In the last recent years, bacteriocins produced by lactic acid bacteria (LAB) have received a considerable attention to be used as biopreservatives in different fields of food industry. LAB was isolated from Sudanese fresh sausages, intestines of different animals, saliva, cheese and cucumber in de Man Regosa and Sharp medium. In this study the antimicrobial effect of LAB producing-bacteriocins against Staphylococcus aureus ATCC 2818, Bacillus cereus, and Escherichia coli ATCC 29522 was studied. The agar well diffusion assay (AWDA) was followed to determine the antimicrobial effect. Characterization of bacteriocin produced by LAB was conducted by exposing bacteriocins to different heat treatment at 40, 60, 80, 100, and 121OC for 0, 30, 60 or 90 minutes and after a week in a water bath set up at 37 OC, and adjusted to different pH levels at 2, 4, 6, 8, 10 and 12. Sensitivity to enzymes was assayed by treatment with α-amylase and the proteolytic enzyme pepsin. Only 16 LAB isolates of 30 produced clear inhibition zones. Only 3 LAB isolates of 16 exhibited the strongest performance against the indicator organisms. These LAB isolates were identified as Enterococcus faecalis, Pediococcus pentosaceus, and Lactobacillus murinus. Characterization of bacteriocins obtained revealed that these substances were found to be stable after heat treatment, at low pH and at pH 6-8. The proteinaceous nature of bacteriocins was completely lost after treatment with the proteolytic enzyme pepsin. Sudanese products of animal sources may be a rich source for bacteriocins-producing LAB which can be used as bio-preservatives