Halophilic and Halotolerant Bacteria from Dead Sea–Adjacent Soils as Antibiotic Producers: A Jordan-Centered Bioprospecting and Genome–Metabolome Discovery Framework

Abstract

Antimicrobial resistance continues to outpace the discovery of new antibiotics, motivating renewed exploration of under?sampled ecological niches. The Dead Sea region in Jordan is a polyextreme system characterized by high salinity (>34%), an unusual ionic composition enriched in divalent cations, and intense ultraviolet exposure—conditions that select for halophilic and halotolerant microorganisms with distinctive stress?adaptation biology and metabolite repertoires. Here we formulate a Jordan?centered discovery framework that integrates (i) stratified soil sampling from Dead Sea–adjacent gradients (salinity, moisture, vegetation, and anthropogenic influence), (ii) cultivation using high?salt selective media to enrich halophiles and halotolerant taxa, (iii) phenotypic antimicrobial screening against a clinically relevant pathogen panel, (iv) One Strain–Many Compounds (OSMAC) perturbations to activate silent biosynthetic pathways, and (v) paired genome mining and LC?MS/MS dereplication to prioritize novelty. Recent reviews emphasize hypersaline habitats as promising reservoirs of antibiotic chemistry, but also highlight rediscovery risks and the need for modern prioritization pipelines. Accordingly, our workflow couples antiSMASH?based biosynthetic gene cluster (BGC) analysis with molecular networking and dereplication tools to rapidly exclude known scaffolds and focus resources on new chemical space. We propose reporting standards (site metadata, growth conditions, activity metrics, and sequence?verified strain deposition) and a translational roadmap from crude extracts to purified leads, mechanism?of?action hypothesis generation, and early toxicity flags. This article is designed as a practical protocol?style blueprint to accelerate antibiotic discovery from Dead Sea–adjacent soils and to position Jordan as a regional hub for extremophile bio-discovery.