Due to its abundance, low cost and sustainability, sweet potato, a widely cultivated tuberous root rich in carbohydrates and essential nutrients, presents an intriguing prospect as a potential growth medium for Lactobacillus.sp. In the present study, we investigated the feasibility and efficacy of sweet potato-based medium for supporting the growth of Lactobacillus.sp bacteria. The sweet potato medium was supplemented with 2, 8, or 14 g/L of plant- based nitrogen sources (X-Seed Nucleo Max, X-Seed KAT, and NuCel 780 MG) to form SPM1, SPM2, and SPM3 respectively. Seven Lactobacillus.sp strains were used in this study, and a Lactobacilli MRS growth medium was used as a control. The growth pattern of tested Lactobacillus.sp strains grown in SPM2 and SPM3 was similar or higher than that in MRS. The population of Lactobacillus.sp strains reached an average of 7.94 ± 0.11, 8.38 ± 0.11, and 8.57 ± 0.12 at 12 h of incubation for MRS, SPM2, and SPM3, respectively. The buffering capacity of SPM2 and SPM3 was significantly (p < 0.05) higher than that in MRS, and the growth in SPM1 was lower than that of MRS. These findings demonstrate that SPM2 is suitable for the growth of Lactobacillus.sp strains and could be used as an alternative medium.