Antifungal activity of star anise extract against Penicillium roqueforti and Aspergillus niger for bread shelf life

Because star anise is underutilized in the baking sector and the antifungal targets are unclear, this study aimed to investigate the antifungal effect and mechanism of star anise extract (SAE) on spoilage fungi in bread. SAE was prepared by ethanol extraction and 31 substances were identified by GC-MS, among which trans-anethole (62.62%), estragole (7.82%) and linalool (4.66%) were the major components. The antifungal activity of SAE and the three main components against Penicillium roqueforti and Aspergillus niger were determined by using the Oxford cup method and the sesqui-dilution method. The inhibition zones were 9.88 mm and 15.09 mm, while the minimum inhibitory concentrations were 125.00 μL/mL and 31.25 μL/mL. Trans-anethole and estragole both showed antifungal activity against Penicillium roqueforti and Aspergillus niger, while linalool only showed antifungal activity against Aspergillus niger. Propidium iodide and fluorescein diacetate staining analysis, leakage of cellular components (nucleic acids and proteins) and rise in ergosterol content indicated that SAE disrupted the integrity and permeability of the cell membrane. Malondialdehyde was increased after SAE treatment, indicating that SAE caused lipid peroxidation in the cell membrane, further confirming that it disrupted the cell membrane. At the same time, SAE interacted with membrane proteins and altered their conformation, resulting in cell membrane dysfunction. Finally, the shelf life test showed that SAE extended the shelf life of the bread by up to 6 days. In general, this study highlights the antifungal effect of SAE against Penicillium roqueforti and Aspergillus niger, which indicated that SAE can be used as an antifungal agent to extend the shelf life of bread.