In this study, we developed and optimized a growth media by evaluating various nitrogen sources for the cultivation of Lactobacillus bulgaricus, a probiotic and an important dairy starter culture. We modified the composition of deMan, Rogosa and Sharpe (MRS) culture media and substituted the nitrogen content with alternative nitrogen sources X-Seed KAT, X-Seed Carbo Max and X-Seed Nucleo Max in various blends of 5 g/l and 10 g/l respectively. Results showed that bacterial growth was significantly higher when the nitrogen source blend KCMax (10/10) was used. The optical density (OD610 nm) of the Lactobacillus bulgaricus strains were higher (1.34 and 1.79) in the KCMax (10/10) medium than in the MRS medium (0.89 and 1.42) (P < 0.05). There was no significant difference in the bacterial counts for both the MRS medium and the KCMax (10/10) medium, and all bacterial counts were estimated at 8 log CFU/ml. The buffering capacity of KCMax (10/10) was also tested and supplemented with l-histidine and was significantly different (P < 0.05) than that of the MRS control medium. Calcium supplemented in the KCMax (10/10) also served as a cryoprotectant for the cells during freezing and freeze-drying. Bacterial counts of the recovered calcium-treated freeze-dried cells were statistically significant (P < 0.05). We hypothesized that alternative nitrogen sources such as selected yeast extracts from the X-Seed brand of complex nitrogen sources could efficiently support the viability of Lb. bulgaricus. Our results thus suggested the growth of Lb. bulgaricus was efficiently supported by the X-Seed KAT, X-Seed Nucleo Max and X-Seed Carbo Max nitrogen sources. Consequently, these alternative nitrogen sources could potentially be recommended for dairy starter culture fermentations.