Search

UNLABELLED: Biodegradable plastics are urgently needed to replace petroleum-derived polymeric materials and prevent their accumulation in the environment. To this end, we isolated and characterized a halophilic and alkaliphilic bacterium from the Great Salt Lake in Utah. The isolate was identified as a Halomonas species and designated CUBES01. Full-genome sequencing and genomic reconstruction revealed the unique genetic traits and metabolic capabilities of the strain, including the common polyhydroxyalkanoate (PHA) biosynthesis pathway. Fluorescence staining identified intracellular polyester granules that accumulated predominantly during the strains exponential growth, a feature rarely found among natural PHA producers. CUBES01 was found to metabolize a range of renewable carbon feedstocks, including glucosamine and acetyl-glucosamine, as well as sucrose, glucose, fructose, and further glycerol, propionate, and acetate. Depending on the substrate, the strain accumulated up to ~60% of its biomass (dry wt/wt) in poly(3-hydroxybutyrate), while reaching a doubling time of 1.7 h at 30°C and an optimum osmolarity of 1 M sodium chloride and a pH of 8.8. The physiological preferences of the strain may not only enable long-term aseptic cultivation but also facilitate the release of intracellular products through osmolysis. The development of a minimal medium also allowed the estimation of maximum polyhydroxybutyrate production rates, which were projected to exceed 5 g/h. Finally, also, the genetic tractability of the strain was assessed in conjugation experiments: two orthogonal plasmid vectors were stable in the heterologous host, thereby opening the possibility of genetic engineering through the introduction of foreign genes. IMPORTANCE: The urgent need for renewable replacements for synthetic materials may be addressed through microbial biotechnology. To simplify the large-scale implementation of such bio-processes, robust cell factories that can utilize sustainable and widely available feedstocks are pivotal. To this end, non-axenic growth-associated production could reduce operational costs and enhance biomass productivity, thereby improving commercial competitiveness. Another major cost factor is downstream processing, especially in the case of intracellular products, such as bio-polyesters. Simplified cell-lysis strategies could also further improve economic viability.

Terrestrial mud volcanoes represent surface features of channels for subsurface methane transport and, therefore, constitute an important source of methane emission from natural environments. How microbial processes regulate methane emissions in terrestrial mud volcanoes has yet to be fully addressed. This study demonstrated the geochemical characteristics and microbial communities of four mud volcano and seep sites in two geological settings of Sicily, Italy. At sites within the accretionary wedge that exhibited higher methane and sulfate concentrations, the communities were dominated by members capable of catalyzing methane and sulfate metabolisms and organic degradation. In particular, both anaerobic and aerobic methanotrophs were abundant and their abundance distribution coincided with the geochemical transition. In contrast, the sites near Mount Etna were characterized by high fluid salinity, CO2, and low methane and sulfate concentrations, with communities consisting of halophilic organic degraders and sulfur metabolizers, along with a minor presence of aerobic methanotrophs. Substantial variations in community composition and geochemistry across spatial and vertical redox gradients suggest that physicochemical contexts imposed by the geology, fluid path, and source characteristics play a vital role in shaping community composition and cycling of methane, sulfur and organic carbon in Sicily mud volcanoes. [ABSTRACT FROM AUTHOR]

Background: Polyhydroxyalkanoates (PHAs) are optimal potential materials for industrial and medical uses, characterized by exceptional sustainability, biodegradability, and biocompatibility. These are primarily from various bacteria and archaea. Bacterial strains with effective PHA formation capabilities and minimal production cost form the foundation for PHA production. Detailed genomic analysis of these PHA-generating bacteria is vital to understand their PHA production pathways and enhance their synthesis capability. Results: ZZQ-149, a halophilic, PHA-producing bacterium, was isolated from the sediment of China's Qinghai Lake. Here, we decoded the full genome of ZZQ-149 using Single Molecule Real Time (SMRT) technology based on PacBio RS II platform, coupled with Illumina sequencing platforms. Physiological, chemotaxonomic traits, and phylogenetic analysis based on 16 S rRNA gene and single copy core genes of ninety-nine Halomonas type strains identified ZZQ-149 as the type strain of Halomonas qinghailakensis. Furthermore, a low average nucleotide identity (ANI, < 95%) delineated the genetic differences between ZZQ-149 and other Halomonas species. The ZZQ-149 genome, with a DNA G + C content of 52%, comprises a chromosome (3, 798, 069 bps) and a plasmid (6, 107 bps). The latter encodes the toxin-antitoxin system, BrnT/BrnA. Through comprehensive genome sequencing and analysis, we identified multiple PHA-synthesizing enzymes and an unprecedented combination of eight PHA-synthesizing pathways in ZZQ-149. Conclusions: Being a halophilic, PHA-producing bacterium, ZZQ-149 exhibits potential as a high PHA producer for engineered bacteria via genome editing while ensuring low-cost PHA production through continuous, unsterilized fermentation. [ABSTRACT FROM AUTHOR]

Abstract Background Polyhydroxyalkanoates (PHAs) are optimal potential materials for industrial and medical uses, characterized by exceptional sustainability, biodegradability, and biocompatibility. These are primarily from various bacteria and archaea. Bacterial strains with effective PHA formation capabilities and minimal production cost form the foundation for PHA production. Detailed genomic analysis of these PHA-generating bacteria is vital to understand their PHA production pathways and enhance their synthesis capability. Results ZZQ-149, a halophilic, PHA-producing bacterium, was isolated from the sediment of China’s Qinghai Lake. Here, we decoded the full genome of ZZQ-149 using Single Molecule Real Time (SMRT) technology based on PacBio RS II platform, coupled with Illumina sequencing platforms. Physiological, chemotaxonomic traits, and phylogenetic analysis based on 16 S rRNA gene and single copy core genes of ninety-nine Halomonas type strains identified ZZQ-149 as the type strain of Halomonas qinghailakensis. Furthermore, a low average nucleotide identity (ANI,

Biodegradable plastics are urgently needed to replace petroleum-derived polymeric materials and prevent their accumulation in the environment. To this end, we isolated and characterized a halophilic and alkaliphilic bacterium from the Great Salt Lake in Utah. The isolate was identified as a Halomonas species and designated "CUBES01." Full-genome sequencing and genomic reconstruction revealed the unique genetic traits and metabolic capabilities of the strain, including the common polyhydroxyalkanoate (PHA) biosynthesis pathway. Fluorescence staining identified intracellular polyester granules that accumulated predominantly during the strain's exponential growth, a feature rarely found among natural PHA producers. CUBES01 was found to metabolize a range of renewable carbon feedstocks, including glucosamine and acetyl-glucosamine, as well as sucrose, glucose, fructose, and further glycerol, propionate, and acetate. Depending on the substrate, the strain accumulated up to ~60% of its biomass (dry wt/wt) in poly(3-hydroxybutyrate), while reaching a doubling time of 1.7 h at 30°C and an optimum osmolarity of 1 M sodium chloride and a pH of 8.8. The physiological preferences of the strain may not only enable long-term aseptic cultivation but also facilitate the release of intracellular products through osmolysis. The development of a minimal medium also allowed the estimation of maximum polyhydroxybutyrate production rates, which were projected to exceed 5 g/h. Finally, also, the genetic tractability of the strain was assessed in conjugation experiments: two orthogonal plasmid vectors were stable in the heterologous host, thereby opening the possibility of genetic engineering through the introduction of foreign genes. [ABSTRACT FROM AUTHOR]

