Batch-mode degradation of high-strength phenolic pollutants by Pseudomonas aeruginosa strain STV1713 immobilized on single and hybrid matrices.

Controlled environments are pivotal in all bioconversion processes, influencing the efficacy of biocatalysts. In this study, we designed a batch bioreactor system with a packed immobilization column and a decontamination chamber to enhance phenol and 2,4-dichlorophenol degradation using the hyper-tolerant bacterium Pseudomonas aeruginosa STV1713. When free cells were employed to degrade phenol and 2,4-DCP at a concentration of 1000 mg/L, the cells completely removed the pollutants within 28 h and 66 h, respectively. Simultaneous reductions in chemical oxygen demand and biological oxygen demand were observed (phenol: 30.21 mg/L/h and 16.92 mg/L/h, respectively; 2,4-dichlorophenol: 12.85 mg/L/h and 7.21 mg/L/h, respectively). After assessing the degradation capabilities, the bacterium was immobilized on various matrices (sodium alginate, alginate-chitosan-alginate and polyvinyl alcohol-alginate) to enhance pollutant removal. Hybrid immobilized cells exhibited greater tolerance and degradation capabilities than those immobilized in a single matrix. Among them, polyvinyl alcohol-alginate immobilized cells displayed the highest degradation capacities (up to 2000 mg/L for phenol and 2500 mg/L for 2,4-dichlorophenol). Morphological analysis of the immobilized cells revealed enhanced cell preservation in hybrid matrices. Furthermore, the elucidation of the metabolic pathway through the catechol dioxygenase enzyme assay indicated higher activity of the catechol 1,2-dioxygenase enzyme, suggesting that the bacterium employed an ortho-degradation mechanism for pollutant removal. Additionally, enzyme zymography confirmed the presence of catechol 1,2-dioxygenase, with the molecular weight of the enzyme determined as 245 kDa.

Revealing the degradation mechanisms of the hyper-tolerant bacterium Pseudomonas aeruginosa STV1713 under high phenol and 2,4-DCP-induced stress conditions through RNA-seq analysis.

The ability of bacteria to efficiently remove phenolic pollutants depends on their genetic makeup and environmental conditions. This study examined a novel strain, Pseudomonas aeruginosa STV1713, for degrading higher concentrations of phenol and 2,4-dichlorophenol. After optimization, a combination of degradation parameters, such as pH (7.0), temperature (32.5 °C), and ammonium nitrate concentration (0.7 g/L), was found to reduce degradation time while promoting cell growth. Under these optimal conditions, the bacterium effectively degraded up to 2000 mg/L of phenol and 1400 mg/L of 2,4-dichlorophenol, while maximum tolerance was observed till 2100 mg/L and 1500 mg/L, respectively. Metabolic profiling identified crucial metabolites in the ortho-degradation pathway during pollutant removal. Additionally, transcriptome analysis revealed that P. aeruginosa STV1713 utilizes different branches of the beta ketoadipate pathway for phenol and 2,4-DCP removal. Moreover, under high pollutant stress, the bacterium survived through differential gene expression in ribosome biogenesis, chemotaxis, membrane transport, and other pathways.