Pomegranate rind has been found to inhibit numerous pathogens, mostly attributed to its tannin fraction. The present study was conducted to investigate the quorum sensing (QS) inhibition effect of tannin-rich fraction from pomegranate rind (TFPR) by using an indicator strain Chromobacterium violaceum. Meanwhile, its effect on biofilm formation and motility of Escherichia coli was evaluated. It was shown that TFPR inhibited QS-regulated violacein pigment production. Biofilm formation and motility of E. coli were also hindered by TFPR. Transcriptional analysis further showed that TFPR repressed expressions of curli genes (csgB and csgD) and various motility genes (fimA, fimH, flhD, motB, qseB, and qseC). Our findings indicated that TFPR has potential application for controlling E. coli contaminations or biofilms in the food industry.