The extremophile Halobacterium salinarum is an aerobic archaeon that has adapted to thrive in high-salt environments such as salted fish, hypersaline lakes, and salterns. Halophiles have garnered significant interest due to their unique interactions with bacteriophages known as haloarchaeophages. Studies have identified and characterized prophages in halophilic archaea, such as Haloferax volcanii, Haloquadratum walsbyi, and Haloarcula marismortui. Still, an investigation has yet to be conducted into the presence of prophage elements on Halobacterium salinarum ATCC 33170. This is of particular interest to us as we are using this strain as a source of archaeol, as one of the components of our sulfated lactosyl archaeol (SLA) archaeosome adjuvant. Genomic contigs of strain 33170 were bioinformatically assessed for prophage-like features using BLAST, PHASTER, InterProScan, and PHYRE2. A 7 kb region encoding six genes was identified as an incomplete prophage, and the proteins were further analyzed, revealing high homology to proteins encoded by bacteria, archaea, and an IS200 transposon. Restricting the BLASTp database to viruses resulted in hits to both myo- and siphoviral proteins, which would be unusual for an intact prophage. Additionally, no known phage structural proteins were identified in the search, suggesting a low chance that H. salinarum ATCC 33170 harbors a latent prophage. [ABSTRACT FROM AUTHOR]

CetZ proteins are archaea-specific homologs of the cytoskeletal proteins FtsZ and tubulin. In the pleomorphic archaeon Haloferax volcanii, CetZ1 contributes to the development of rod shape and motility, and has been implicated in the proper assembly and positioning of the archaellum and chemotaxis motility proteins. CetZ1 shows complex subcellular localization, including irregular midcell structures and filaments along the long axis of developing rods and patches at the cell poles of the motile rod cell type. The polar localizations of archaellum and chemotaxis proteins are also influenced by MinD4, the only previously characterized archaeal member of the MinD family of ATPases, which are better known for their roles in the positioning of the division ring in bacteria. Using minD mutant strains and CetZ1 subcellular localization studies, we show here that a second minD homolog, minD2, has a strong influence on motility and the localization of CetZ1. Knockout of the minD2 gene altered the distribution of a fluorescent CetZ1-mTq2 fusion protein in a broad midcell zone and along the edges of rod cells, and inhibited the localization of CetZ1-mTq2 at the cell poles. MinD4 had a similar but weaker influence on motility and CetZ1-mTq2 localization. The minD2/4 mutant strains formed rod cell shapes like the wildtype at an early log stage of growth. Our results are consistent with distinct roles for CetZ1 in rod shape formation and at the poles of mature rods, that are positioned through the action of the MinD proteins and contribute to the development of swimming motility in multiple ways. They represent the first report of MinD proteins controlling the positioning of tubulin superfamily proteins in archaea.

The extremophile Halobacterium salinarum is an aerobic archaeon that has adapted to thrive in high-salt environments such as salted fish, hypersaline lakes, and salterns. Halophiles have garnered significant interest due to their unique interactions with bacteriophages known as haloarchaeophages. Studies have identified and characterized prophages in halophilic archaea, such as Haloferax volcanii, Haloquadratum walsbyi, and Haloarcula marismortui. Still, an investigation has yet to be conducted into the presence of prophage elements on Halobacterium salinarum ATCC 33170. This is of particular interest to us as we are using this strain as a source of archaeol, as one of the components of our sulfated lactosyl archaeol (SLA) archaeosome adjuvant. Genomic contigs of strain 33170 were bioinformatically assessed for prophage-like features using BLAST, PHASTER, InterProScan, and PHYRE2. A 7 kb region encoding six genes was identified as an incomplete prophage, and the proteins were further analyzed, revealing high homology to proteins encoded by bacteria, archaea, and an IS200 transposon. Restricting the BLASTp database to viruses resulted in hits to both myo- and siphoviral proteins, which would be unusual for an intact prophage. Additionally, no known phage structural proteins were identified in the search, suggesting a low chance that H. salinarum ATCC 33170 harbors a latent prophage.

The bacterium Natranaerobius thermophilus is an extremely halophilic alkalithermophile that can thrive under conditions of high salinity (3.3-3.9 M Na+), alkaline pH (9.5), and elevated temperature (53°C). To understand the molecular mechanisms of salt adaptation in N. thermophilus, it is essential to investigate the protein, mRNA, and key metabolite levels on a molecular basis. Based on proteome pro filing of N. thermophilus under 3.1, 3.7, and 4.3 M Na+ conditions compared to 2.5 M Na+ condition, we discovered that a hybrid strategy, combining the "compatible solute" and "salt-in" mechanisms, was utilized for osmotic adjustment dur ing the long-term salinity adaptation of N. thermophilus. The mRNA level of key proteins and the intracellular content of compatible solutes and K+ support this conclusion. Specifically, N. thermophilus employs the glycine betaine ABC transporters (Opu and ProU families), Na+/solute symporters (SSS family), and glutamate and proline synthesis pathways to adapt to high salinity. The intracellular content of compatible solutes, including glycine betaine, glutamate, and proline, increases with rising salinity levels in N. thermophilus. Additionally, the upregulation of Na+/K+/H+ transporters facilitates the maintenance of intracellular K+ concentration, ensuring cellular ion homeostasis under varying salinities. Furthermore, N. thermophilus exhibits cytoplasmic acidification in response to high Na+ concentrations. The median isoelectric points of the upregulated proteins decrease with increasing salinity. Amino acid metabolism, carbohydrate and energy metabolism, membrane transport, and bacterial chemotaxis activities contribute to the adaptability of N. thermophilus under high salt stress. This study provides new data that support further elucidating the complex adaptation mechanisms of N. thermophilus under multiple extremes. [ABSTRACT FROM AUTHOR]

A single extremely halophilic strain was isolated from salt brine produced when a fresh water lake flooded a large salt mine located beneath the lake. The water that entered this mine contained less than 0.34 M NaCl, but over time, this sealed brine became saturated by Cenozoic age salt (121–125 million-year BCE). The isolated strain requires at least 1.7 M NaCl for survival and grows optimally in 3.1 M NaCl. Therefore, it could not have survived or been present in the waters that flooded this salt mine. The strain grows at a pH range from 6.5 to 9.0 and has a wide tolerance to temperatures from 25 ℃ to at least 60 ℃. The comparison of 16S rRNA and rpoB′ genes revealed that strain 1–13-28T is related to Halorubrum tebenquichense DSM 14210T showing 98.6% and 98.1% similarities, respectively. Phylogenetic analyses based on 16S rRNA, rpoB′ genes and 122 concatenated archaeal genes show that the strain 1–13-28T consistently forms a cluster with Halorubrum tebenquichense of the genus Halorubrum. Strain 1–13-28T contained sulfated mannosyl glucosyl diether, and the polar lipid profile was identical to those of most Halorubrum species. Based on the overall combination of physiological, phylogenetic, polar lipids and phylogenomic characteristics, strain 1–13-28T (= ATCC 700083T = CGMCC 1.62627T) represents a newly identified species within the genus Halorubrum for which the name Halorubrum hochsteinianum is proposed. [ABSTRACT FROM AUTHOR]

