Biofouling is an important biological constrain in the water treatment process, and the control or management strategies using green principles have gained recent attention. Antibiofouling agents based on the biological source are now extensively studied due to their high efficacy and are environmentally friendly. In this present study, Poly Acrylo Nitrile (PAN)/Ulvan hollow fibre membranes of four different modules were fabricated for water treatment studies by testing against B. subtilis and E. coli along with separation efficiency studies on proteins such as albumin, pepsin, and clay. Ulvan (Ulv), green seaweed sulfated polysaccharide extracted from Ulva fasciata, was coated on PAN hollow fibres, fabricated using a wet-spinning process. Ulvan was dip-coated on membrane surface followed by cross-linking and resultant changes in terms of performance and morphology. PAN/Ulv hollow fibre membranes were examined for the pure water flux and protein separation analysis to analyse the membrane efficiency. SEM was used to analyse the membrane structure and ATR- FTIR for the determination of functional groups. Ulvan coated (310 C) hollow fibre membrane showed better performance than the other three membranes with a flux of 398.1 L m−2 h−1. Higher sample concentration of suspended solids paved simpler route for enhanced COD sequestration efficacy. Pepsin, albumin, and clay particles were rejected by the 310C Ulvan adorned membranes at rates greater than 80%. By incrementing suspended particles beyond 3200 mg/L, the greatest power recovery was reduced that portrays an adverse influence of bio-fouling process on membrane operation. Experimental results demonstrated that synthesised 310 C membrane possessed better separation performance and antifouling characteristics for aquatic water systems.