Graphene-based materials have been extensively researched due to its biocompatibility, unique conjugated structure, and relatively low cost in the realm of biology. Graphene is increasingly being explored for many medical uses, including as DNA-hybridization devices, cancer treatment, and medicine administration. Furthermore, it exhibits substantial potential in several fields of biological applications, such as antibacterial interventions, therapy, and biosensing. The synthesis of GO and rGO was achieved in this work for the first-time using macroalgae, Ulva lactuca as a reducing agent. The antibacterial activity of graphene-based derivatives was evaluated against two different human pathogenic bacteria. Bacterial growth inhibition increased in proportion to the higher concentration. Higher concentration of nanosheets could enhance the cellular enzyme activity. Different scavenging assays are adapted to assess the antioxidant activity of GO and rGO. Significant in-vitro antioxidant activity was observed in a concentration-dependent manner. Furthermore, the antitumor activity of synthesized nanosheets was tested against human breast cancer cells (MCF-7) at various concentrations. The test results show that the graphene derivatives in the MCF-7 cell line exhibit an increase in apoptotic behaviour as sample concentration rises. This study could offer novel insights into the antibacterial, antioxidant, and anticancer activities of the green synthesized graphene-based derivatives.