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Great Salt Lake (GSL), located northwest of Salt Lake City, UT, is the largest terminal lake in the USA. While the average salinity of seawater is ~3.3%, the salinity in GSL ranges between 5% and 28%. In addition to being a hypersaline environment, GSL also contains toxic concentrations of heavy metals, such as arsenic, mercury, and lead. The extreme environment of GSL makes it an intriguing subject of study, both for its unique microbiome and its potential to harbor novel natural product-producing bacteria, which could be used as resources for the discovery of biologically active compounds. Though work has been done to survey and catalog bacteria found in GSL, the Lakes microbiome is largely unexplored, and little to no work has been done to characterize the natural product potential of GSL microbes. Here, we investigate the bacterial diversity of two important regions within GSL, describe the first genomic characterization of Actinomycetota isolated from GSL sediment, including the identification of two new Actinomycetota species, and provide the first survey of the natural product potential of GSL bacteria.

Strain MP-1014T, an obligate halophilic actinobacterium, was isolated from the mangrove soil of Thandavarayancholanganpettai, Tamil Nadu, India. A polyphasic approach was utilized to explore its phylogenetic position completely. The isolate was Gram-positive, filamentous, non-motile, and coccoid in older cultures. Ideal growth conditions were seen at 30 °C and pH 7.0, with 5% NaCl (W/V), and the DNA G + C content was 73.3%. The phylogenic analysis of this strain based upon 16S rRNA gene sequence revealed 97–99.8% similarity to the recognized species of the genus Isoptericola. Strain MP-1014T exhibits the highest similarity to I. sediminis JC619T (99.7%), I. chiayiensis KCTC19740T (98.9%), and subsequently to I. halotolerans KCTC19646T (98.6%), when compared with other members within the Isoptericola genus (< 98%). ANI scores of strain MP-1014T are 86.4%, 84.2%, and 81.5% and dDDH values are 59.7%, 53.6%, and 34.8% with I. sediminis JC619T, I. chiayiensis KCTC19740T and I. halotolerans KCTC19646T respectively. The major polar lipids of the strain MP-1014T were phosphatidylinositol, phosphatidylglycerol, diphosphotidylglycerol, two unknown phospholipids, and glycolipids. The predominant respiratory menaquinones were MK9 (H4) and MK9 (H2). The major fatty acids were anteiso-C15:0, anteiso-C17:0, iso-C14:0, C15:0, and C16:0. Also, initial genome analysis of the organism suggests it as a biostimulant for enhancing agriculture in saline environments. Based on phenotypic and genetic distinctiveness, the strain MP-1014 T represents the novel species of the genus Isoptericola assigned Isoptericola haloaureus sp. nov., is addressed by the strain MP-1014 T, given its phenotypic, phylogenetic, and hereditary uniqueness. The type strain is MP-1014T [(NCBI = OP672482.1 = GCA_036689775.1) ATCC = BAA-2688T; DSMZ = 29325T; MTCC = 13246T]. [ABSTRACT FROM AUTHOR]

Hypersaline ecosystems are distributed all over the globe. They are subjected to poly-extreme stresses and are inhabited by halophilic microorganisms possessing multiple adaptations. The halophiles have many biotechnological applications such as nutrient supplements, antioxidant synthesis, salt tolerant enzyme production, osmolyte synthesis, biofuel production, electricity generation etc. However, halophiles are still underexplored in terms of complex ecological interactions and functions as compared to other niches. The advent of metagenomics and the recent advancement of next-generation sequencing tools have made it feasible to investigate the microflora of an ecosystem, its interactions and functions. Both target gene and shotgun metagenomic approaches are commonly employed for the taxonomic, phylogenetic, and functional analyses of the hypersaline microbial communities. This review discusses different types of hypersaline niches, their residential microflora, and an overview of the metagenomic approaches used to investigate them. Various applications, hurdles and the recent advancements in metagenomic approaches have also been focused on here for their better understanding and utilization in the study of hypersaline microbiome. [ABSTRACT FROM AUTHOR]

Extreme environments, such as highly saline ecosystems, are characterised by a limited presence of microbial communities capable of tolerating and thriving under these conditions. To better understand the limits of life and its chemical and microbiological drivers, highly saline and brine groundwaters of Na‐Cl and Na‐Ca‐Cl types with notably diverse SO4 contents were sampled in water intakes and springs from sedimentary aquifers located in the Outer Carpathians and the Carpathian Foredeep basin and its basement in Poland. Chemical and microbiological methods were used to identify the composition of groundwaters, determine microbial diversity, and indicate processes controlling their distribution using multivariate statistical analyses. DNA sequencing targeting V3‐V4 and V4‐V5 gene regions revealed a predominance of Proteobacteriota, Methanobacteria, Methanomicrobia, and Nanoarchaea in most of the water samples, irrespective of their geological context. Despite the sample‐size constraint, redundancy analysis employing a compositional approach to hydrochemical predictors identified Cl/SO4 and Cl/HCO3 ratios, and specific electrical conductivity, as key gradients shaping microbial communities, depending on the analysed gene regions. Analysis of functional groups revealed that methanogenesis, sulphate oxidation and reduction, and the nitrogen cycle define and distinguish the halotolerant communities in the samples. These communities are characterised by an inverse relationship between methanogens and sulphur‐cycling microorganisms. [ABSTRACT FROM AUTHOR]

Abstract Halophiles are one of the classes of extremophilic microorganisms that can flourish in environments with very high salt concentrations. In this study, fifteen bacterial strains isolated from various crop rhizospheric soils of agricultural fields along the Southwest coastline of Saurashtra, Gujarat, and identified by 16S rRNA gene sequencing as Halomonas pacifica, H. stenophila, H. salifodinae, H. binhaiensis, Oceanobacillus oncorhynchi, and Bacillus paralicheniformis were investigated for their potentiality to produce extremozymes and compatible solute. The isolates showed the production of halophilic protease, cellulase, and chitinase enzymes ranging from 6.90 to 35.38, 0.004–0.042, and 0.097–0.550 U ml−1, respectively. The production of ectoine-compatible solute ranged from 0.01 to 3.17 mg l−1. Furthermore, the investigation of the ectoine-compatible solute production at the molecular level by PCR showed the presence of the ectoine synthase gene responsible for its biosynthesis in the isolates. Besides, it also showed the presence of glycine betaine biosynthetic gene betaine aldehyde dehydrogenase in the isolates. The compatible solute production by these isolates may be linked to their ability to produce extremozymes under saline conditions, which could protect them from salt-induced denaturation, potentially enhancing their stability and activity. This correlation warrants further investigation.

