This review paper aims to explore the multifaceted roles and applications of microbial surfactants, also known as biosurfactants. These amphiphilic compounds, synthesized by various microorganisms such as Acinetobacter sp., Bacillus sp., Candida antarctica, and Pseudomonas aeruginosa, exhibit both hydrophobic and hydrophilic properties. Through lowering surface tension and interfacial tension, biosurfactants facilitate fluid phase aggregation, imparting organisms with unique physiological and ecological advantages. The synthesis of biosurfactants in bacteria not only contributes to antimicrobial activity but also enhances substrate availability for cellular absorption, particularly in challenging environmental conditions. Categorized based on molecular weight and chemical composition, biosurfactants offer numerous benefits including biodegradability, reduced toxicity, and versatile applications in agriculture, detergent production, biopesticides, and microbial oil enhancement processes. This review synthesizes current knowledge on microbial surfactants, elucidating their diverse roles and promising applications across various industries.