Mulberrin inhibits for strawberry storage by interfering with the bioactivity of 14α-demethylase (CYP51).

Currently, chemical agents hold great promise in preventing and combating . However, the antifungal mechanism of some agents for remains rather vague, imposing restrictions on the research and development of novel antifungal inhibitors. In this work, we discovered that mulberrin (MBN), a natural compound from the root bark of , with an IC of 1.38 μM together, demonstrated marked anti-14α-demethylase (CYP51) activity through high throughput virtual screening and bioactivity assay. The computational biology results demonstrated that MBN and its derivatives were bound to the catalytic activity region of CYP51, but only MBN could form a strong π-cation interaction with the Fe ion of heme in CYP51 the 2-methylpent-2-ene moiety at atom C9. MBN had a stronger binding free energy than the other three compounds with CYP51, implying that the 2-methylpent-2-ene moiety at atom C9 is a critical pharmacophore for CYP51 inhibitors. Subsequently, through an antifungal test, MBN demonstrated excellent anti- activity by inhibiting CYP51 activity. The EC values of MBN toward hyphal growth and spore germination in were 17.27 and 9.56 μg mL, respectively. The bioactivity loss of CYP51 by direct interaction with MBN induced the increase of cell membrane permeability, membrane destruction, and cell death. Meanwhile, in the infection model, MBN significantly prolonged the preservation of strawberries by preventing from infecting strawberries and could be used as a potential natural preserving agent for storing fruits.