Ectoine, a biologically significant compound, was successfully produced by a strain of bacteria capable of utilizing sucrose. In a ground-breaking approach, we harnessed the potential of sugar beet molasses, a by-product rich in sucrose, amino acid, and vitamins, as a growth medium for this purpose. Through meticulous investigation, we identified the ideal conditions for maximizing ectoine synthesis. This remarkable milestone was reached by introducing only 1 g of (NH₄)₂SO₄ and 5 mL of molasses per liter, maintaining a pH level of 8.0, upholding a 7.5% NaCl concentration, employing agitation at 120 rpm, and sustaining a temperature of 30 °C. This study marks a pioneering endeavour as it represents the first instance where molasses has been effectively employed to produce ectoine through the cultivation of Nesterenkonia sp. We showcased the production of 75.56 g of the valuable compound ectoine utilizing 1 L of waste molasses with this specific bacterial strain. These findings hold tremendous promise, not only in terms of resource utilization but also for the potential applications of ectoine in various biological contexts. [ABSTRACT FROM AUTHOR]

Azo dye-laden textile wastewater must be treated before release due to various health and environmental concerns. Bioremediation of textile wastewater, however, is a challenge owing to its alkaline and saline nature as mesophilic microbes, in general, are either not able to thrive or show less efficiency under such hostile environment. Thus, pre-treatment for neutralization or salinity removal becomes a prerequisite before applying microbes for treatment, causing extra economical and technical burden. Extremophilic bacteria can be the promising bioremediating tool because of their inherent ability to survive and show toxicants removal capability under such extreme conditions without need of pre-treatment. Among extremophiles, halophilic and alkaliphilic bacteria which are naturally adapted to high salt and pH are of special interest for the decolorization of saline-alkaline-rich textile wastewater. The current review article is an attempt to provide an overview of the bioremediation of azo dyes and azo dye-laden textile wastewater using these two classes of extremophilic bacteria. The harmful effects of azo dyes on human health and environment have been discussed herein. Halo-alkaliphilic bacteria circumvent the extreme conditions by various adaptations, e.g., production of certain enzymes, adjustment at the protein level, pH homeostasis, and other structural adaptations that have been highlighted in this review. The unique properties of alkaliphiles and halophiles, to not only sustain but also harboring high dye removal competence at high pH and salt concentration, make them a good candidate for designing future bioremediation strategies for the management of alkaline, salt, and azo dye-laden industrial wastewaters. [ABSTRACT FROM AUTHOR]

Halophiles are one of the classes of extremophilic microorganisms that can flourish in environments with very high salt concentrations. In this study, fifteen bacterial strains isolated from various crop rhizospheric soils of agricultural fields along the Southwest coastline of Saurashtra, Gujarat, and identified by 16S rRNA gene sequencing as Halomonas pacifica, H. stenophila, H. salifodinae, H. binhaiensis, Oceanobacillus oncorhynchi, and Bacillus paralicheniformis were investigated for their potentiality to produce extremozymes and compatible solute. The isolates showed the production of halophilic protease, cellulase, and chitinase enzymes ranging from 6.90 to 35.38, 0.004–0.042, and 0.097–0.550 U ml−1, respectively. The production of ectoine-compatible solute ranged from 0.01 to 3.17 mg l−1. Furthermore, the investigation of the ectoine-compatible solute production at the molecular level by PCR showed the presence of the ectoine synthase gene responsible for its biosynthesis in the isolates. Besides, it also showed the presence of glycine betaine biosynthetic gene betaine aldehyde dehydrogenase in the isolates. The compatible solute production by these isolates may be linked to their ability to produce extremozymes under saline conditions, which could protect them from salt-induced denaturation, potentially enhancing their stability and activity. This correlation warrants further investigation. [ABSTRACT FROM AUTHOR]

Extremophilic proteins are valuable in various fields, but their expression can be challenging in traditional hosts like Escherichia coli due to misfolding and aggregation. Haloferax volcanii (H. volcanii), a halophilic expression system, offers a solution. This study examined cleavable and non-cleavable purification tags at both the N- and C-termini when fused with the superfolder green fluorescent protein (sfGFP) in H. volcanii. Our findings reveal that an N-terminal 8xHis-tag or Strep-tag®II significantly enhances protein production, purity, and yield in H. volcanii. Further experiments with mCherry and halophilic alcohol dehydrogenase (ADH) showed improved expression and purification yields when the 8xHis-tag or Strep-tag®II was positioned at the C-terminus for mCherry and at the N-terminus for ADH. Co-positioning 8xHis-tag and Twin-Strep-tag® at the N-terminus of sfGFP, mCherry, and ADH yielded significantly enhanced results. These findings highlight the importance of thoughtful purification tag design and selection in H. volcanii, providing valuable insights for improving protein production and purification with the potential to advance biotechnological applications. [ABSTRACT FROM AUTHOR]

Extremophile organisms have adapted to extreme physicochemical conditions. Halophilic organisms, in particular, survive at very high salt concentrations. To achieve this, they have engineered the surface of their proteins to increase the number of short, polar and acidic amino acids, while decreasing large, hydrophobic and basic residues. While these adaptations initially decrease protein stability in the absence of salt, they grant halophilic proteins remarkable stability in environments with extremely high salt concentrations, where non‐adapted proteins unfold and aggregate. The molecular mechanisms by which halophilic proteins achieve this, however, are not yet clear. Here, we test the hypothesis that the halophilic amino acid composition destabilizes the surface of the protein, but in exchange improves the stability in the presence of salts. To do that, we have measured the folding thermodynamics of various protein variants with different degrees of halophilicity in the absence and presence of different salts, and at different pH values to tune the ionization state of the acidic amino acids. Our results show that halophilic amino acids decrease the stability of halophilic proteins under mesophilic conditions, but in exchange improve salt‐induced stabilization and solubility. We also find that, in contrast to traditional assumptions, contributions arising from hydrophobic effect and preferential ion exclusion are more relevant for haloadaptation than electrostatics. Overall, our findings suggest a trade‐off between folding thermodynamics and halophilic adaptation to optimize proteins for hypersaline environments. [ABSTRACT FROM AUTHOR]

