Discovery of Novel Phenolic Compounds from Through OSMAC Approach: Structural Elucidation and Antibiotic Potential.

Among grapevine trunk diseases, Eutypa dieback, caused by the fungus , is one of the most critical ones, due to its widespread infection in vineyards and the lack of effective treatments. This fungus is a vascular pathogen that enters grapevines through pruning wounds. The infection process is associated with phytotoxic metabolites produced by the fungus, and as such, the identification of new metabolites from different culture conditions and broths could provide valuable insights into the fungus's enzymatic system and help its control. For the purposes of this study, the OSMAC (one strain, many compounds) approach was applied to investigate the secondary metabolism of strain 311 isolated from plants in Spain. A total of twenty metabolites were isolated, including five reported for the first time from and four that are newly identified compounds in the literature: eulatagalactoside A, ()-2-(4'-hydroxy-3'-methylbut-1'-yn-1'-yl)-4-(hydroxymethyl)phenol, ()-7-hydroxymethyl-3-methyl-2,3-dihydro-1-benzoxepin-3-ol, and (3a,4,5,7a)-4,5-dihydroxy-6-(()-3'-methylbuta-1',3'-dien-1'-ylidene)hexahydrobenzo[][1,3]dioxol-2-one. These compounds were extracted from fermentation broths using silica gel column chromatography and high-performance liquid chromatography (HPLC). Their structures were elucidated through extensive 1D and 2D NMR spectroscopy, along with high-resolution electrospray ionization mass spectrometry (HRESIMS). Compounds were evaluated for phytotoxicity against , with only eulatagalactoside A producing white spots after 48 h. Additionally, the antibacterial activity against , , and of selected compounds was tested. The compounds ()-2-(4'-hydroxy-3'-methylbut-1'-yn-1'-yl)-4-(hydroxymethyl)phenol and ()-7-(hydroxymethyl)-3-methyl-2,3-dihydrobenzo[]oxepin-3-ol showed the most significant antimicrobial activity against Gram-positive bacteria, inhibiting by over 75%, with IC values of 511.4 µg/mL and 617.9 µg/mL, respectively.