The Brazilian Caatinga Biome as a Hotspot for the Isolation of Antibiotic-Producing Actinomycetota

Antimicrobial resistance represents a critical global health challenge, intensifying the urgency of discovering novel antibiotics. <i>Actinomycetota</i> species, the most prolific source of clinical antibiotics, remain underexplored in unique ecosystems. In this study, we isolated 340 <i>Actinomycetota</i> strains from soils of the Brazilian semiarid Caatinga biome. Screening revealed that 122 isolates (35.9%) exhibited antimicrobial activity against clinically relevant pathogens (<i>Escherichia coli</i>, <i>Pseudomonas aeruginosa</i>, <i>Staphylococcus aureus</i>, and <i>Candida albicans</i>). Notably, 19 isolates showed activity against <i>Mycobacterium tuberculosis</i> H37Ra. MALDI-TOF MS analysis successfully provided genus-level identification for a subset of isolates, with approximately 32% assigned to the <i>Streptomyces</i> genus. However, the limited resolution of the database for the majority of the strains indicates high phylogenetic diversity and suggests the presence of potentially novel species. Metabolomic profiling via LC-MS/MS and GNPS molecular networking suggested the production of known antibiotics such as actinomycins, cyclomarins and anthracyclines and unveiled distinct molecular families putatively assigned to undescribed metabolites. Our work establishes the Caatinga biome as a valuable reservoir of bioactive <i>Actinomycetota</i>, encoding both known and potentially novel antimicrobial compounds. These results underscore the potential of underexplored and extreme environments in the quest to overcome antibiotic resistance.