Microorganisms in hypersaline potash mining byproducts and their potential environmental applications have not been extensively reported. This study reports the diverse waste-impacted microbial communities (archaea and bacteria) adapted to extreme salinity (>10–25%). Of these, halotolerant Croceicoccus sp. FTI14 was investigated as a biosorbent for removing dissolved Cu(II) and Cr(VI) from synthetic Cu(II) and Cr(VI)-contaminated DI, groundwater and saline groundwater (0.55 M ionic strength). FTI14 biomass was oven-dried, finely ground, and investigated in batch biosorption experiments. At initial metal concentrations of 40 mg/L, FTI14 removed 40 ± 0.7% (16.3 ± 0.5 mg/g) and 19 ± 0.1% (7.8 ± 0.1 mg/g) of the Cu(II) from deionized water and saline groundwater, respectively, while only 22.9 ± 0.7% (9.6 ± 0.2 mg/g) and 2.1 ± 0.6% (1.0 ± 0.3 mg/g) Cr(VI) removal was achieved. Cu(II) uptake (mg/g) exceeded Cr(VI) uptake by a factor of 1.7–7.8. Langmuir and Freundlich models were applied on FTI14 biosorption isotherm data. The Freundlich model showed a better fit for both Cu(II) and Cr(VI), as indicated by the AIC values compared with evidence ratios. Synchrotron-based scanning transmission X-ray microscopy (STXM) visualizations of the biosorbent showed a mixture of whole cells and indistinct biomass and a spatial association between metals and biomass. Metal exposure alters the amide functional groups peak in Fourier transform Infra-red (FTIR) spectra, suggesting its role in sorption process. Thus, this study indicates culturable halotolerant microorganisms from hypersaline potash mining byproducts and its potential as biosorbent applications for metal removal from impacted groundwater.

A haloalkaliphilic actinobacterial strain LS1_29T, isolated from an inland hypersaline Sambhar Salt Lake, situated in Rajasthan, India, was subjected to taxonomic characterisation using the polyphasic approach. Cells of the strain were Gram stain positive and aerobic, having reticulate and aerial hyphae. The major fatty acids detected were iso C16:0 (38.23%), anteiso C17:0 (20.52%), iso C18:0 10 methyl (8.09%), iso C18:0 (7.74%) and iso C17:0 (7.48%). The major polar lipids identified were diphosphatidyl glycerol, phosphatidylglycerol, phosphatidylcholine and phospholipids. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the strain belonged to the genus Lipingzhangella with the highest similarity of 94.09% to Lipingzhangella halophila EGI 80537 T and formed a separate clade within the genus Lipingzhangella. The DNA G + C content of genomic DNA of strain LS1_29T was 67.99 mol%. The strain showed the highest orthologous average nucleotide identity (OrthoANI) value of 72.80% and digital DNA–DNA hybridization (dDDH) value of 20.3% with Lipingzhangella halophila EGI 80537 T. Based on the phenotypic, chemotaxonomic and phylogenetic characteristics, strain LS1_29T represents a novel species within the genus Lipingzhangella, for which the name Lipingzhangella rawalii sp. nov. is proposed. The type strain of the type species is LS1_29T (= KCTC 49199 T = JCM 32979 T = MCC 3420 T). [ABSTRACT FROM AUTHOR]

This study aimed to determine salinity tolerances in Coxiella gastropods from Australian salt lakes and whether different species exhibit characteristically different tolerances. Controlled gradual accumulation experiments were conducted to estimate both the maximum and minimum salinity levels at which 50% of individuals (IC50) remained active for 25 populations representing six species. All studied species showed remarkable euryhalinity and were tolerant of very high levels of salinity, some more than others, while minimum salinity tolerance varied little among populations and species. The experimental trends in salinity tolerances were consistent with the salinity distributions of species in the field, although the former were typically broader than latter. The findings suggest that Coxiella comprises some of the most salt tolerant gastropods globally. [ABSTRACT FROM AUTHOR]

Cyanobacteria that grow above seawater salinity at temperatures above 45°C have rarely been studied.The present study is an attempt to decipher these unknown facts where the unique properties of phosphatase enzymes in a thermo-halotolerant Iceland clone 2 Leptolyngbyahas been studied under some important environmental conditions that play a distinctive rolein the growth of these extremophiles in those adverse ecological niches. Leptolyngbyaused in this present study is a unique species having 2 extreme characteristics of tolerance to high salt concentrations and temperature, so it is of intrinsic and scientific interest to study the phosphate dynamics and its variability under different ecological factors.The results of these experiments clearly show that under very adverse conditions of low light or high temperature and very high salt concentrations (almost 3 times of salt present in seawater 90g/L) this extremophile has the capacity to maintain its growth and metabolism which is the key to its survival in these extreme habitats.Observations from growth experiments under different environmental conditions(Temperatures, pH, salt concentrations, different light intensities) under laboratory conditions were found to be like its diverse patterns and adaptive ability in the extreme environment this organism has been isolated from. Phosphatase activity as a wayof understanding how P is metabolized under extreme conditions revealed that the highest phosphatase activity was observed in high salt concentrations (3 times that of seawater) and high temperatures of 45°C and low light intensities that is a very significant observation and scientifically important.

While life on Mars has not been found, Earth-based microorganisms may contaminate the Red Planet during rover expeditions and human exploration. Due to the survival advantages conferred by the biofilm morphology to microorganisms, such as resistance to UV and osmotic stress, biofilms are particularly concerning from a planetary protection perspective. Modeling and data from the NASA Phoenix mission indicate that temporary liquid water might exist on Mars in the form of high salinity brines. These brines could provide colonization opportunities for terrestrial microorganisms brought by spacecraft or humans. To begin testing for potential establishment of microbes, results are presented from a simplified laboratory model of a Martian saline seep inoculated with sediment from Hailstone Basin, a terrestrial saline seep in Montana (USA). The seep was modeled as a sand-packed drip flow reactor at room temperature fed media with either 1 M MgSO4 or 1 M NaCl. Biofilms were established within the first sampling point of each experiment. Endpoint 16S rRNA gene community analysis showed significant selection of halophilic microorganisms by the media. Additionally, we detected 16S rRNA gene sequences highly similar to microorganisms previously detected in two spacecraft assembly cleanrooms. These experimental models provide an important foundation for identifying microbes that could hitch-hike on spacecraft and may be able to colonize Martian saline seeps. Future model optimization will be vital to informing cleanroom sterilization procedures.