Background: Hypersaline environments (> 40 practical salinity units [PSU]) represent some of the most extreme conditions on Earth, supporting a variety of halophilic and halotolerant bacteria, archaea, and protists. The taxon Heterolobosea includes numerous halophilic protists, making it a valuable model for studying eukaryotic adaptation to high salinity. Particularly, the genus Pharyngomonas, a deep-branching lineage within Heterolobosea, comprises mainly obligate halophiles, providing insights into early protist adaptations in hypersaline environments. Additionally, these protozoa play crucial ecological roles as grazers of bacteria and archaea, and are prey for higher trophic levels in hypersaline environments. Results: In the present study, two previously reported amoeboflagellates were isolated for the first time from hypersaline waters (~300 PSU) in two solar salterns in the Republic of Korea. Microscopic observations revealed that both strains exhibited the characteristic morphologies of Pharyngomonas, including amoeboid, flagellate, and cyst forms. Molecular phylogenetic analysis of their 18S rRNA gene sequences confirmed their close relationship to known Pharyngomonas kirbyi strains. The two strains demonstrated growth within a salinity range of 75–200 PSU, with optimal growth observed at 75–100 PSU, confirming their status as true halophiles. All known P. kirbyi strains are obligate halophiles, exhibiting a clear instance of adaptive radiation of halophilic eukaryotes. Additionally, the genus Pharyngomonas has been found in hypersaline environments across multiple continents (Asia, Europe, North America, Australia, and Africa), suggesting that it plays an ecologically significant role as a grazer of prokaryotes or prey for higher trophic levels in these habitats. Conclusions: On the bases of morphological and molecular analyses, two strains identified as P. kirbyi were isolated and characterized for the first time from solar salterns in the Republic of Korea. This discovery highlights the presence and adaptation of halophilic eukaryotes in such extreme environments. The confirmation of these strains as obligate halophiles provides additional evidence for the adaptive radiation of halophilic eukaryotes. Furthermore, the ecological role of Pharyngomonas species underscores their importance as trophic regulators in hypersaline ecosystems. These findings contribute to a deeper understanding of the diversity, adaptation, and ecological functions of halophilic eukaryotes in extreme environments.

The Dead Sea is unique compared to other extreme halophilic habitats. Its salinity exceeds 34%, and it is getting saltier. The Dead Sea environment is characterized by a dominance of divalent cations, with magnesium chloride (MgCl2) levels approaching the predicted 2.3 M upper limit for life, an acidic pH of 6.0, and high levels of absorbed ultraviolet radiation. Consequently, only organisms adapted to such a polyextreme environment can survive in the surface, sinkholes, sediments, muds, and underwater springs of the Dead Sea. Metagenomic sequence analysis and amino acid profiling indicated that the Dead Sea is predominantly composed of halophiles that have various adaptation mechanisms and produce metabolites that can be utilized for biotechnological purposes. A variety of products have been obtained from halophilic microorganisms isolated from the Dead Sea, such as antimicrobials, bioplastics, biofuels, extremozymes, retinal proteins, colored pigments, exopolysaccharides, and compatible solutes. These resources find applications in agriculture, food, biofuel production, industry, and bioremediation for the detoxification of wastewater and soil. Utilizing halophiles as a bioprocessing platform offers advantages such as reduced energy consumption, decreased freshwater demand, minimized capital investment, and continuous production.

Microbial natural products from microbes in extreme environments, including haloarchaea, and halophilic bacteria, possess a huge capacity to produce novel antibiotics. Additionally, enhanced isolation techniques and improved tools for genomic mining have expanded the efficiencies in the antibiotic discovery process. This review article provides a detailed overview of known antimicrobial compounds produced by halophiles from all three domains of life. We summarize that while halophilic bacteria, in particular actinomycetes, contribute the vast majority of these compounds the importance of understudied halophiles from other domains of life requires additional consideration. Finally, we conclude by discussing upcoming technologies- enhanced isolation and metagenomic screening, as tools that will be required to overcome the barriers to antimicrobial drug discovery. This review highlights the potential of these microbes from extreme environments, and their importance to the wider scientific community, with the hope of provoking discussion and collaborations within halophile biodiscovery. Importantly, we emphasize the importance of bioprospecting from communities of lesser-studied halophilic and halotolerant microorganisms as sources of novel therapeutically relevant chemical diversity to combat the high rediscovery rates. The complexity of halophiles will necessitate a multitude of scientific disciplines to unravel their potential and therefore this review reflects these research communities. [ABSTRACT FROM AUTHOR]

Toluene and other monoaromatic compounds are released into the environment particularly saline habitats due to the inappropriate disposal methods of petroleum products. Studying the bio-removal strategy is required to clean up these hazardous hydrocarbons that threaten all ecosystem life using halophilic bacteria with higher biodegradation efficiency of monoaromatic compounds as a sole carbon and energy source. Therefore, sixteen pure halophilic bacterial isolates were obtained from saline soil of Wadi An Natrun, Egypt, which have the ability to degrade toluene and consume it as the only source of carbon and energy. Amongst these isolates, isolate M7 exhibited the best growth with considerable properties. This isolate was selected as the most potent strain and identified based on phenotypic and genotypic characterizations. The strain M7 was belonging to Exiguobacterium genus and founded to be closely matched to the Exiguobacterium mexicanum with a similarity of 99%. Using toluene as sole carbon source, strain M7 showed good growth at a wide range temperature degree (20–40ºC), pH (5–9), and salt concentrations (2.5–10%, w/v) with optimal growth conditions at 35ºC, pH 8, and 5%, respectively. The biodegradation ratio of toluene was estimated at above optimal conditions and analyzed using Purge-Trap GC-MS. The results showed that strain M7 has the potentiality to degraded 88.32% of toluene within greatly short time (48 h). The current study findings support the potential ability to use strain M7 as a biotechnological tool in many applications such as effluent treatment and toluene waste management. [ABSTRACT FROM AUTHOR]

The nitrogen discharge from saline wastewater will cause significant pollution to the environment. As a high-efficiency and low-cost treatment method, biological treatment has a promising application prospect in the removal of nitrogen from high-salt wastewater. However, the inhibitory effect of high salt on microorganisms increases the difficulty of its treatment. This review discusses the influence of salinity on the nitrogen removal process, considering both traditional and novel biological techniques. Common methods to enhance the effectiveness of biological nitrogen removal processes and their mechanisms of action in engineering practice and research, including sludge acclimation and inoculation of halophilic bacteria, are also introduced. An outlook on the future development of biological nitrogen removal processes for high-salt wastewater is provided to achieve environmentally friendly discharge of high-salt wastewater. [ABSTRACT FROM AUTHOR]

