Synthetic microbial community maintains the functional stability of aerobic denitrification under environmental disturbances: Insight into the mechanism of interspecific division of labor

Understanding how synthetic microbial community (SMC) respond to environmental disturbances is the key to realizing SMC engineering applications. Here, dibutyl phthalate (DBP) and levofloxacin (LOFX) were used as environmental disturbances to study their effects on the aerobic denitrification functional stability of SMC composed of Pseudomonas aeruginosa N2 (PA), Acinetobacter baumannii N1(AC) and Aeromonas hydrophila (AH). The results showed that aerobic denitrification efficiency could be maintained at about 93 % under DBP or LOFX disturbance, and interspecific communication was mainly carried out through N-butyryl-L-homoserine lactone (C₄-HSL) and N-(3-oxododecanoyl)-L-homoserine lactone (3OC₁₂-HSL), correspondingly. DBP and LOFX induced the acceleration of tricarboxylic acid (TCA) cycle, which facilitated the energy flux and extracellular polymeric substances (EPS) production, thereby allowing SMC to adapt to disturbances. Under DBP disturbance, DBP stimulated phenazine-1-carboxylic acid production to accelerate electron transfer from the quinone pool to complex III, resulting in an increase in electron transfer activity. Up-regulation of complex I, complex III and heme synthesis genes under LOFX disturbance led to enhanced denitrification enzymes expression and electron transfer efficiency. SMC re-regulated different metabolic pathways to build metabolic networks to maintain normal metabolic activity under different disturbances. Overall, SMC maintained functional stability through the labor division in modulation of interspecific communication, formation of defensive barriers, promotion of energy flux, directional transfer of electron flux, and reconstruction of metabolic networks. DBP stimulated AH and PA to occupy functional dominance, while LOFX induced AC and PA to play a major role. The understanding of the stability mechanism under different environmental disturbances provides valuable guidance for stability maintenance and engineering applications of SMC.