The present study aimed to characterize lactic acid bacteria involved in the different processing steps of tchapalo, a traditional Ivoirian beverage, for their potential application as starter cultures in food and beverages. Lactic acid bacteria (LAB) were therefore isolated and enumerated at different steps of the process on MRS and BEA agars. Of the 465 isolates, 27 produced bacteriocins that inhibit Lactobacillus delbrueckii F/31 strain. Of those, two also inhibited Listeria innocua ATCC 33090, while two others displayed inhibitory activity against L.innocua ATCC 33090, E. faecalis CIP 105042, E. faecalis ATCC 29212, Streptococcus sp. clinical LNSP, E. faecalis CIP 105042 and E. faecium ATCC 51558. The dominant species involved in tchapalo LAB fermentation, as determined by 16S rRNA gene sequencing, were Lactobacillus fermentum (64%), followed by Pediococcus acidilactici (14%). Two strains representing the two dominant species, L. fermentum S6 and P. acidilactici S7, and two potential bacteriocin producers, Weissella confusa AB3E41 and Enterococcus faecium AT1E22, were selected for further characterization. First, genome analysis showed that these strains do not display potential harmful genes such as pathogenic factors or transmissible antibiotic resistance genes. Furthermore, phylogenetic analyses were performed to assess evidence of eventual links to groups of strains with particular properties. They revealed that (i) L. fermentum S6 and P. acidilactici S7 are closely related to strains that ferment plants, (ii) E. faecium AT1E22 belongs to the environmental clade B of E. faecium, while W. confusa is quite similar to other strains also isolated from plant fermentations. Further genome analysis showed that E. faecium AT1E22 contains the Enterocin P gene probably carried by a megaplasmid, whereas no evidence of a bacteriocin gene was found in W. confusa AB3E41. The metabolic and the first step of the probiotic potentials of the different strains were analyzed. Lactobacillus fermentum S6 and P. acidilactici S7 are good candidates to develop starter cultures, and E. faecium AT1E22 should be further tested to confirm its potential as a probiotic strain in the production of sorghum wort.