A high flux and anti-fouling graphene oxide (GO) nanoparticles embedded in polyacrylonitrile (PAN) nanofiber microfiltration membranes (PANGMs) were fabricated through the facile electrospinning method and were characterized by water treatment applications. The synthesized GO nanoparticles and GO nanoparticles embedded in PAN nanofiber membranes were characterized by FESEM, FTIR, and EDS. SEM images showed that the PANGMs possessed randomly overlaid fibers with a network-like highly porous structure similar to the pristine PAN nanofiber membrane, while agglomeration of GO nanoparticles was observed at high GO concentration. The introduction of GO nanoparticles into the PAN polymeric matrix significantly increased the permeation flux of the resulting membrane in both dead-end and cross-flow filtration systems. A high flux recovery ratio of 96.6% and a low irreversible fouling ratio of 3.4% were obtained at 2% (wt.) GO nanoparticles. More importantly, a high flux recovery ratio of GO nanoparticles embedded in PAN nanofiber membrane was retained after 20 repeated cycles of activated sludge suspension filtration. Therefore, it can speculate that the incorporation of GO nanoparticles into the PAN nanofibers could efficiently improve the anti-fouling ability of membranes which had opened up an alternative for the preparation of high flux and anti-fouling microfiltration membranes in practical water treatment applications such as membrane bioreactors.