Abstract
SummaryThe advent of metagenomics has greatly facilitated the discovery of enzymes with useful biochemical characteristics for industrial and biomedical applications, from environmental niches. In this study, we used sequence‐based metagenomics to identify two antibiotic resistance enzymes from the secluded, lower convective layer of Atlantis II Deep Red Sea brine pool (68°C, ~2200 m depth and 250‰ salinity). We assembled > 4 000 000 metagenomic reads, producing 43 555 contigs. Open reading frames (ORFs) called from these contigs were aligned to polypeptides from the Comprehensive Antibiotic Resistance Database using BLASTX. Two ORFs were selected for further analysis. The ORFs putatively coded for 3′‐aminoglycoside phosphotransferase [APH(3′)] and a class A beta‐lactamase (ABL). Both genes were cloned, expressed and characterized for activity and thermal stability. Both enzymes were active in vitro, while only APH(3′) was active in vivo. Interestingly, APH(3′) proved to be thermostable (T m = 61.7°C and ~40% residual activity after 30 min of incubation at 65°C). On the other hand, ABL was not as thermostable, with a T m = 43.3°C. In conclusion, we have discovered two novel AR enzymes with potential application as thermophilic selection markers.
Highlights
Red Sea brine pools represent a unique extreme and secluded environment to understand the evolution of biological life (Miller et al, 1966)
Salinity and temperature gradients segregate the brine into four layers, the lower convective layer (LCL) and three upper convective layers
Antimicrobial resistance genes have been previously identified in marine aquatic environments with no documented anthropogenic impact (Wegley et al, 2007; Toth et al, 2010)
Summary
Red Sea brine pools represent a unique extreme and secluded environment to understand the evolution of biological life (Miller et al, 1966). Atlantis II Deep (ATIID) is the largest and the most intriguing pool because of the multitude of extreme conditions. It has an area of 60 km and a salinity that is more than seven times that of normal sea water. The extreme conditions in LCL stimulated the search for extremophilic organisms and enzymes in this unique environment (Mohamed et al, 2013; Sayed et al, 2014; Sonbol et al, 2016). The properties of these enzymes could explain how indigenous microorganisms have evolved to survive such harsh environmental conditions and could reveal tools for several biotechnological applications
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