The given document is a list of various scientific articles and research papers related to topics such as Martian mineralogy, the habitability of Mars and Europa, the survival of bacteria in Martian conditions, the adaptation mechanisms of halophilic archaea, and the analysis of gene expression in different organisms. The articles cover a range of subjects including astrobiology, RNA sequencing, osmotic stress, and the effects of environmental factors on microbial life. The document provides a comprehensive collection of scientific literature that can be useful for library patrons conducting research on these specific topics. [Extracted from the article]

Six soil samples from three layers of an archaeological investigation profile from a pre-industrial ash deposit place have been investigated by NGS analyses of 16 S rRNA. The three pairs of sample originate from top soil (internal reference), from an intermediate ash layer and from a lower ash layer, formed about two centuries ago. In addition to general abundant bacteria, special genera known as halophilic or alkaline-tolerant have been found as expected from the history of the place and from the measured pH-value and conductivity measurements. The close relations between samples of pairs and the differences between the three soil layers are clearly indicated by abundance correlation and PCA-diagrams. Comparative PCA correlation plots including samples from an archaeological excavation site dedicated to pre-industrial coal mining illustrate the high distinguishability of investigated soils. These relations are particular clearly shown when lower abundant bacteria are regarded. The investigations are a further example for the "ecological memory of soil" reflecting the strong human impact on this pre-industrial embossed place. [ABSTRACT FROM AUTHOR]

Acidic salt lakes are environments that harbor an array of biologically challenging conditions. Through 16S rRNA, 18S rRNA, and ITS amplicon sequencing of eight such lakes across the Yilgarn Craton of Western Australia, we aim to understand the microbial ecology of these lakes with a focus on iron- and sulfur-oxidizing and reducing microorganisms that have theoretical application in biomining industries. In spite of the biological challenges to life in these lakes, the microbial communities were highly diverse. Redundancy analysis of soil samples revealed sulfur, ammonium, organic carbon, and potassium were significant diversities of the microbial community composition. The most abundant microbes with a hypothetical application in biomining include the genus 9  M32 of the Acidithiobacillus family, Alicyclobacillus and Acidiphilium, all of which are possible iron- and/or sulfur-oxidizing bacteria. It is evident through this study that these lakes harbor multiple organisms with potential in biomining industries that should be exploited and studied further. [ABSTRACT FROM AUTHOR]

Recent geological, hydrochemical, and mineralogical studies performed on hypersaline salt flats have given insights into similar geo-morphologic features on Mars. These salt-encrusted depressions are widely spread across the Earth, where they are characterized by high salt concentrations, intense UV radiation, high evaporation, and low precipitation. Their surfaces are completely dry in summer; intermittent flooding occurs in winter turning them into transitory hypersaline lakes. Thanks to new approaches such as culture-dependent, culture-independent, and metagenomic-based methods, it is important to study microbial life under polyextreme conditions and understand what lives in these dynamic ecosystems and how they function. Regarding these particular features, new halophilic microorganisms have been isolated from some salt flats and identified as excellent producers of primary and secondary metabolites and granules such as halocins, enzymes, carotenoids, polyhydroxyalkanoates, and exopolysaccharides. Additionally, halophilic microorganisms are implemented in heavy metal bioremediation and hypersaline wastewater treatment. As a result, there is a growing interest in the distribution of halophilic microorganisms around the world that can be looked upon as good models to develop sustainable biotechnological processes for all fields. This review provides insights into diversity, ecology, metabolism, and genomics of halophiles in hypersaline salt flats worldwide as well as their potential uses in biotechnology. [ABSTRACT FROM AUTHOR]

AbstractThe heterocyclic ether 1,4-Dioxane (1,4-DE) is a toxic by-product used widely in various industrial activities as a solvent or stabilizer. 1,4-DE was classified as a probable human carcinogen according to the United States Environmental Protection Agency (USEPA) due to its chemical and physical properties. Emerging pollutants in industrial wastewater is the major challenge for the operation of effluent treatment plant. The bioremediation of 1,4-DE is an effective approach to treat the industrial wastewater. The bacterial consortium was obtained from industrial sludge samples, Yanbu, Saudi Arabia and enriched in 1,4-DE supported basal salt saline medium. Biodegradation of 1,4-DE was studied at several concentrations ranging from 200 to 1200 mg/L. Complete mineralization of 1,4-DE from 200 to 800 mg/L was achieved in 4 to 8 days at 4% salinity. 1,4-DE utilization was studied at different salinity (4% and 6%), the results exhibited complete mineralization of 1,4-DE at 4 days and 6 days respectively. Increase in salinity to 8% displayed 92% degradation of 1,4-DE. Addition of yeast extract accelerated the biomass production correspondingly reduced the time required for biodegradation under saline condition. The bacterial consortium showed the ability to degrade 1,4-DE completely at various range of concentrations. Lab scale continuous stirred tank reactor (CSTR) was used to evaluate the industrial wastewater treatment efficacy of the bacterial consortium under saline condition. CSTR operated for 36 days displayed chemical oxygen demand (COD) removal and 1,4-DE degradation of 96% under saline condition. THF (Tetrahydrofuran) which accelerated the degradation process of 1,4-DE was completely degraded in 28 days. Bacterial analysis with high throughput sequencing technique confirmed the existence of bacterial strains like Ochrobactrum (45%), Marinobacter (21%), Stenotrophomonas (16%), Pseuodomonas (9%), Sphingomonas (4%), Bacillus (3.5%) and Sedimentibacter (1.5%). [ABSTRACT FROM AUTHOR]

Microbial syntrophy, a cooperative metabolic interaction among prokaryotes, serves a critical role in shaping communities, due to the auxotrophic nature of many microorganisms. Syntrophy played a key role in the evolution of life, including the hypothesized origin of eukaryotes. In a recent exploration of the microbial mats within the exceptional and uniquely extreme Cuatro Cienegas Basin (CCB), a halophilic isolate, designated as AD140, emerged as a standout due to its distinct growth pattern. Subsequent genome sequencing revealed AD140 to be a co-culture of a halophilic archaeon from the Halorubrum genus and a marine halophilic bacterium, Marinococcus luteus, both occupying the same ecological niche. This intriguing coexistence hints at an early-stage symbiotic relationship that thrives on adaptability. By delving into their metabolic interdependence through genomic analysis, this study aims to uncover shared characteristics that enhance their symbiotic association, offering insights into the evolution of halophilic microorganisms and their remarkable adaptations to high-salinity environments. [ABSTRACT FROM AUTHOR]

