Polysulfone (PS) membranes blended with different loadings of arabic gum (AG) were synthesized using phase inversion method and the antibacterial properties of the synthesized membranes were tested using a number Gram-negative (Escherichia coli, Klebsiella pneumonia and Pseudomonas aeruginosa) and Gram-positive (Staphylococcus aureus) bacterial species. It was shown that AG addition to the dope polymer solutions essentially changed porous structure, hydrophilicity and zeta potential of the cast PS/AG membranes. These changes were due to the amphiphilic properties of AG macromolecules that contained negatively charged hydrophilic residues. A pronounced decrease in bacterial attachment was seen in the field emission scanning electron microscopy (FESEM) images for PS/AG membrane samples compared to both commercial (Microdyn-Nadir) and bare PS (without AG) membranes. AG loading dependent trend was observed where the prevention of bacterial colonization on the membrane surface was strongest at the highest (7 wt. %) AG loading in the casting solution. Possible mechanisms for the prevention of bacterial colonization were discussed. Significantly, the inhibition of bacterial attachment and growth on PS/AG membranes was observed for both Gram-positive and Gram-negative bacterial models, rendering these novel membranes with strong biofouling resistance attractive for water treatment applications.