Untargeted metabolomics profiling of Morus alba twigs extract and antibacterial activity of pure metabolites

Plant natural products offer promise in combating multi-drug resistance by acting as antibacterial agents through various mechanisms. A metabolomic-guided phytochemical investigation, based on a LC-HRMS/MS and Molecular Networking combined approach, was carried out on an extract of M. alba L. (mulberry) twigs, a common of byproduct of mulberry utilization. Molecular Networking uncovered different clusters of prenylated polyphenols, glycosylated phenolic compounds, and Diels-Alder dimers, steering the phytochemical profiling of this extract. This led to the swift annotation and subsequent isolation of 17 secondary metabolites including stilbenoids, flavonoids, and flavanones. Isolated metabolites were tested for their antimicrobial activity against Staphylococcus species. The most active compound resulted to be kuwanon C, exhibiting a MIC value of 8 μg/mL against the methicillin resistant S. aureus (MRSA) strain and a biofilm producer strain of S. epidermidis. We also observed an interaction between 4 μg/mL of kuwanon C in combination with low oxacillin dosage against the MRSA. Thanks to the high chemical structure similarity of isolated metabolites, structure-activity relationships of these versatile scaffolds have been postulated.