Atrazine (ATZ) is a typical environmental endocrine disruptor that is persistent, highly residual, and widely present in various environmental media. Thus, it is of great significance to establish a sensitive and portable detection method for ATZ. In this work, a simple and portable enzymatic biofuel cell-based self-powered sensor (EBFC-SPS) for ATZ is proposed with the ultrasonically dispersed graphene (US-G) serving as the electron transfer promoter for both the enzymatic bioanode and molecularly imprinted cathode. US-G acts as a molecular bridge to stabilize the glucose dehydrogenase (GDH) immobilization and accelerate the electron transfer between GDH and anode, promoting the spontaneous electricity generation by the bioanode and sustainable power supply to the EBFC-SPS while ensuring cathode recognition and electron transfer. The portable detection of ultra-trace ATZ is achieved using a digital multimeter with a wide linear range of 0.5 pM to 100 nM, a low detection limit of 0.37 pM, excellent selectivity and stability in complex actual water samples based on the separation strategy of the bioanode power supply, and molecularly imprinted cathode target recognition. A promising platform for the design of portable sensing strategies with the EBFC-SPS has therefore been provided, which can also be extended to a wider range of environmental pollutants.