Bacterial extracellular polymeric substances (EPS) contribute to the structure, surface charge, and settling properties of activated sludge by assisting in the development of bioflocs. From various soil sources, a total of twenty bacterial isolates were collected. The maximum bacterial isolate was (HS9), which produced (5.8 g/l) after being examined for exopolysaccharide (EPS) synthesis. As a result, EPSs were selected based on Bio-flocculant activity, while HS9 was the most active (86.95 %). The lowest was HS1, while the highest was HS2 (14.55 %). The synthesis and production of EPS are influenced by a variety of variables. As a result, by examining the effects of various factors on the production of EPSs by isolate (HS9), it was discovered that the maximum yield of EPS was (7.9 g/l) and cell dry was (4.1 g/l) when incubation temperature was 40 °C after three days, RPM 120, pH 7, peptone as a nitrogen source, and sucrose (20 g/l) as a carbon source.