Abstract: Background and Objective: Fungal endophyte is a good source in producing bioactive compounds possessing antibacterial, antioxidant and anticancer activities. This study was aimed to produce fungal endophyte from Phyllantus niruri and to test its in vitro bioactivity as antibacterial, antioxidant and cancer cytotoxic agent. The molecular mechanism of the fungal activities was then predicted using in silico docking. Materials and Methods: Fungal endophyte from genus Trichaptum sp., isolated from Phyllantus niruri herbs was extracted using ethyl acetate and further fractionated using n-hexane, diethyl ether and ethanol and ethanol 96% through chromatography column ready for testing. The antimicrobial test was carried out using disc diffusion method. The antioxidant was determined using hydrogen peroxide free radical scavenging and reducing power capacity. The cytotoxicity assay against T47D breast cancer cell was carried out using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method. The docking of chemicals reported in the sample was performed using AutoDock Vina. Results: Diethyl ether fraction was the most active fraction against Escherichia coli and T47D cell growth in a non-toxic dose, whereas the best antioxidant activity was performed by ethanol 96% fraction. Assays on the cell cycle modulation and its apoptotic behavior suggested that the T47D was disrupted via p53 mutant, Bcl-2 and NF-κB abrogation. Molecular docking of compounds identified in the fraction suggested that these activities could belong to the maltose binding perisplasmic and oxydosqualene cyclase inhibition in Escherichia coli and T47D, respectively. Conclusion: Fungal endophyte from Phyllantus niruri was successfully isolated as genus Trichaptum sp. and serving as antibacterial, antioxidant and anticancer activities.