Xinghamide A (1), a new nonapeptide, was discovered in Streptomyces xinghaiensis isolated from a halophyte, Suaeda maritima (L.) Dumort. Based on high-resolution mass and NMR spectroscopic data, the planar structure of 1 was established, and, in particular, the sequence of nine amino acids was determined with ROESY and HMBC NMR spectra. The absolute configurations of the α-carbon of each amino acid residue were determined with 1-fluoro-2,4-dinitrophenyl-l-and -d-leucine amide (Marfey's reagents) and 2,3,4,6-tetra-O-acetyl-β-d-glucopyranosyl isothiocyanate, followed by LC-MS analysis. The anti-inflammatory activity of xinghamide A (1) was evaluated by inhibitory abilities against the nitric oxide (NO) secretion and cyclooxygenase-2 (COX-2) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. [ABSTRACT FROM AUTHOR]

Hahella is a genus that has not been well-studied, with only two identified species. The potential of this genus to produce cellulases is yet to be fully explored. The present study isolated Hahella sp. CR1 from mangrove soil in Tanjung Piai National Park, Malaysia, and performed whole genome sequencing (WGS) using NovaSeq 6000. The final assembled genome consists of 62 contigs, 7,106,771 bp, a GC ratio of 53.5%, and encoded for 6,397 genes. The CR1 strain exhibited the highest similarity with Hahella sp. HN01 compared to other available genomes, where the ANI, dDDH, AAI, and POCP were 97.04%, 75.2%, 97.95%, and 91.0%, respectively. In addition, the CAZymes analysis identified 88 GTs, 54 GHs, 11 CEs, 7 AAs, 2 PLs, and 48 CBMs in the genome of strain CR1. Among these proteins, 11 are related to cellulose degradation. The cellulases produced from strain CR1 were characterized and demonstrated optimal activity at 60 ℃, pH 7.0, and 15% (w/v) sodium chloride. The enzyme was activated by K+, Fe2+, Mg2+, Co2+, and Tween 40. Furthermore, cellulases from strain CR1 improved the saccharification efficiency of a commercial cellulase blend on the tested agricultural wastes, including empty fruit bunch, coconut husk, and sugarcane bagasse. This study provides new insights into the cellulases produced by strain CR1 and their potential to be used in lignocellulosic biomass pre-treatment. [ABSTRACT FROM AUTHOR]

The present study was aimed to isolate a marine bacterium from biofilms with promising lignin peroxidase production potential. Biofilm samples were collected from the surfaces of boats, concrete structures and mangrove trees submerged during high tide and exposed during low tide in the Parangipettai area of Tamil Nadu, India. Collected biofilm samples were used for the isolation of bacteria and screened of lignin peroxidase (LiP) activity. Among the 106 bacterial isolates collected, the strain designated as PBB09 isolated from the boat biofilm showed highest LiP activity was identified as Salinococcus roseus. Optimization of the S. roseus strain for growth and LiP production revealed 36 hrs of incubation period, 100 rpm agitation, pH 8, 35°C, 5% salinity, 0.3% lignin, 2% glucose as the carbon source and 1% yeast extract as the nitrogen source were the ideal conditions. When three different cheaper substrates were tested (from 1-5%), corn cobs (at 2%) gave a maximum growth of 2.44 OD and 47 U/ml/min. of lignin peroxidase production, followed by sugarcane bagasse (at 3%) with 2.18 OD and 40 U/ml/min. and rice straw (at 4%) with 1.75 OD and 26 U/ml/min. Mass scale cultivation in shake flask with the above conditions gave a maximum growth of 4.16 OD with the lignin peroxidase activity of 93 U/ml/min. with the standard substrates (without lignin) whereas it was 3.2 OD with the lignin peroxidase activity of 63 U/ml/min. when using corn cobs as a cheaper substrate. In fermentor, the observed growth and lignin peroxidase activity were 6.7 OD and 119 U/ml/min. for the standard substrate while, it was 4.5 OD and 86 U/ml/min. with the cheaper substrate (i.e.) 2% corn cobs. [ABSTRACT FROM AUTHOR]

Biologically enhanced transport of radionuclides is one of several processes that can affect the performance of a nuclear waste repository. In this work, several microbial isolates from the Waste Isolation Pilot Plant (WIPP) were tested for their influence on the concentration of neodymium, as an analog for +3 actinides, in simple sodium chloride solutions and in anoxic WIPP brines. Batch sorption experiments were carried out over a period of 4–5 weeks. In many cases, the effect on neodymium in solution was immediate and extensive and assumed to be due to surface complexation. However, over time, the continued loss of Nd from the solution was more likely due to biologically induced precipitation and/or mineralization and possible entrapment in extracellular polymeric substances. The results showed no correlation between organism type and the extent of its influence on neodymium in solution. However, a correlation was observed between different test matrices (simple NaCl versus high-magnesium brine versus high-NaCl brine). Further experiments were conducted to test these matrix effects, and the results showed a significant effect of magnesium concentration on the ability of microorganisms to remove Nd from solution. Possible mechanisms include cation competition and the alteration of cell surface structures. This suggests that the aqueous chemistry of the WIPP environs could play a larger role in the final disposition of +3 actinides than the microbiology. [ABSTRACT FROM AUTHOR]

Barkol Lake, situated northeast of the Tianshan Mountains, Xinjiang, is a hypersaline lake with abundant sulfate and chloride minerals, which can be a potential analog for microbial saline paleolakes on Mars. The lake water, sediments, and surrounding soils of Barkol Lake were sampled for geochemical analysis and 16S rRNA gene sequencing to investigate the prokaryotic community structure, abundances, interactions, and ecological functions. Results show that (1) prokaryotic community structure differs significantly between biotopes (water, sediment, and soil), with the highest abundances of archaea occurring in water samples and highest prokaryotic diversities in soil samples; (2) archaeal communities are dominated by Halobacterota, Nanoarchaeota, Thermoplasmatota, and Crenarchaeota, while bacterial communities are mainly Proteobacteria, Bacteroidetes, Actinobacteria, Desulfobacterota, Chloroflexi, Gemmatimonadetes, Firmicutes, and Cyanobacteria; (3) the prokaryotic community network for soil is far more complicated and stable than those for water and sediment; (4) soil prokaryotic communities could be significantly affected by environmental factors such as salinity, pH, total sulfur, and Ca2+; (5) archaeal communities may play an important role in the nitrogen cycle, while bacterial communities may mainly participate in the sulfur cycle. This study extends the data set of prokaryotic communities for hypersaline environments, which will provide perspectives into identification of the counterparts and help to understand potential microbial interactions and biogeochemical cycles occurring on Mars. [ABSTRACT FROM AUTHOR]

Halophilic archaea are the dominant type of microorganisms in hypersaline environments. The diversity of halophilic archaea in Zehrez-Chergui (Saharian chott) was analyzed and compared by both analysis of a library of PCR amplified 16S rRNA genes and by cultivation approach. This work, represents the first of its type in Algeria. A total cell count was estimated at 3.8 × 103 CFU/g. The morphological, biochemical, and physiological characterizations of 45 distinct strains, suggests that all of them might be members of the class Halobacteria. Among stains, 23 were characterized phylogenetically and are related to 6 genera of halophilic archaea.The dominance of the genus Halopiger, has not been reported yet in other hypersaline environments. The 100 clones obtained by the molecular approach, were sequenced, and analyzed. The ribosomal library of 61 OTUs showed that the archaeal diversity included uncultured haloarcheon, Halomicrobium, Natronomonas, Halomicroarcula, Halapricum, Haloarcula, Halosimplex, Haloterrigena, Halolamina, Halorubellus, Halorussus and Halonotius. The results of rarefaction analysis indicated that the analysis of an increasing number of clones would have revealed additional diversity. Surprisingly, no halophilic archaea were not shared between the two approaches. Combining both types of methods was considered the best approach to acquire better information on the characteristics of soil halophilic archaea. [ABSTRACT FROM AUTHOR]

