Bacillus subtilis has a wide range of applications in the agriculture, industry, medicine and healthcare sectors due to its wide distribution and harmlessness to the environment. In recent years, Bacillus subtilis has been explored worldwide and applied as a new biological bacteriostatic agent, but the application of antibacterial substances secreted by it is still at a theoretical stage. This paper reviews the classification, mechanism of action, possible future applications and extraction optimization of antibacterial substances from Bacillus subtilis. The production of high-dose and high-purity bacteriostatic substances from Bacillus subtilis is an urgent problem to solved before the wide application of them in food preservation and sterilization technologies. This makes it necessary to find ways to improve the purity and yield of bacteriostatic substances. The review concludes with possible directions for future exploration of bacteriostatic substances.