Live microorganisms with generally recognized as safe (GRAS) status and antagonistic effect against undesired microbes have been extensively applied in different types of food as protective culture. Bacillus could be a promising bio-preservative, due to its ability to efficiently grow in low cost, large-scale fermentation, and to produce pH tolerant bacteriocins which exert broader antimicrobial spectra than those of lactic acid bacteria (LAB). Many research groups have therefore conducted a systematic investigation in all aspects of Bacillus including bacteriocin properties and production. This review presents intensive information of bio-preservative and available commercial bio-protective culture of the Bacillus in comparison with LAB, including (i) bacteriocin classification and properties, (ii) the use of Bacillus as an alternative bio-preservative, (iii) approaches in production of bio-protective culture from Bacillus, and (iv) safety evaluation for a novel bio-protective culture. The results from those studies helped exploring a potential of Bacillus in production and utilization as protective culture.