Ultraviolet (UV) radiation responses of extremophilic and archaeal microorganisms are of interest from evolutionary, physiological, and astrobiological perspectives. Previous studies determined that the halophilic archaeon, Halobacterium sp. NRC-1, which survives in multiple extremes, is highly tolerant of UV radiation. Here, Halobacterium sp. NRC-1 UV tolerance was compared to taxonomically diverse Haloarchaea isolated from high-elevation salt flats, surface warm and cold hypersaline lakes, and subsurface Permian halite deposits. Haloterrigena/Natrinema spp. from subsurface halite deposits were the least tolerant after exposure to photoreactivating light. This finding was attributed to deviation of amino acid residues in key positions in the DNA photolyase enzyme or to the complete absence of the photolyase gene. Several Halobacterium, Halorubrum and Salarchaeum species from surface environments exposed to high solar irradiance were found to be the most UV tolerant, and Halorubrum lacusprofundi from lake sediment was of intermediate character. These results indicate that high UV tolerance is not a uniform character trait of Haloarchaea and is likely reflective of UV exposure experienced in their environment. This is the first report correlating natural UV tolerance to photolyase gene functionality among Haloarchaea and provides insights into their survival in ancient halite deposits and potentially on the surface of Mars. [ABSTRACT FROM AUTHOR]

Aims To develop a model HACCP plan related to the microbiological hazards for the traditional fermented crab. Methods and Results The microbiological and chemical characteristics of commercial products were surveyed. Microbiological hazard analysis was performed for raw materials and during processing. Critical control points (CCPs) were determined using a decision tree, with CCP1 as saturated salt preparation and CCP2 as fermentation. The critical limit (CL) of CCP1 was at 100°C for 20 min applied to brining and of CCP2 was at 25% NaCl for the brine applied to fermented crab. Isolated microbial hazards and type strains were used for the validation of the CLs. Monitoring and verification of the proposed HACCP plan were carried out, and an effective HACCP plan was established. Conclusions The HACCP plan promoted the safe consumption of fermented crab with the provided CCPs at the saturated salt preparation and fermentation steps. The effective CLs to ensure microbiological hazards as safe at the CCPs provide the best support for an effective plan. The hazards were reduced significantly after the HACCP plan had been applied. [ABSTRACT FROM AUTHOR]

The halophilic genus Joostella is one of the least-studied genera in the family of Flavobacteriaceae. So far, only two species were taxonomically identified with limited genomic analysis in the aspect of application has been reported. Joostella atrarenae M1-2T was previously isolated from a seashore sample and it is the second discovered species of the genus Joostella. In this project, the genome of J. atrarenae M1-2T was sequenced using NovaSeq 6000. The final assembled genome is comprised of 71 contigs, a total of 3,983,942 bp, a GC ratio of 33.2%, and encoded for 3,416 genes. The 16S rRNA gene sequence of J. atrarenae M1-2T shows 97.3% similarity against J. marina DSM 19592T. Genome-genome comparison between the two strains by ANI, dDDH, AAI, and POCP shows values of 80.8%, 23.3%, 83.4%, and 74.1% respectively. Pan-genome analysis shows that strain M1-2T and J. marina DSM 19592T shared a total of 248 core genes. Taken together, strain M-2T and J. marina DSM 19592T belong to the same genus but are two different species. CAZymes analysis revealed that strain M1-2T harbors 109 GHs, 40 GTs, 5 PLs, 9 CEs, and 6 AAs. Among these CAZymes, while 5 genes are related to cellulose degradation, 12 and 24 genes are found to encode for xylanolytic enzymes and other hemicellulases that involve majorly in the side chain removal of the lignocellulose structure, respectively. Furthermore, both the intracellular and extracellular crude extracts of strain M1-2T exhibited enzymatic activities against CMC, xylan, pNPG, and pNPX substrates, which corresponding to endoglucanase, xylanase, β-glucosidase, and β-xylosidase, respectively. Collectively, description of genome coupled with the enzyme assay results demonstrated that J. atrarenae M1-2T has a role in lignocellulosic biomass degradation, and the strain could be useful for lignocellulosic biorefining. [ABSTRACT FROM AUTHOR]

Cyanobacteria that grow above seawater salinity at temperatures above 45°C have rarely been studied. The present study is an attempt to decipher these unknown facts where the unique properties of phosphatase enzymes in a thermo-halotolerant Iceland clone 2 Leptolyngbya has been studied under some important environmental conditions that play a distinctive role in the growth of these extremophiles in those adverse ecological niches. Leptolyngbya used in this present study is a unique species having 2 extreme characteristics of tolerance to high salt concentrations and temperature, so it is of intrinsic and scientific interest to study the phosphate dynamics and its variability under different ecological factors. The results of these experiments clearly show that under very adverse conditions of low light or high temperature and very high salt concentrations (almost 3 times of salt present in seawater 90g/L) this extremophile has the capacity to maintain its growth and metabolism which is the key to its survival in these extreme habitats. Observations from growth experiments under different environmental conditions (Temperatures, pH, salt concentrations, different light intensities) under laboratory conditions were found to be like its diverse patterns and adaptive ability in the extreme environment this organism has been isolated from. Phosphatase activity as a way of understanding how P is metabolized under extreme conditions revealed that the highest phosphatase activity was observed in high salt concentrations (3 times that of seawater) and high temperatures of 45°C and low light intensities that is a very significant observation and scientifically important. [ABSTRACT FROM AUTHOR]

The polyextremophilic β-galactosidase enzyme of the haloarchaeon Halorubrum lacusprofundi functions in extremely cold and hypersaline conditions. To better understand the basis of polyextremophilic activity, the enzyme was studied using steady-state kinetics and molecular dynamics at temperatures ranging from 10 °C to 50 °C and salt concentrations from 1 M to 4 M KCl. Kinetic analysis showed that while catalytic efficiency (kcat/Km) improves with increasing temperature and salinity, Km is reduced with decreasing temperatures and increasing salinity, consistent with improved substrate binding at low temperatures. In contrast, kcat was similar from 2–4 M KCl across the temperature range, with the calculated enthalpic and entropic components indicating a threshold of 2 M KCl to lower the activation barrier for catalysis. With molecular dynamics simulations, the increase in per-residue root-mean-square fluctuation (RMSF) was observed with higher temperature and salinity, with trends like those seen with the catalytic efficiency, consistent with the enzyme's function being related to its flexibility. Domain A had the smallest change in flexibility across the conditions tested, suggesting the adaptation to extreme conditions occurs via regions distant to the active site and surface accessible residues. Increased flexibility was most apparent in the distal active sites, indicating their importance in conferring salinity and temperature-dependent effects. [ABSTRACT FROM AUTHOR]

This study was performed to find halophilic bacteria that have the ability to increase salinity resistance in wheat seedlings, so that any type of wheat cultivar can be grown in saline areas by inoculation with these bacteria without the need for genetic manipulation.A number of highly saline farm soils from several areas around Tabriz and Urmia Lake were screened to isolate bacterial isolates. A total of six highly halophilic isolates were isolated for the study. The effect of bacteria on salinity tolerance in wheat was determined as a factorial of two factors (salinity and bacteria) based on a randomized complete block design with three replications. The effect of six bacterial isolates at three high concentrations of salt (NaCl) (0.5, 1 and 1.5%) on wheat seed growth under standard laboratory conditions for four weeks was evaluated. Two bacterial isolates that had the best performance - in most growth indices such as fresh and dry size and weight of roots and stems - were selected and their effect was assayed under greenhouse conditions. Finally, the best bacteria in terms of function were selected and identified by molecular methodsFrom the six highly halophilic bacterial isolates, two bacteria were selected for greenhouse studies, one of which, caused a 50% increase in most growth factors, was selected for identification. Molecular identification based on barcoding 16S rDNA region showed that this bacterium belongs to Oceanbacillus picturae with 99% probability.This is the first report of the presence of Oceanbacillus picturae in the soils of

