DEVELOPMENT OF A NON-DAIRY MEDIUM FOR THE CULTIVATION OF Lacticaseibacillus paracasei.

The demand for healthy food and supplements is constantly increasing, with probiotics being among the most favored products. However, most of the culture media for probiotic production is based on dairy substrates, limiting the options for individuals with milk-related dietary restrictions. In this context, coconut water stands out as a source of nutrients for microbial growth and a low-cost alternative to conventional culture media. Therefore, the objective of this work is to develop a non-dairy culture medium based on coconut water and yeast extract for the probiotic bacteria Lacticaseibacillus paracasei TRA 067308. An experimental design was applied with a factorial 2⁴ matrix and four center points to increase bacterial yield. It was considered as a dependent variable: coconut water (from 25 to 50 g/L), yeast extract (from 5 to 30 g/L), monobasic and dibasic potassium phosphate (from 0 to 2.5 g/L each), and temperature (from 25 to 40 °C). The experiments were conducted in Erlenmeyer flasks containing 100 mL of the designed culture medium sterilized, inoculated with the L. paracasei grown for 20 h at 5% (v/v). The experiments were then incubated on a rotary shaker at 100 rpm for 44 h. Biomass was assessed by measuring optical density (OD), dry weight (DW), and viable cell count (CFU/mL). Statistical analysis was performed using Statistica Six Sigma 8.0 software (StatSoft Inc., USA). The results indicated that a coconut water concentration of 50 g/L and yeast extract at 30 g/L, combined with a temperature of 25 °C and the absence of a buffer system, favored the growth of L. paracasei, reaching an average of 1.5 × 10⁹CFU/mL, 2.37 g/L of DW and 1.472 of OD. This indicates a promising culture media for probiotics, which can be used for the development of non-dairy products after additional optimization and scaling-up studies.