Lactic acid bacteria (LAB) exopolysaccharides (EPSs) have garnered significant scientific interest due to their multifaceted roles in food technology and health promotion. This comprehensive review systematically examines the structural classification of LAB EPSs, emphasizing distinctions between homo-and heteropolysaccharides, as well as the influence of substituent groups (e. g., acetyl, phosphate) on their physicochemical and bioactive properties. Advanced isolation methodologies, including ethanol precipitation and ultrafiltration, coupled with characterization techniques such as nuclear magnetic resonance (NMR) spectroscopy and atomic force microscopy (AFM), are vital for deciphering their chemical and physical characteristics. The biosynthesis pathway, governed by eps operons and modulated by environmental factors (e.g., carbon sources, Ca²⁺), are discussed as targets for genetic engineering to enhance yield and functionality. Functionally, LAB EPSs display antioxidant, immunomodulatory, anti-tumor, anti-viral, and anti-biofilm activities, with demonstrated applications as natural additives in the food industry, prebiotics, and drug delivery systems. Despite their potential, challenges such as cost-effective production and regulatory hurdles persist. Future research should prioritize the elucidation of molecular mechanisms, clinical validation of health claims, and sustainable bioprocessing innovations to fully harness the transformative potential of LAB EPSs across food, pharmaceutical, and agricultural industries.