Xinghamide A (1), a new nonapeptide, was discovered in Streptomyces xinghaiensis isolated from a halophyte, Suaeda maritima (L.) Dumort. Based on high-resolution mass and NMR spectroscopic data, the planar structure of 1 was established, and, in particular, the sequence of nine amino acids was determined with ROESY and HMBC NMR spectra. The absolute configurations of the α-carbon of each amino acid residue were determined with 1-fluoro-2,4-dinitrophenyl-l-and -d-leucine amide (Marfey’s reagents) and 2,3,4,6-tetra-O-acetyl-β-d-glucopyranosyl isothiocyanate, followed by LC-MS analysis. The anti-inflammatory activity of xinghamide A (1) was evaluated by inhibitory abilities against the nitric oxide (NO) secretion and cyclooxygenase-2 (COX-2) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells.

Halophilic microorganisms have long been known to survive within the brine inclusions of salt crystals, as evidenced by the change in color for salt crystals containing pigmented halophiles. However, the molecular mechanisms allowing this survival has remained an open question for decades. While protocols for the surface sterilization of halite (NaCl) have enabled isolation of cells and DNA from within halite brine inclusions, “-omics” based approaches have faced two main technical challenges: (1) removal of all contaminating organic biomolecules (including proteins) from halite surfaces, and (2) performing selective biomolecule extractions directly from cells contained within halite brine inclusions with sufficient speed to avoid modifications in gene expression during extraction. In this study, we tested different methods to resolve these two technical challenges. Following this method development, we then applied the optimized methods to perform the first examination of the early acclimation of a model haloarchaeon (Halobacterium salinarum NRC-1) to halite brine inclusions. Examinations of the proteome of Halobacterium cells two months post-evaporation revealed a high degree of similarity with stationary phase liquid cultures, but with a sharp down-regulation of ribosomal proteins. While proteins for central metabolism were part of the shared proteome between liquid cultures and halite brine inclusions, proteins involved in cell mobility (archaellum, gas vesicles) were either absent or less abundant in halite samples. Proteins unique to cells within brine inclusions included transporters, suggesting modified interactions between cells and the surrounding brine inclusion microenvironment. The methods and hypotheses presented here enable future studies of the survival of halophiles in both culture model and natural halite systems.

Halomonas spp. are the well-studied platform organisms or chassis for next-generation industrial biotechnology (NGIB) due to their contamination-resistant nature combined with their fast growth property. Several Halomonas spp. have been studied regarding their genomic information and molecular engineering approaches. Halomonas spp., especially Halomonas bluephagenesis, have been engineered to produce various biopolyesters such as polyhydroxyalkanoates (PHA), proteins including surfactants and enzymes, small molecular compounds including amino acids and derivates, as well as organic acids. This paper reviews all the progress reported in the last 10 years regarding this robust microbial cell factory. Key points: • Halomonas spp. are robust chassis for low-cost production of chemicals • Genomic information of some Halomonas spp. has been revealed • Molecular tools and approaches for Halomonas spp. have been developed • Halomonas spp. are becoming more and more important for biotechnology [ABSTRACT FROM AUTHOR]

Halomonas smyrnensis AAD6T is a moderately halophilic bacterium proven to be a powerful biotechnological tool with its ability to accumulate valuable biopolymers such as levan and poly(3-hydroxybutyrate) (PHB). Levan is a fructose homopolymer with β-2,6 fructofuranosidic linkages on the polymer backbone, and its distinctive applications in various industries such as food, pharmaceutical, medical, and chemical have been well-defined. On the other hand, PHB is a promising raw material to produce biodegradable plastics. Although it was shown in our previous studies that H. smyrnensis AAD6T exhibits one of the highest conversion yields of sucrose to levan reported to date, novel strategies are required to overcome high costs of levan production. In this study, we aimed at increasing levan productivity of H. smyrnensis AAD6T cultures using random mutagenesis techniques combined (i.e., ethyl methanesulfate treatment and/or ultraviolet irradiation). After several consecutive treatments, mutant strains BAE2, BAE5 and BAE6 were selected as efficient levan producers, as BAE2 standing out as the most efficient one not only in sucrose utilization and levan production rates, but also in final PHB concentrations. The mutants' whole genome sequences were analysed to determine the mutations occurred. Several mutations in genes related to central carbon metabolism and osmoregulation were found. Our results suggest that random mutagenesis can be a facile and efficient strategy to enhance the performance of extremophiles in adverse conditions. • Random mutagenesis can be a facile and efficient strategy to enhance the performance of extremophiles in adverse conditions. • An improved levan producer Halomonas strain is obtained, characterized and described. • Sucrose utilization and levan production rates, as well as final PHB yields were enhanced. [ABSTRACT FROM AUTHOR]

Extreme ecosystems such as hypersaline habitats are promising source of bacteria able to produce potent natural molecules. Halophilic and halotolerant bacteria associated with an unexplored dry salt lake in Algeria have been investigated. A total of 17 isolates were phenotypically characterized. Optima temperature, pH, and salt concentration for growth have been determined. Bacteria strains have been identified based on the 16S rRNA gene sequencing. Enzymes production and antimicrobial activity against bacteria and fungi strains have been investigated. The results showed that isolates were assigned to three phylotypes, within eight isolates belonging to the genus Virgibacillus, eight isolates to Bacillus, and the genus Halomonas with one representative isolate. All phylotypes were able to thrive on large scales of salt, temperature and pH. In addition, the screening of extracellular enzymes showed that all the isolates were able to produce at least two enzymes. The gelatinase, and caseinase activities were the most produced. The screening of antimicrobial compounds production revealed that these strains exhibited significant activity against Gram-positive, Candida albicans, Fusarium, and Penicillium species. Finally, this biotechnological profiling and the PCA analysis have been shown that LMB3038, LMB3095, and LMB3082 were the most performant strains and present a promising source of important molecules. [ABSTRACT FROM AUTHOR]

Halobacterium sp. strain NMX12-1, an extremely halophilic Archaeon, was isolated from 250 million-year-old Salado Formation salt crystal in Carlsbad, New Mexico. Single-molecule real-time sequencing revealed a 3.2-Mbp genome with a 2.6-Mbp chromosome and five plasmids (234, 211, 119, 21, and 1.6-kbp). The GC-rich genome encodes an acidic proteome, characteristic of Haloarchaea., Competing Interests: R.J.R. and T.V. work at, and B.P.A. formerly worked for, New England Biolabs, a company that sells research reagents, including restriction enzymes and DNA methyltransferases, to the scientific community. None of the other authors declare any conflict of interest.

