Targeted HRMS²-GNPS-based metabolomic analysis ofPseudoxylariasp. X187, a fungal antagonist of the fungus-growing termite symbiosis, resulted in the identification of two lipopeptidic congeners of xylacremolides, named xylacremolide C and D, which are built fromd-phenylalanine,l-proline and an acetyl-CoA starter unit elongated by four malonyl-CoA derived ketide units. The putativexyagene cluster was identified from a draft genome generated by Illumina and PacBio sequencing and RNAseq studies. Biological activities of xylacremolide A and B were evaluated and revealed weak histone deacetylase inhibitory (HDACi) and antifungal activities, as well as moderate protease inhibition activity across a panel of nine human, viral and bacterial proteases.