Aim The present study aims to determine the antimicrobial potential of Virgibacillus salairus (MML1918) against human pathogens and its in-vitro and in-silico properties. Methods and results In this present study, totally 63 halophilic bacterial cultures were obtained and cultivated in nutrient broth medium containing 8% NaCl and the metabolites, were extracted using ethyl acetate and screened for their antimicrobial property by cell viability assay against 12 pathogenic bacteria and fungi, among 63 halophilic bacteria the Vir. salaries (MML1918) found to be the best producer for secondary metabolites production against clinical pathogens. The optimization of growth for important physiochemical parameters was characterized and applied for different production media and based on its highest activity as 17.5 ± .07 mm zone of inhibition (ZOI) for Bacillus cereus followed by 17.5 ± 00 mm ZOI for Staphylococcus aureus , the production medium ATCC1097 was chosen for mass production. The mass production of secondary metabolites from Vir. salaries MML1918 was carried out in a fermenter under controlled conditions and crude metabolites was extracted and condensed. The antimicrobial activity of crude metabolites showed B. cereus (19.3 ± 0.5 mm ZOI), Staph. aureus , and Candida albicans (18.3 ± 0.5 mm ZOI) as the highest ZOI in production media for halophilic bacteria ATCC1097. Further, the gas chromatography–mass spectrometry analysis showed 24 compounds present in crude metabolites. Among the 24 compounds, four molecules were found to be important based on molecule percentage in crude and structural similarity. The molecular docking studies show that the selected four molecules effectively bind with the active region DNA gyrase B. Conclusion Virgibacillus salarius (MML1918) effectively showed antimicrobial activity against several pathogenic organisms and can be employed as a suitable candidate for producing novel antimicrobial agents. [ABSTRACT FROM AUTHOR]

Microbial cellulases are an important industrial enzyme having diverse applications in biotechnology, environmental challenges, industrial products and processes. Extremophiles like thermophillic bacteria are a good source of industrially important cellulases as these can withstand industrially rigorous procedures like paper deinking, fabric material softening, bio stoning, paper and pulp, biopolishing cloth material, animal feed and juice. Identification of novel cellulases or improving them through biotechnological interventions has remained a challenge for researchers. Genetic manipulation of thermophilic bacteria for increased cellulase production or synthetic biology approaches for cellulase gene/gene cluster extraction from thermophilic bacteria and expression in appropriate hosts for improved cellulase synthesis. The classic and high -throughput technologies like genomics, metagenomics and bioinformatics could be exploited to isolate cellulase genes from a variety of thermophilic bacteria and further processing. Keeping in view the ultimate requirement of extremophilic cellulases in industries, present study is a compilation of various aspects related to extremophilic cellulases their sources, production, biotechnological interventions and challenges. [ABSTRACT FROM AUTHOR]

Carotenoids are one of the most essential groups of naturally occurring lipid-soluble pigments that display great biological activities, such as antioxidant, anti-inflammatory, and provitamin A activities. Carotenoids are widely applied in various industries due to their advantageous health effects. Microbial carotenoid synthesis, therefore, has attracted increasing attention in recent years. In the present study, halophiles isolated and characterised from the Solar saltern of Mulund, Mumbai, Maharashtra, were determined to be a strain of G. terrae based on its 16S rRNA gene sequence. The strain G. terrae has the ability to synthesise and accumulate the intracellular pigments, which was identified by gas chromatography-mass spectrometry (GC-MS) and Fourier transform infrared (FTIR). Findings suggested that G. terrae can actively grow and efficiently synthesise carotenoids. GC-MS results revealed that the pigment sample contains nearly 17 different compounds. 17 recorded compounds are terpenoids and organic pigments. These compounds were recorded for the first time from the Solar saltern of Mulund, Mumbai, Maharashtra. Therefore, the new bacterial strain isolated and characterised from the Solar saltern of Mulund, Mumbai, showed a high potential bioresource for the commercial production of natural carotenoids. [ABSTRACT FROM AUTHOR]

The current study presents an overview of heavy metals bioremediation from halo–alkaline conditions by using extremophilic microorganisms. Heavy metal remediation from the extreme environment with high pH and elevated salt concentration is a challenge as mesophilic microorganisms are unable to thrive under these polyextremophilic conditions. Thus, for effective bioremediation of extreme systems, specialized microbes (extremophiles) are projected as potential bioremediating agents, that not only thrive under such extreme conditions but are also capable of remediating heavy metals from these environments. The physiological versatility of extremophiles especially halophiles and alkaliphiles and their enzymes (extremozymes) could conveniently be harnessed to remediate and detoxify heavy metals from the high alkaline saline environment. Bibliometric analysis has shown that research in this direction has found pace in recent years and thus this review is a timely attempt to highlight the importance of halo-alkaliphiles for effective contaminant removal in extreme conditions. Also, this review systematically presents insights on adaptive measures utilized by extremophiles to cope with harsh environments and outlines the role of extremophilic microbes in industrial wastewater treatment and recovery of metals from waste with relevant examples. Further, the major challenges and way forward for the effective applicability of halo-alkaliphilic microbes in heavy metals bioremediation from extremophilic conditions are also highlighted. [ABSTRACT FROM AUTHOR]

The family Halomonadaceae is the largest family composed of halophilic bacteria, with more than 160 species with validly published names as of July 2023. Several classifications to circumscribe this family are available in major resources, such as those provided by the List of Prokaryotic names with Standing in Nomenclature (LPSN), NCBI Taxonomy, Genome Taxonomy Database (GTDB), and Bergey's Manual of Systematics of Archaea and Bacteria (BMSAB), with some degree of disagreement between them. Moreover, regardless of the classification adopted, the genus Halomonas is not phylogenetically consistent, likely because it has been used as a catch-all for newly described species within the family Halomonadaceae that could not be clearly accommodated in other Halomonadaceae genera. In the past decade, some taxonomic rearrangements have been conducted on the Halomonadaceae based on ribosomal and alternative single-copy housekeeping gene sequence analysis. High-throughput technologies have enabled access to the genome sequences of many type strains belonging to the family Halomonadaceae; however, genome-based studies specifically addressing its taxonomic status have not been performed to date. In this study, we accomplished the genome sequencing of 17 missing type strains of Halomonadaceae species that, together with other publicly available genome sequences, allowed us to re-evaluate the genetic relationship, phylogeny, and taxonomy of the species and genera within this family. The approach followed included the estimate of the Overall Genome Relatedness Indexes (OGRIs) such as the average amino acid identity (AAI), phylogenomic reconstructions using amino acid substitution matrices customized for the family Halomonadaceae, and the analysis of clade-specific signature genes. Based on our results, we conclude that the genus Halovibrio is obviously out of place within the family Halomonadaceae, and, on the other hand, we propose a division of the genus Halomonas into seven separate genera and the transfer of seven species from Halomonas to the genus Modicisalibacter, together with the emendation of the latter. Additionally, data from this study demonstrate the existence of various synonym species names in this family. [ABSTRACT FROM AUTHOR]

