Increased drug resistance has significantly reduced the effectiveness of antibiotics used in the treatment of Propionibacterium acnes. Therefore, there has been a trend toward the development of new antimicrobial agents to circumvent drug resistance. In this study, we isolated and purified a novel bacteriocin, HA2-5, from Bacillus haynesii HA2, which effectively killed P. acnes through membrane disruption at a minimum inhibitory concentration (MIC) of 8 μg/mL. HA2-5 with 2× MIC was able to kill 99.9% of P. acnes within 24 h. HA2-5 shows excellent stability and tolerance to temperature, pH, proteases, chemical reagents, UV radiation, and metal ions, with almost no loss of inhibitory activity after treatment. In addition, the very low hemolytic activity and cytotoxicity suggest that HA2-5 is biosafe. Notably, HA2-5 exhibits preferred antibacterial activity against gram-positive pathogens with an MIC of 16-32 μg/mL. In conclusion, this study shows that bacteriocin HA2-5 has the potential to be used as an alternative to antibiotics for acne treatment.