Bacteriocin, is an antimicrobial peptide produced by the bacteria itself, including the lactic acid bacteria. Some of them are also known for spermicidal activities. Thus, it is potential to be developed as a spermicidal agent to prevent sexual transmitted disease. Bacteriocin Nisin and Subtilosin, produced by Lactococcus lactis and Bacillus subtilis respectively, have been reported to exhibit this activity. Previous study on Weissella confusa MBF8-1 bacteriocin activity reported that its bacteriocin like inhibitory substance (BLIS) showed an antibacterial activity against several bacteria. This study aimed to investigate the spermicidal activity of its BLIS, as well as the recombinant and synthetic Bacteriocin peptides Bac1, Bac2, Bac3. These three bacteriocin peptides were reported to be existed in W. confusa MBF8-1 previously as carried by the plasmid pWcMBF8-1. The screening for spermicidal activity was conducted by using the modified Sander-Cramer assay and the statistical analysis has been done by using SPSS. Recombinant peptide and synthetic peptide of Bac1, Bac2 and Bac3, either as single or in combination were tested. Result showed that the combination of Bac1, Bac2 and Bac3 in synthetic peptide form showed the highest spermicidal activity compares to BLIS, recombinant form, and other synthetic peptide combination, with the increase of motility score by 38.25 points in 30 seconds contact, and 39 points in 5 minutes contact. However, none of BLIS, recombinant peptide and synthetic peptide of Bac1, Bac2 and Bac3, either as single or in combination, showed strong spermicidal activity as Nisin did.