The individual and combined inhibitory activities of propolis extract (PE) and Hermetia illucens larval proteins (Black soldier fly larvae proteins) (BSFLP) against Aspergillus niger were investigated. The antifungal effects of PE, BSFLP and Synergy of PE and BSFLP (SPB) against Aspergillus niger were determined by mycelial growth, spore germination rate and hyphal elongation. Membrane integrity was measured by assessing changes in extracellular proteins, reducing sugars, and electrical conductivity. Intracellular MDA, H2O2, ergosterol, and mannan contents were also evaluated. SEM was used to observe the morphology of the Aspergillus niger mycelium. Additionally, AutoDock Tools software and multi-ligand docking methods were used to study the synergistic interactions of ligand combinations on the receptor. The results showed that the combination of PE and BSFLP had excellent effects. During the treatment of antifungal agents, the membrane permeability changed and a large amount of content leaked, while control check (CK) remained almost unchanged. Ergosterol content was found to increase, while mannan content decreased significantly, leading to cell death. In addition, SPB can cause abnormal accumulation of MDA and H2O2, leading to oxidative stress in cells. SEM results showed that morphology of Aspergillus niger treated with SPB was severe wrinkling, distortion and even breakage. Multi-ligand docking results showed that the binding affinity of receptor 1KS5 to 14 complex ligands increased, as indicated by the decrease in docking energy values. These results provide scientific basis for further study on the application of SPB and development of effective antifungal products.
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