Background: The growing problem of antimicrobial resistance motivated scientists to re-explore natural sources of bioactive secondary metabolites. Antarctica represents an extreme environment colonized by bacteria with unique adaptation mechanisms allowing them to thrive under harsh conditions. Such adaptations include production of secondary metabolites to inhibit competitors or sustain abiotic stresses, which predestines these microbes as source of natural products for biomedical use. Objectives: The aim of this work was to recover novel bacterial taxa from Antarctic soils to access the biosynthetic potential hidden in yet uncultivated bacteria. The main objective is the activation of silent biosynthetic gene clusters enabling discovery of novel secondary metabolites, mainly through co-cultivation strategies. Methods: Three isolation methods (pre-selection of spore-forming bacteria, low-nutrient and soil-extract based media) were applied to recover novel bacteria from Antarctic soils, predominantly targeting phyla with high biosynthetic potential such as Actinobacteriota, Proteobacteria and Firmicutes. Activation of silent biosynthetic gene clusters was attempted through targeted cultivation and co-cultivation. Screening for bioactive molecules and evaluation of their novelty was achieved by application of genomics, metabolomics, and bioactivity testing. Results: A collection of 917 isolates was established. Recovered isolates were associated with four bacterial phyla including 77 isolates of novel species. Proteobacteria and Actinobacteriota represented the most abundant phyla. Specific media stimulated biosynthesis of several unknown natural products. Eight strains produced antimicrobial compounds against resistant and multidrug-resistant bacterial and fungal pathogens. Importantly, metabolomic profiling indicated that these strains produced several new secondary metabolites, which may be responsible for the observed antimicrobial activities.