Enantioselective toxicity and mechanism of chiral fungicide penflufen based on experiments and computational chemistry

Penflufen fungicide is widely used as a racemate, which has potential ecological risks to aquatic organisms, while its enantioselective toxicity data is limited. This study aimed to differentiate the enantioselective toxicity difference of penflufen enantiomers, and illuminate the enantioselective mechanism from the insight of enantiomer-protein specific binding. The semipreparative separation and absolute configuration of penflufen enantiomers were conducted. The acute toxicity of S-(+)-penflufen was 54 times higher than R-(-)-penflufen to Danio rerio, and the coexistence of R-(-)-penflufen could increase the exposure risk of S-(+)-penflufen. For chronic toxicity, after low-dose long-term exposure, rac-penflufen and S-(+)-penflufen inducted more serious oxidative stress than R-(-)-penflufen in D. rerio, and inhibited the succinate dehydrogenase (SDH) activity significantly. For target phytopathogen, the toxicity difference of S-(+)-penflufen and R-(-)-penflufen was up to 148 times for Rhizoctonia solani. Based on the toxic unit analysis, the toxic interactions of antagonistic effect and concentration addition were found between penflufen enantiomers, indicating the coexistence of R-(-)-penflufen could increase overuse and environmental risks. Computational chemistry was used to illuminate the enantioselectivity mechanism, and the lower binding energy between the active site of SDH and S-(+)-penflufen contributed to the higher toxicity. The higher target toxicity might be due to the hydrophobic pocket of CybL in R. solani was more benefited to S-(+)-penflufen binding SDH than Botrytis cinerea. These results could be helpful for further understanding the potential risk of chiral penflufen in the environment, demonstrating the importance of understanding the enantioselective difference of chiral pesticides, and providing a new insight for analyzing the enantioselective mechanism. [Display omitted] •Absolute configuration was confirmed using computational method of quantum mechanics.•Toxicity difference of S-penflufen and R-penflufen was up to 54 times for D. rerio.•S-penflufen was148 times more toxic than R-penflufen for target R. solani.•Enantiomer-protein specific binding was used to analyzing enantioselective mechanism.•Compared to R-penflufen, the stronger binding of S with SDH led to higher toxicity.