Optimization of fermentation conditions for enhanced acetylcholine and biomass production of Lactiplantibacillus plantarum AM2 using the Taguchi approach

This study aimed to optimize the fermentation conditions and medium composition for maximum acetylcholine (ACh) and biomass production by Lactiplantibacillus plantarum AM2 using the Taguchi array design, which enables efficient identification of influential variables through minimal experimental runs. Seven key factors were evaluated: beef extract, peptone, yeast extract, glucose, pH, agitation rate, and inoculation size. The optimization process identified the most significant variables influencing ACh and biomass production, with beef extract and peptone being critical for ACh synthesis, while inoculation size was a critical determinant of biomass yield. The optimal conditions for ACh production were determined as beef extract (11 g/l), peptone (40 g/l), yeast extract (5 g/l), glucose (20 g/l), pH 5.7, no agitation, and 1% (v/v) inoculation size, resulting in a predicted ACh concentration of 490.83 pg/ml and an experimental value of 495.8 pg/ml. For biomass production, the optimal conditions were beef extract (8 g/l), peptone (10 g/l), yeast extract (20 g/l), glucose (35 g/l), pH 6.6, agitation at 150 rpm, and 4% (v/v) inoculation size, yielding a predicted biomass of 20.58 g/l and an experimental value of 21.3 g/l. The optimized conditions significantly improved ACh production (6.32-fold) and biomass production (4.56-fold) compared to basal conditions. These findings highlight the efficiency of the Taguchi approach in enhancing the production of ACh and biomass, providing insights into the functional niche of Lactiplantibacillus plantarum AM2 for potential industrial applications and its use in a symbiotic form.