Dawadawa botso is a nutritious traditional condiment produced by the fermentation of the seeds of Roselle (Hibiscus sabdariffa); it often features in the food of local people in West Africa. Despite the acceptance and importance of this traditionally produced food condiment, it has received little scientific attention. This study was carried out to evaluate the effect of different combinations of fermenting bacteria on the production of dawadawa botso with a view to determining the best starter culture. Isolated fermenting organisms were used to produce dawadawa botso, and their effect on its pH, proximate composition, mineral content and the amino acid profile was determined. The highest pH of 7.22 and lowest of 6.58 was recorded during the fermentation studies. Significant variations (P<0.05) were observed in some of the proximate and mineral compositions of dawadawa botso produced with different starter cultures. Lowest and highest values recorded for lipid was 2.17 and 15.50% respectively, and that of protein and carbohydrate were (15.12 and 27.56%) and (11.04 and 40.72%) respectively. The order of abundance of the mineral content followed the pattern potassium>sodium>phosphorus>magnesium>calcium, showing the most to least in quantity. The major amino acids detected are glutamic acid, aspartic acid and leucine in the unfermented seeds. However, variations were observed after fermentation with the starter cultures. Dawadawa botso produced with all the organisms showed an increase in MSG-like free amino acid classes with 1F organisms showing the lowest value after fermentation. Sweet and bitter free amino acids decreased for the starter combinations used. This finding suggests that the types of fermenting organisms influence the nutritional and organoleptic properties of dawadawa botso.