Enhanced Production of 3-Phenyllactic Acid From Novel Non-Axenic Coculture: Adaptive Evolution and Statistical Fermentation Studies

Abstract

This research pivots around screening of idoneous lactic acid bacteria (LAB) from cow milk and subjecting them to adaptive evolution experiments to aid superior growth/robustness necessary for 3-phenyllactic acid (3-PLA) production. Conventional and statistical fermentation studies were conducted at batch scale using a non-axenic coculture of three novel LAB strains: Lactiplantibacillus plantarum str. nov. plantharim, Lactobacillus delbrueckki str. nov. delharim, and Pediococcus pentasaceous str. nov. pentharim. Statistically optimized fermentation using Box Behnken technique resulted in 1225 mg/L 3-PLA production using the growth medium: cheese whey-MRS medium mixture (5:2 ratio), phenylalanine (2.69% w/v), and glucose (9.6% w/v). Statistical optimization of fermentation parameters resulted in a substantial increase (17 times higher) compared to the non-optimized fermentation conditions (72 mg/L). Monad growth kinetics of the cow milk whey (CMW) coculture were calculated and estimated as: mu max = 0.336 h-1, Ks = 11.64 mg/mL, Yx/s = 0.835 mg/g, YP/S = 1.66 mg/g, YX/P = 0.112 mg/mg. The purified 3-PLA (1.93 mg/mL) showed antimicrobial activity with pathogenic bacteria like Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus, with a minimum inhibitory concentration of 12 mg/mL.