Terrestrial mud volcanoes represent surface features of channels for subsurface methane transport and, therefore, constitute an important source of methane emission from natural environments. How microbial processes regulate methane emissions in terrestrial mud volcanoes has yet to be fully addressed. This study demonstrated the geochemical characteristics and microbial communities of four mud volcano and seep sites in two geological settings of Sicily, Italy. At sites within the accretionary wedge that exhibited higher methane and sulfate concentrations, the communities were dominated by members capable of catalyzing methane and sulfate metabolisms and organic degradation. In particular, both anaerobic and aerobic methanotrophs were abundant and their abundance distribution coincided with the geochemical transition. In contrast, the sites near Mount Etna were characterized by high fluid salinity, CO 2 , and low methane and sulfate concentrations, with communities consisting of halophilic organic degraders and sulfur metabolizers, along with a minor presence of aerobic methanotrophs. Substantial variations in community composition and geochemistry across spatial and vertical redox gradients suggest that physicochemical contexts imposed by the geology, fluid path, and source characteristics play a vital role in shaping community composition and cycling of methane, sulfur and organic carbon in Sicily mud volcanoes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Chen, Chiu, Tu, Italiano, Wang and Lin.)

Halophilic microbial eukaryotes are present in many eukaryotic lineages and major groups; however, our knowledge of their diversity is still limited. Furthermore, almost nothing is known about the intracellular accumulation of salts in most halophilic eukaryotes. Here, we isolate a novel halophilic microbial eukaryote from hypersaline water of 134 practical salinity units (PSU) in a solar saltern. This species is an amoeboflagellate (capable of the amoeba-flagellate-cyst transformation) in the heterolobosean group and belongs to the genus Euplaesiobystra based on morphological data and 18S rDNA sequences. However, the isolate is distinct from any of the described Euplaesiobystra species. Especially, it is the smallest Euplaesiobystra to date, has a distinct cytostome, and grows optimally at 75-100 PSU. Furthermore, the phylogenetic tree of the 18S rDNA sequences demonstrates that the isolate forms a strongly supported group, sister to Euplaesiobystra hypersalinica. Thus, we propose that the isolate, Euplaesiobystra salpumilio, is a novel species. E. salpumilio displays a significantly increased influx of the intracellular Na+ and K+ at 50, 100, and 150 PSU, compared to freshwater species. However, the intracellular retention of the Na+ and K+ at 150 PSU does not significantly differ from 100 PSU, suggesting that E. salpumilio can extrude the Na+ and K+ from cells under high-salinity conditions. Interestingly, actively growing E. salpumilio at 100 and 150 PSU may require more intracellular accumulation of Na+ than the no-growth but-viable state at 50 PSU. It seems that our isolate displays two salt metabolisms depending on the tested salinities. E. salpumilio shows a salt-in strategy for Na+ at lower salinity of 100 PSU, while it displays a salt-out strategy for Na+ at higher salinity of 150 PSU. Our results suggest that the novel halophilic E. salpumilio fundamentally uses a salt-out strategy at higher salinities, and the accumulation patterns of intracellular salts in this species are different from those in other halophilic microbial eukaryotes. [ABSTRACT FROM AUTHOR]

The use of wild plant species or their halophytic relatives has been considered in plant breeding programs to improve salt and drought tolerance in crop plants. Aeluropus littoralis serves as halophyte model for identification and isolation of novel stress adaptation genes. A. littoralis, a perennial monocot grass, grows in damp or arid areas, often salt-impregnated places and wasteland in cultivated areas, can survive periodically high water salinity, and tolerate high salt concentrations in the soil up to 1,100 mM sodium chloride. Therefore, it serves as valuable genetic resource to understand molecular mechanisms of stress-responses in monocots. The knowledge can potentially be used for improving tolerance to abiotic stresses in economically important crops. Several morphological, anatomical, ecological, and physiological traits of A. littoralis have been investigated so far. After watering with salt water the grass is able to excrete salt via its salt glands. Meanwhile, a number of ESTs (expressed sequence tag), genes and promoters induced by the salt and drought stresses were isolated, sequenced and annotated at a molecular level. Transfer of stress related genes to other species resulted in enhanced stress resistance. Here we describe the genome sequence and structure of A. littoralis analyzed by whole genome sequencing and histological analysis. The chromosome number was determined to be 20 (2n = 2x = 20). The genomesize wascalculated to be 354 Mb. This genomic information provided here, will support the functional investigation and application of novel genes improving salt stress resistance in crop plants. The utility of the sequence information is exemplified by the analysis of the DREB-transcription factor family. [ABSTRACT FROM AUTHOR]

Phenanthrene (Phe), a tricyclic Polycyclic Aromatic Hydrocarbon (PAH), is found in high concentrations as a pollutant in various environments. In this study, the removal or (oxidizing) ability of Phe by a laccase from Alkalibacillus almallahensis was investigated. The laccase (12 U mL−1) was able to remove 63% of Phe (50 mg L−1) under optimal conditions of 40 °C, pH 8, 1.5 M NaCl and in the presence of 1 mM HBT as a laccase mediator after a 72 h incubation period. The results for the effect of different solvents, ionic and non-ionic surfactants on the activity of the halophilic laccase towards Phe showed that the addition of these compounds increase removal efficiency and complete enzymatic removal of Phe will achieve in a solution of 5% (v/v) acetone and 1.5% tween 80. The kinetic parameters Km and Vmax of laccase-catalyzed removal of the substrate were determined as 0.544 mM and 0.882 µmol h−1 mg−1, respectively. A microtoxicity study with respect to the inhibition of algal growth showed a decrease in toxicity of the laccase-treated Phe solution. [ABSTRACT FROM AUTHOR]

Halophilic phage are a type of virus that exist in salty environments within halophilic archaeal or bacterial hosts. However, relatively few reports on halophilic bacteriophages exist, and our overall understanding of halophilic bacteriophages is quite limited. We used SYBR Green I fluorescent staining to detect the abundance of viruses in Yuncheng Saline Lake, China. Using the double-layer plate method, a lytic phage that could infect halophilic bacterium Salinivibrio sp. YM-43 was isolated and named YXM43. We studied host range, optimal host, morphological characteristics, nucleic acid type, protein composition, and other biological characteristics of the virus. Results reveal a high abundance of this halophilic virus in Yuncheng Saline Lake. The newly isolated bacteriophage YXM43 has a narrow host range, with the most suitable host being Virgibacillus sp. SK39. After purification and enrichment, YXM43 is observed as a spherical particle with a diameter of approximately 30 nm, with no tail. No lipid envelope can be seen in YXM43. The capsid protein of the virus can be separated into seven proteins with molecular weights ranging from 62.0 to 13.0 kDa. YXM43 is a DNA virus with a genome approximately 23 kb. The virus is tolerant of low salinity, and its activity is highest at a temperature of 60 °C and a pH of 10. YXM43 is temperature and pH tolerant, and can adapt to environmental change, even withstanding chloroform treatment. The results indicate that bacteriophage YXM43 is a novel halophilic bacteriophage with broad tolerance to environmental change. [ABSTRACT FROM AUTHOR]

Abstract Background Lentibacillus species are gram variable aerobic bacteria that live primarily in halophilic environments. Previous reports have shown that bacteria belonging to this species are primarily isolated from salty environments or food. We isolated a bacterial strain CBA3610, identified as a novel species of the genus Lentibacillus, from a human fecal sample. In this report, the whole genome sequence of Lentibacillus sp. CBA3610 is presented, and genomic analyses are performed. Results Complete genome sequence of strain CBA3610 was obtained through PacBio RSII and Illumina HiSeq platforms. The size of genome is 4,035,571 bp and genes estimated to be 4714 coding DNA sequences and 64 tRNA and 17 rRNA were identified. The phylogenetic analysis confirmed that it belongs to the genus Lentibacillus. In addition, there were genes related to antibiotic resistance and virulence, and genes predicted as CRISPR and prophage were also identified. Genes related to osmotic stress were found according to the characteristics of halophilic bacterium. Genomic differences from other Lentibacillus species were also confirmed through comparative genomic analysis. Conclusions Strain CBA3610 is predicted to be a novel candidate species of Lentibacillus through phylogenetic analysis and comparative genomic analysis with other species in the same genus. This strain has antibiotic resistance gene and pathogenic genes. In future, the information derived from the results of several genomic analyses of this strain is thought to be helpful in identifying the relationship between halophilic bacteria and human gut microbiota.

