and solubilizers. Their biodegradability and low toxicity make them attractive alternatives to synthetic surfactants in these products. The production of rhamnolipids can be achieved through microbial fermentation processes using various carbon sources, including renewable resources like agricultural residues and waste materials. This sustainable production method aligns with the growing demand for eco-friendly and bio-based products in various sectors. Despite their promising potential, the large-scale production and commercialization of rhamnolipids have been hindered by challenges related to cost-effective production processes and purification methods. Ongoing research efforts are focused on optimizing production strains, fermentation conditions, and downstream processing to overcome these challenges and facilitate the wider adoption of rhamnolipids in diverse applications.