Biofilms are composed primarily of microbial cells and EPS. Biochemical analysis of EPS extracted from different coupons was carried out the protein content was found to be more abundant than carbohydrate in both enriched and normal biofilms during the exposure period. The protein, carbohydrate, calcium and magnesium concentrations in the EPS of biofilm grown on stone coupons were higher than the EPS of biofilm grown on PVC, wood and glass coupons. Nutrient boosted the biofilm cells growing in enriched medium which resulted in more EPS production.