Extremophilic organisms demonstrate the flexibility and adaptability of basic biological processes by highlighting how cell physiology adapts to environmental extremes. Few eukaryotic extremophiles have been well studied and only a small number are amenable to laboratory cultivation and manipulation. A detailed characterization of the genome architecture of such organisms is important to illuminate how they adapt to environmental stresses. One excellent example of a fungal extremophile is the halophile Hortaea werneckii (Pezizomycotina, Dothideomycetes, Capnodiales), a yeast-like fungus able to thrive at near-saturating concentrations of sodium chloride and which is also tolerant to both UV irradiation and desiccation. Given its unique lifestyle and its remarkably recent whole genome duplication, H. werneckii provides opportunities for testing the role of genome duplications and adaptability to extreme environments. We previously assembled the genome of H. werneckii using short-read sequencing technology and found a remarkable degree of gene duplication. Technology limitations, however, precluded high-confidence annotation of the entire genome. We therefore revisited the H. wernickii genome using long-read, single-molecule sequencing and provide an improved genome assembly which, combined with transcriptome and nucleosome analysis, provides a useful resource for fungal halophile genomics. Remarkably, the ∼50 Mb H. wernickii genome contains 15,974 genes of which 95% (7608) are duplicates formed by a recent whole genome duplication (WGD), with an average of 5% protein sequence divergence between them. We found that the WGD is extraordinarily recent, and compared to Saccharomyces cerevisiae, the majority of the genome's ohnologs have not diverged at the level of gene expression of chromatin structure.

The use of wild plant species or their halophytic relatives has been considered in plant breeding programs to improve salt and drought tolerance in crop plants. Aeluropus littoralis serves as halophyte model for identification and isolation of novel stress adaptation genes. A. littoralis, a perennial monocot grass, grows in damp or arid areas, often salt-impregnated places and wasteland in cultivated areas, can survive periodically high water salinity, and tolerate high salt concentrations in the soil up to 1,100 mM sodium chloride. Therefore, it serves as valuable genetic resource to understand molecular mechanisms of stress-responses in monocots. The knowledge can potentially be used for improving tolerance to abiotic stresses in economically important crops. Several morphological, anatomical, ecological, and physiological traits of A. littoralis have been investigated so far. After watering with salt water the grass is able to excrete salt via its salt glands. Meanwhile, a number of ESTs (expressed sequence tag), genes and promoters induced by the salt and drought stresses were isolated, sequenced and annotated at a molecular level. Transfer of stress related genes to other species resulted in enhanced stress resistance. Here we describe the genome sequence and structure of A. littoralis analyzed by whole genome sequencing and histological analysis. The chromosome number was determined to be 20 (2n = 2x = 20). The genome size was calculated to be 354 Mb. This genomic information provided here, will support the functional investigation and application of novel genes improving salt stress resistance in crop plants. The utility of the sequence information is exemplified by the analysis of the DREB-transcription factor family.

Biologically enhanced transport of radionuclides is one of several processes that can affect the performance of a nuclear waste repository. In this work, several microbial isolates from the Waste Isolation Pilot Plant (WIPP) were tested for their influence on the concentration of neodymium, as an analog for +3 actinides, in simple sodium chloride solutions and in anoxic WIPP brines. Batch sorption experiments were carried out over a period of 4–5 weeks. In many cases, the effect on neodymium in solution was immediate and extensive and assumed to be due to surface complexation. However, over time, the continued loss of Nd from the solution was more likely due to biologically induced precipitation and/or mineralization and possible entrapment in extracellular polymeric substances. The results showed no correlation between organism type and the extent of its influence on neodymium in solution. However, a correlation was observed between different test matrices (simple NaCl versus high-magnesium brine versus high-NaCl brine). Further experiments were conducted to test these matrix effects, and the results showed a significant effect of magnesium concentration on the ability of microorganisms to remove Nd from solution. Possible mechanisms include cation competition and the alteration of cell surface structures. This suggests that the aqueous chemistry of the WIPP environs could play a larger role in the final disposition of +3 actinides than the microbiology.

Halophilic bacteria such as the genus Halomonas are promising candidates in diverse industrial, agricultural and biomedical applications. Here, we successfully isolated a halophilic Halomonas alkaliphila strain XH26 from Xiaochaidan Salt Lake, and studied its osmoadaptation strategies using transcriptome and ectoine analysis. Divergent mechanisms were involved in osmoadaptation at different salinities in H. alkaliphila XH26. At moderate salinity (6% NaCl), increased transcriptions of ABC transporters related to iron (III), phosphate, phosphonate, monosaccharide and oligosaccharide import were observed. At high salinity (15% NaCl), transcriptions of flagellum assembly and cell motility were significantly inhibited. The transcriptional levels of ABC transporter genes related to iron (III) and iron3+-hydroxamate import, glycine betaine and putrescine uptake, and cytochrome biogenesis and assembly were significantly up-regulated. Ectoine synthesis and accumulation was significantly increased under salt stress, and the increased transcriptional expressions of ectoine synthesis genes ectB and ectC may play a key role in high salinity induced osmoadaptation. At extreme high salinity (18% NaCl), 5-hydroxyectoine and ectoine worked together to maintain cell survival. Together these results give valuable insights into the osmoadaptation mechanisms of H. alkaliphila XH26, and provide useful information for further engineering this specific strain for increased ectoine synthesis and related applications. [ABSTRACT FROM AUTHOR]

Adaption to environmental conditions is reflected by protein adaptation. In particular, proteins of extremophiles display distinctive traits ensuring functional, structural and dynamical properties under permanently extreme physical and chemical conditions. While it has mostly been studied with approaches focusing on specific proteins, biophysical approaches have also confirmed this link between environmental and protein adaptation at the more complex and diverse scale of the proteome. However, studies of this type remain challenging and often require large amounts of biological material. We report here the use of nanoDSF as a tool to study proteome stability and solubility in cell lysates of the model halophilic archaeon Haloarcula marismortui. Notably, our results show that, as with single halophilic protein studies, proteome stability was correlated to the concentration of NaCl or KCl under which the cells were lysed and hence the proteome exposed. This work highlights that adaptation to environmental conditions can be experimentally observed at the scale of the proteome. Still, we show that the biochemical properties of single halophilic proteins can only be partially extrapolated to the whole proteome. [ABSTRACT FROM AUTHOR]