Abstract
Essential oil (EO) has significant antifungal activity. However, there is limited information on the mechanism of the synergistic antifungal effect of the effective components of EO against fungi. In the present study, molecular electrostatic potential and molecular docking were used for the first time to investigate the synergistic antifungal mechanism of eugenol and citral small molecule (C(EC)) against Penicillium roqueforti. The results showed that the C(EC) treatment made the activity of β-(1,3)-glucan synthase (GS) and chitin synthase (CS) decreas by 20.2% and 11.1%, respectively, and the contents of which decreased by 85.0% and 27.9%, respectively compared with the control group. Molecular docking revealed that C(EC) small molecules could bind to GS and CS through different amino acid residues, inhibiting their activity and synthesis. The C(EC) can combine with tryptophan, tyrosine, and phenylalanine in the cell membrane, causing damage to the cell membrane. The binding sites between small molecules and amino acids were mainly around the OH group. In addition, C(EC) affected the energy metabolism system and inhibited the glycolysis pathway. Simultaneously, C(EC) treatment reduced the ergosterol content in the cell membrane by 58.2% compared with the control group. Finally, changes in β-galactosidase, metal ion leakage, and relative conductivity confirmed the destruction of the cell membrane, which resulted in the leakage of cell contents. The above results showed that C(EC) can kill P. roqueforti by inhibiting energy metabolism and destroying the integrity of the cell membrane.