Mixed matrix membranes blended with graphene-based nanomaterials have great potential in water and wastewater treatment on account of their multiple functionalities. To solve the complicated biofouling problem and diversify the applications of membranes, novel synergistic antibacterial guanidyl-functionalized graphene/polysulfone (GFG/PSF) mixed matrix ultrafiltration membranes were prepared by a non-solvent induced phase separation method. The guanidyl-functionalized graphene nanosheets were achieved by a two-step grafting process consisting of amination and guanidination and exhibited high dispersibility in the casting solution, which showed good compatibility with the polymer matrix. Besides the advantages of partially reduced graphene oxide (GO) nanosheets in creating a stronger interaction with the bacterial cell membrane to destroy the bacteria, the induced bidendate binding between guanidyl groups and phosphate groups on the cell wall can make high sterilization rate even at low concentrations. Different techniques including XRD, FTIR, XPS, SEM, TEM, EDX, contact angle meter, filtration and antibacterial experiments were employed to characterize and investigate the performance of nanosheets and membranes. Compared with pure PSF membrane, the GFG/PSF mixed matrix membranes not only exhibited superior permeability and prominent antifouling property performance toward bovine serum albumin (BSA), but also displayed excellent antimicrobial activity and long-term duration toward Escherichia coli and Staphylococcus aureus. (C) 2019 Elsevier Inc. All rights reserved.