Extremophilic proteins are valuable in various fields, but their expression can be challenging in traditional hosts like Escherichia coli due to misfolding and aggregation. Haloferax volcanii (H. volcanii), a halophilic expression system, offers a solution. This study examined cleavable and non-cleavable purification tags at both the N- and C-termini when fused with the superfolder green fluorescent protein (sfGFP) in H. volcanii. Our findings reveal that an N-terminal 8xHis-tag or Strep-tag®II significantly enhances protein production, purity, and yield in H. volcanii. Further experiments with mCherry and halophilic alcohol dehydrogenase (ADH) showed improved expression and purification yields when the 8xHis-tag or Strep-tag®II was positioned at the C-terminus for mCherry and at the N-terminus for ADH. Co-positioning 8xHis-tag and Twin-Strep-tag® at the N-terminus of sfGFP, mCherry, and ADH yielded significantly enhanced results. These findings highlight the importance of thoughtful purification tag design and selection in H. volcanii, providing valuable insights for improving protein production and purification with the potential to advance biotechnological applications.

The production of second-generation biofuel requires a huge amount of freshwater. It is estimated that at least three gal of freshwater is used to produce one gal of biofuel. The replacement of freshwater with seawater serves as a potential alternative in biofuel generation. Therefore, salt-tolerant enzymes play an important role in saccharification and fermentation process. Halophilic β-glucosidase is one of the key enzymes for the process. In this study, the β-glucosidase of halophile Meridianimaribacter sp. CL38 isolated from mangrove soil was characterized. Strain CL38 achieved maximum production of β-glucosidase at 12th hour of growth. The β-glucosidase showed highest activity at 2% (w/v) NaCl while highly stable at salt concentration ranging from 1-2% (w/v) (more than 96% of relative activity). Its β-glucosidase activity remained active in the presence of 5mM Mn2+, Mg2+, Ca2+ ions, and 1% (v/v) Tween-20 and Tween-80. The draft genome sequence of strain CL38 was retrieved from GenBank database and submitted to dbCAN meta server for CAZymes annotation. Strain CL38 harbors 44 GHs and GH3 are annotated as β-glucosidases. The β-glucosidases of Meridianimaribacter flavus (99.61%) and Mesoflavibacter sabulilitoris (97.44%) showed the closest identity with Bgl3a and Bgl3b protein sequences from strain CL38, respectively. Glycoside hydrolase family 3 domain was identified in both enzymes via InterPro scan server. The presence of signal peptides indicated that both enzymes were secreted extracellularly. Five motifs were identified in Bgl3a and Bgl3b, with the active site (nucleophile) found at Asp296 and Asp297, respectively. Collectively, these β-glucosidases could be potentially used in the biofuel production, in particular the lignocellulosic biomass pretreatment process. This is the first attempt to characterize the β-glucosidase in genus Meridianimaribacter as so far none of the lignocellulolytic enzymes from this genus were characterized.

Abstract Background Ovarian cancer is one of the most frequent and deadly gynaecological cancers, often resistant to platinum-based chemotherapy, the current standard of care. Halophilic microorganisms have been shown to produce a large variety of metabolites, some of which show toxicity to various cancer cell lines. However, none have yet been shown to be active against ovarian cancer cells. Here, we examined the effects of metabolites secreted by the halophilic archaea Halorhabdus rudnickae and Natrinema salaciae on various cancer cell lines, including ovarian cancer cell lines. Results 1H NMR analyses of Hrd. rudnickae and Nnm. salaciae culture supernatants contain a complex mixture of metabolites that differ between species, and even between two different strains of the same species, such as Hrd. rudnickae strains 64T and 66. By using the MTT and the xCELLigence RTCA assays, we found that the secreted metabolites of all three halophilic strains expressed cytotoxicity to the ovarian cancer cell lines, especially A2780, as well as its cisplatin-resistant derivative A2780cis, in a dose-dependent manner. The other tested cell lines A549, HepG2, SK-OV-3 and HeLa were only minimally, or not at all affected by the archaeal metabolites, and this was only seen with the MTT assay. Conclusions The halophilic archaea Hrd. rudnickae and Nnm. salaciae, isolated from a Polish salt mine and Lake Medee in the Mediterranean Sea, respectively, secrete metabolites that are active against ovarian cancer cells, including those that are resistant to cisplatin. This opens potential new possibilities for the treatment of these frequent and deadly gynaecological cancers.

Acidic salt lakes are environments that harbor an array of biologically challenging conditions. Through 16S rRNA, 18S rRNA, and ITS amplicon sequencing of eight such lakes across the Yilgarn Craton of Western Australia, we aim to understand the microbial ecology of these lakes with a focus on iron- and sulfur-oxidizing and reducing microorganisms that have theoretical application in biomining industries. In spite of the biological challenges to life in these lakes, the microbial communities were highly diverse. Redundancy analysis of soil samples revealed sulfur, ammonium, organic carbon, and potassium were significant diversities of the microbial community composition. The most abundant microbes with a hypothetical application in biomining include the genus 9 M32 of the Acidithiobacillus family, Alicyclobacillus and Acidiphilium, all of which are possible iron- and/or sulfur-oxidizing bacteria. It is evident through this study that these lakes harbor multiple organisms with potential in biomining industries that should be exploited and studied further.

