Biosurfactants, characterized by their amphiphilic nature, are synthesized by a diverse array of microorganisms. These microorganisms possess the enzymatic capabilities to decompose complex organic molecules present in agricultural and industrial wastes. Through sophisticated fermentation and biodegradation processes, these substrates are converted into valuable biosurfactants. Agricultural residues, including crop remains, food processing by-products, and industrial effluents, serve as abundant feedstocks for microbial biosurfactant production. The resulting biosurfactants exhibit desirable properties such as biodegradability and low toxicity, making them highly suitable for various industrial applications. However, optimizing the production process remains challenging, necessitating careful consideration of factors such as microbial strain selection, fermentation conditions, and downstream processing. Continuous research is imperative to deepen our understanding of microbial metabolism and enhance bioprocess optimization, thereby promoting greater efficiency and sustainability in the production of biosurfactants from agricultural and industrial wastes. This chapter highlights the crucial role of microorganisms in converting agricultural and industrial wastes into biosurfactants, emphasizing the need for continued research to overcome production challenges and promote sustainable practices.