<p>This preliminary study investigates the feasibility of utilizing Algerian beetroot as a sustainable alternative to commercial Man-de-Rogosa Sharp Agar (MRS) for cultivating lactic acid bacteria. The research explores the optimization of microbial media formulations incorporating beetroot powder and assesses its efficacy in supporting the growth of Lactobacillus plantarum, Lactobacillus fermentum, and Enterococcus durans. Methodologically, vegetal samples were harvested and dried, bacterial strains prepared, and phytochemical analyses conducted. Titratable acidity and sugar content in the beet-based medium were measured, revealing pH 5.5, 13% titratable acidity, and 67&#xa0;g/L sugar content. Results indicate that while beetroot juice alone exhibited limited bacterial growth due to its inadequate nutritional properties, formulations incorporating additives such as magnesium sulfate, ammonium sulfate, and di-potassium phosphate showed improved growth. Particularly, the formulation “BJFA-SM” (with manganese sulfate) demonstrated comparable growth to MRS, with reduced generation times for L. fermentum and E. durans. Additionally, the disappearance of betalain pigment from beetroot-based media indicates successful bacterial growth. Agar plate testing has revealed similar macroscopic characteristics and cell counts to the MRS medium indicating the potential of beetroot-based media as a sustainable substitute for traditional growth media. Finally, the BJFA-MS medium showed superior growth for L. plantarum, E. durans, and L. fermentum compared to MRS, with faster growth rates. However, it exhibited lower acid production efficiency than MRS. This study underscores the viability of beetroot-derived formulations as eco-friendly alternatives for cultivating lactic acid bacteria, with implications for sustainable microbiology research and biotechnological applications.</p>