Microbial syntrophy, a cooperative metabolic interaction among prokaryotes, serves a critical role in shaping communities, due to the auxotrophic nature of many microorganisms. Syntrophy played a key role in the evolution of life, including the hypothesized origin of eukaryotes. In a recent exploration of the microbial mats within the exceptional and uniquely extreme Cuatro Cienegas Basin (CCB), a halophilic isolate, designated as AD140, emerged as a standout due to its distinct growth pattern. Subsequent genome sequencing revealed AD140 to be a co-culture of a halophilic archaeon from the Halorubrum genus and a marine halophilic bacterium, Marinococcus luteus, both occupying the same ecological niche. This intriguing coexistence hints at an early-stage symbiotic relationship that thrives on adaptability. By delving into their metabolic interdependence through genomic analysis, this study aims to uncover shared characteristics that enhance their symbiotic association, offering insights into the evolution of halophilic microorganisms and their remarkable adaptations to high-salinity environments.

Introduction: This work investigates whether rhizosphere microorganisms that colonize halophyte plants thriving in saline habitats can tolerate salinity and provide beneficial effects to their hosts, protecting them from environmental stresses, such as aromatic compound (AC) pollution. Methods: To address this question, we conducted a series of experiments. First, we evaluated the effects of phenol, tyrosine, 4-hydroxybenzoic acid, and 2,4-dichlorophenoxyacetic (2,4-D) acids on the soil rhizosphere microbial community associated with the halophyte Allenrolfea vaginata. We then determined the ability of bacterial isolates from these microbial communities to utilize these ACs as carbon sources. Finally, we assessed their ability to promote plant growth under saline conditions. Results: Our study revealed that each AC had a different impact on the structure and alpha and beta diversity of the halophyte bacterial (but not archaeal) communities. Notably, 2,4-D and phenol, to a lesser degree, had the most substantial decreasing effects. The removal of ACs by the rhizosphere community varied from 15% (2,4-D) to 100% (the other three ACs), depending on the concentration. Halomonas isolates were the most abundant and diverse strains capable of degrading the ACs, with strains of Marinobacter, Alkalihalobacillus, Thalassobacillus, Oceanobacillus, and the archaea Haladaptatus also exhibiting catabolic properties. Moreover, our study found that halophile strains Halomonas sp. LV-8T and Marinobacter sp. LV-48T enhanced the growth and protection of Arabidopsis thaliana plants by 30% to 55% under salt-stress conditions. Discussion: These results suggest that moderate halophile microbial communities may protect halophytes from salinity and potential adverse effects of aromatic compounds through depurative processes. [ABSTRACT FROM AUTHOR]

Soil salinity is one of the most important factors reducing agricultural productivity worldwide. Halophilic plant growth-promoting bacteria (H-PGPB) represent an alternative method of alleviating saline stress in crops of agricultural interest. In this study, the following halophilic bacteria were evaluated: Bacillus sp. SVHM1.1, Halomonas sp. SVCN6, Halomonas sp. SVHM8, and a consortium. They were grown under greenhouse conditions in Solanum lycopersicum at different salinity concentrations in irrigation water (0, 20, 60, and 100 mM NaCl) to determine the effects on germination, fruit quality, yield, and concentration of osmoprotectors in plant tissue. Our results demonstrate the influence of halophilic bacteria with the capacity to promote plant growth on the germination and development of Solanum lycopersicum at higher salinity levels. The germination percentage was improved at the highest concentration by the inoculated treatments (from 37 to 47%), as were the length of the radicle (30% at 20 mM) and plumule of the germinated seed, this bacterium also increased the weight of the plumule (97% at 100 mM). They also improved the yield. The dry weight of the plant, in addition to having an influence on the quality of the fruit and the concentration of osmoprotectors (Bacillus sp. SVHM 1.1) had the greatest effect on fruit yield (1.5 kg/plant at 20 mM), by the otherhand, Halomonas sp. SVHM8 provided the best fruit quality characteristics at 100 mM. According to the above results, the efficiency of halophilic PGPB in the attenuation of salt stress in Solanum lycopersicum has been proven. [ABSTRACT FROM AUTHOR]

The olive oil industry faces a major problem of treating the wastewater with high organic content and safe disposal. Olive oil industrial wastewater (OOIWW) consists of highly toxic environmental pollutants with high salinity. Saline olive oil industrial wastewater was treated using halophilic consortium in UMFC (upflow microbial fuel cell) mobilized with carbon felt as electrode. Total and soluble COD (chemical oxygen demand), total suspended solids and phenol content removal were studied at different organic loads (0.56, 0.77, 1.05, 1.26, 1.52 and 1.8 gCOD/L). UMFC with OOIWW was optimized at 1.52 gCOD/L for high organic removal and corresponding electricity production. Total COD, soluble COD, TSS and phenol removal were 91%, 89%, 78%, and complete removal of phenol was accomplished at the optimized organic load (1.52 gCOD/L). Correspondingly, the maximum bioenergy yield was 784 mV with 439 mW/m2 (power density) and 560 mA/m2 (current density), respectively. The presence of prominent halophilic exo-electrogens such as Ochrobactrum, Marinobacter, Rhodococcus and Bacillus potently treated the OOIWW and exhibited high energy yield. [ABSTRACT FROM AUTHOR]

Simple Summary: A significant amount of residual oil is located in oil reservoirs with high-salinity formation water levels. Microorganisms are capable of producing a number of oil-displacing metabolites, such as volatile acids, alcohols, biosurfactants, biopolymers, and others. The application of microbial-enhanced oil recovery (MEOR) methods is possible after a detailed study of the microbial community of the oil reservoir. The aim of this study was to elucidate the biodiversity of microorganisms in oil reservoirs of Tatarstan (Russia) with high saline formation water in order to select suitable MEOR technologies for these reservoirs. Using metagenomic and cultural methods, it was shown that fermentative and sulfate-reducing bacteria predominate in reservoirs with high-sulfate formation water, whereas in oil reservoirs with low-sulfate water, in the community of Methanohalophilus methanogens predominated. A total of 20 pure bacterial cultures were isolated from oil reservoirs. The isolated fermentative bacteria were able to produce oil-displacing metabolites from sugar-containing and protein substrates. However, fermentation products stimulate the growth of sulfidogenic bacteria that form sulfide, which reduces the quality of oil and causes the corrosion of steel equipment. Thus, when choosing MEOR technology, it is necessary to take into account the possibility of activation of fermentative prokaryotes with simultaneous suppression of the growth of sulfidogens in the oil reservoir. The reserves of light conditional oil in reservoirs with low-salinity formation water are decreasing worldwide, necessitating the extraction of heavy oil from petroleum reservoirs with high-salinity formation water. As the first stage of defining the microbial-enhanced oil recovery (MEOR) strategies for depleted petroleum reservoirs, microbial community composition was studied for petroleum reservoirs with high-salinity formation water located in Tatarstan (Russia) using metagenomic and culture-based approaches. Bacteria of the phyla Desulfobacterota, Halanaerobiaeota, Sinergistota, Pseudomonadota, and Bacillota were revealed using 16S rRNA-based high-throughput sequencing in halophilic microbial communities. Sulfidogenic bacteria predominated in the studied oil fields. The 75 metagenome-assembled genomes (MAGs) of prokaryotes reconstructed from water samples were assigned to 16 bacterial phyla, including Desulfobacterota, Bacillota, Pseudomonadota, Thermotogota, Actinobacteriota, Spirochaetota, and Patescibacteria, and to archaea of the phylum Halobacteriota (genus Methanohalophilus). Results of metagenomic analyses were supported by the isolation of 20 pure cultures of the genera Desulfoplanes, Halanaerobium, Geotoga, Sphaerochaeta, Tangfeifania, and Bacillus. The isolated halophilic fermentative bacteria produced oil-displacing metabolites (lower fatty acids, alcohols, and gases) from sugar-containing and proteinaceous substrates, which testify their potential for MEOR. However, organic substrates stimulated the growth of sulfidogenic bacteria, in addition to fermenters. Methods for enhanced oil recovery should therefore be developed, combining the production of oil-displacing compounds with fermentative bacteria and the suppression of sulfidogenesis. [ABSTRACT FROM AUTHOR]

