The textile industry generates significant environmental pollution through its widespread use of synthetic and traditional chemical surfactants in cleaning and finishing processes. Bio-surfactants offer an eco-friendly and sustainable alternative for textile refining. This chapter examines the characteristic features, types, and mechanisms of action of bio-surfactants, focusing on their production through microorganisms and their applications in textile processing. These compounds, characterized by their hydrophilic heads and hydrophobic tails, have diverse applications across the textile industry. The chapter explores how genetic engineering and advanced genome editing tools can enhance bio-surfactant production by leveraging the natural genetic capacity of microorganisms. Applications discussed include soil and dirt removal, fabric treatment, oil removal, and final textile refining. Compared to traditional toxic chemicals that significantly contribute to environmental pollution, bio-surfactants offer both environmental sustainability and economic feasibility, making them an optimal choice for the textile industry.