Background: The high salt concentration is the major factor limiting microbial growth at salterns, along with solar radiation, temperature, and pH. These environmental factors play key roles in the acquisition of unique genetic adaptations for the survival of microorganisms in salterns, which can result in the production of interesting secondary metabolites. The main goal of the present work was to isolate and compare the culturable microbiota from two geographically distant salterns in Portugal and access their biotechnological potential. Methods: Culturomics approaches using different culture media were applied for microbial isolation. All isolates were identified either by 16S rRNA or ITS genes sequencing, and their biotechonological potential was assessed by PCR. Results: Overall, 154 microbial isolates were recovered that were phylogenetically assigned to 45 taxa from 9 different phyla. From these, 26 isolates may represent putative new taxa. The predominant genera obtained were Penicillium (41 isolates, 26.6%), Streptomyces (13 isolates, 8.4%) and Sinomicrobium (11 isolates, 7.1%). Moreover, the polyketide synthase I gene was present in 64 isolates, the nonribosomal peptide synthethase gene in 16 isolates, and both genes in 23 isolates. Conclusions: This study adds up valuable knowledge on the culturable microbiota of Portuguese salterns and on its potential for production of secondary metabolites. In the long run, this study provides a widely diverse microbial collection for future works. Data public repository: All DNA sequences were deposited in the GenBank database at National Centre for Biotechnology Information (NCBI) web platform under accession numbers OK169439-OK169485, OK216020-OK216124, OK287059 and OK326927.