Shigella flexneri (S. flexneri), a major intestinal pathogen, is a global public health concern. The biofilms formed by S. flexneri threaten environmental safety, since they could promote the danger of environmental contamination and strengthen the disease-causing properties of bacteria. Epigallocatechin gallate (EGCG) is an important catechin in tea, which has a high antibacterial activity. However, its antibacterial mechanism is still unclear. This research aims to quantify the antibacterial function and investigate the possible mechanism of EGCG inhibition of S. flexneri. The minimum inhibitory concentration (MIC) of EGCG against planktonic S. flexneri in the investigation was measured to be 400 μg/mL. Besides, SDS-PAGE and field emission scanning electron microscopy showed that EGCG interfered with protein synthesis and changed bacteria morphology. Through controlling the expression of the mdoH gene, EGCG was found to be able to prevent an S. flexneri biofilm extracellular polysaccharide from forming, according to experiments utilizing the real-time PCR test. Additional research revealed that EGCG might stimulate the response of S. flexneri to oxidative stress and prevent bacterial growth. These findings suggest that EGCG, a natural compound, may play a substantial role in S. flexneri growth inhibition.