Unveiling the hydrolase Oph2876 mediated chlorpyrifos degradation mechanism in Pseudomonas nitroreducens and its potential for environmental bioremediation.

Chlorpyrifos contamination is a currently on-going issue with significant environmental impacts. As such, rapid and effective techniques that remove chlorpyrifos from the environment are urgently required. Here, a strain of Pseudomonas nitroreducens W-7 exhibited exceptional degradation ability towards both chlorpyrifos and its major metabolite 3,5,6-trichloro-2-pyridinol (TCP). W-7 can effectively reduce the toxicity of chlorpyrifos and TCP towards a variety of sensitive organisms through its superior degradation capacity. W-7 demonstrated efficient soil bioremediation by removing over 50 % of chlorpyrifos (25 mg/kg) from both sterile and non-sterile soils within 5 days, with significantly reduced half-lives. Additionally, 16S rDNA high-throughput sequencing of the soil revealed that the introduction of W-7 had no significant impact on the soil microbial community. A pivotal hydrolase Oph2876 containing conserved motif (HxHxDH) and a bimetallic catalytic center was identified from W-7. Oph2876 was a heat- and alkali-resistant enzyme with low sequence similarity (< 44 %) with other reported organophosphorus hydrolases, with a better substrate affinity for hydrolysis of chlorpyrifos to TCP. The molecular docking and site-directed mutagenesis studies indicated that the amino acid residues Asp235, His214, and His282, which were associated with the conserved sequence "HxHxDH", were crucial for the activity of Oph2876. These findings contribute to a better understanding of the biodegradation mechanism of chlorpyrifos and present useful agents for the development of effective chlorpyrifos bioremediation strategies.