Optimization of industrial (3000 L) production of Bacillus subtilis CW-S and its novel application for minituber and industrial-grade potato cultivation

Abstract

A commercial plant probiotic product was developed employing Bacillus subtilis CW-S in submerged fermentation. The effects of molasses and urea on cell growth were investigated with the goal of low-cost manufacturing. Plackett-Burman and Central-Composite Design (CCD) were utilized to optimize production parameters to maximize productivity. The stability of the formulated product and its efficacy in cultivating minituber in aeroponics and industrial-grade potatoes in the field were assessed. The results showed that the medium BS10 (molasses and urea) produced satisfactory cell density (7.19 x 10(8) CFU/mL) as compared to the control (1.51 x 10(7) CFU/mL) and BS1-BS9 (expensive) media (1.84 x 10(7)-1.37 x 10(9) CFU/mL). According to validated CCD results, optimized parameters fitted well in pilot (300 L; 2.05 x 10(9) CFU/mL) and industrial (3000 L; 2.01 x 10(9) CFU/mL) bioreactors, resulting in a two-fold increase in cell concentration over laboratory (9.84 x 10(8) CFU/mL) bioreactors. In aeroponics, CW-S produced excellent results, with a significant increase in the quantity and weight of minitubers and the survival rate of transplanted plantlets. In a field test, the yield of industrial-grade (> 55 mm) potatoes was increased with a reduction in fertilizer dose. Overall, the findings suggest that CW-S can be produced commercially utilizing the newly developed media and optimized conditions, making plant probiotics more cost-effective and accessible to farmers for crop cultivation, particularly in aeroponic minituber and industrial-grade potato production.