Microorganisms producing extracellular polymeric substances (EPS) are of great potential in numerous environmental applications. The present study explores the production and properties of extracellular polymeric substances (EPS) from Amphora coffeiformis diatom strain and their potential applications in environmental remediation. EPS were composed of a complex mixture of polysaccharides, proteins, humic substances and nucleic acids, with polyanionic characteristics as revealed by FTIR, Raman and zeta potential analyses. EPS showed high flocculation efficiency against kaolin clay at low dosages (5 mg/L) through a charge neutralization mechanism involving both polysaccharides and proteins. EPS also exhibited strong emulsification activity for various nonpolar substrates, mainly olive oil, corn oil, soybean oil, essence and diesel, with emulsification indexes above 80%. The emulsions were stable for 72 h under different NaCl concentrations (1-10% w/v). Moreover, EPS demonstrated remarkable adsorption capacity for lead, reaching a maximum of 1699.33 ± 89.61 mg/g under optimized conditions using Box-Behnken design. The adsorption mechanism involved multiple functional groups such as hydroxyl, carbonyl, carboxyl, phosphoric and sulfhydryl. Therefore, EPS from A. coffeiformis are a promising candidate for restoring environments contaminated by heavy metals.