Graphene is a monolayer of tightly packed carbon atoms that attracted tremendous research interest in recent years, owing to its interesting physical, chemical, electrical, mechanical and optical properties and has numerous exciting applications. However, it suffers from low water solubility which limits its biological application. Therefore, the aim of this work is to functionalize the graphene sheets non covalently with various charged surfactants (cationic- Cetrimide, nonanionic-Tween 80, anionic-Sodium dodecyl sulfate) to get a highly stable conjugate soluble in water and to improve the antibacterial activity of this nano-derivative. The morphology and the size of the functionalized graphene were determined by atomic force microscopy. Moreover, the functionalized graphene was quantified by thermogravimetric analysis obtaining the maximum functionalization in the case of citrimide functionalized graphene. The antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa has been studied. The results indicate that only the functionalized graphene with citrimide has significant antibacterial activity against the two strains of bacteria S. aureus and P. aeruginosa with MIC 0.98, 7.81 mg/L respectively.