Halophilic extracellular proteases offer promising application in various fields. Information on these prominent proteins including the synthesizing organisms, biochemical properties, domain organisation, purification, and application challenges has never been covered in recent reviews. Although extracellular proteases from bacteria pioneered the study of proteases in halophiles, progress is being made in proteases from halophilic archaea. Recent advances in extracellular proteases from archaea revealed that archaeal proteases are more robust and applicable. Extracellular proteases are composed of domains that determine their mechanisms of action. The intriguing domain structure of halophilic extracellular proteases consists of N-terminal domain, catalytic domain, and C-terminal extension. The role of C-terminal domains varies among different organisms. A high diversity of C-terminal domains would endow the proteases with diverse functions. With the development of genomics, culture-independent methods involving heterologous expression, affinity chromatography, and in vitro refolding are deployed with few challenges on purification and presenting novel research opportunities. Halophilic extracellular proteases have demonstrated remarkable potentials in industries such as detergent, leather, peptide synthesis, and biodegradation, with desirable properties and ability to withstand harsh industrial processes. Key points: • Halophilic extracellular proteases have robust properties suitable for applications. • A high diversity of C-terminal domains may endow proteases with diverse properties. • Novel protease extraction methods present novel application opportunities. [ABSTRACT FROM AUTHOR]

Extremophiles are the most fascinating life forms for their special adaptations and ability to offer unique extremozymes or bioactive molecules. Halophiles, the natural inhabitants of hypersaline environments, are one among them. Halomonas are the common genus of halophilic bacteria. To support growth in unusual environments, Halomonas produces various hydrolytic enzymes, compatible solutes, biopolymers like extracellular polysaccharides (EPS) and polyhydroxy alkaloates (PHA), antibiotics, biosurfactants, pigments, etc. Many of such molecules are being produced in large-scale bioreactors for commercial use. However, the prospect of the remaining bioactive molecules with industrial relevance is far from their application. Furthermore, the genetic engineering of the respective gene clusters could open up a new path to bio-prospect these molecules by overproducing their products through heterologous expression. The present survey on Halomonas highlights their ecological diversity, application potential of the their various industrially relevant biomolecules and impact of these biomolecules on respective fields. [ABSTRACT FROM AUTHOR]

Production of insensitive munitions (IM) creates wastewater containing 3-nitro-1,2,4-triazol-5-one (NTO). Because NTO is an anion in neutral pH wastewaters, it can be removed via ion exchange (IX), released during IX brine recharge, and may be subsequently degraded by halophiles. IX brine regeneration through halophilic NTO biotransformation was observed over 210 cycles in a sequencing batch reactor (SBR) and averaged 64.8%±20.2% NTO removal. NTO formed 3-amino-1,2,4-triazol-5-one (ATO) under sulfate- and nitrate-reducing conditions. Variable NTO reductions were observed over four phases including anoxic enrichment (80.5%±7.7% , Cycles 0 to 41); anoxic followed by aerobic (42.2%±23.5% , Cycles 42 to 81); anoxic supplemented with nitrate (64.7%±21.5% , Cycles 82 to 177); and anoxic stress test with nitrate and perchlorate (91.0%±9.7% , Cycles 178 to 210). Sequencing results showed 69% of the genera to be halophiles and sulfate-reducing bacteria and archaea including Desulfovibrio and Halanaerobium. ATO reduction in each cycle was shown to be correlated with the abundance of genus Alkalibacterium. The presence of the potential cocontaminants in munitions wastewater IX brine, nitrate and perchlorate, were shown to either increase or not interfere with NTO transformation, respectively. The results of these studies demonstrate that NTO biotransformation by halophiles in brine solutions is feasible and could be utilized for IX brine regeneration. Explosive production and munitions manufacturing in the US creates wastewater containing explosive contaminants. Treatment of this wastewater to remove the explosives is required before the water can be released to the environment. IX can remove the explosive NTO from munitions manufacturing wastewater, but to be economical, the resins need to be reused many times. Regeneration of the IX resin for reuse generates brine that contains the explosive NTO. To remove NTO from the brine so it can be reused, salt-tolerant bacteria were grown that biodegrade NTO. The bacteria were able to biotransform NTO over for 210 cycles and over 500 days from a synthetic IX brine. After optimization of the treatment system, the bacteria were degrading 91.0%± 9.7% of the NTO in each cycle. This system, IX with brine regeneration, for munitions wastewater treatment is potentially more economical and sustainable than current wastewater treatment methods. [ABSTRACT FROM AUTHOR]

The contribution of halophilic microorganisms to the quality of salts produced in solar salterns has recently been recognized but not clearly understood. Using metagenomic 16S rRNA gene sequencing approach, we showed the microbial composition difference between the Tuban crystallization pond (CP-Tuban) that produces low-quality salt and CP-Sampang as a representative solar saltern that yields high-quality salt. Dominant classes in both traditional salterns were represented by γ-proteobacteria and halobacteria that occurred at higher prevalence in CP-Sampang. Microbial taxa, including beneficial genera, in CP-Sampang were more diverse and abundant compared to CP-Tuban. Among 180 genus-level OTUs identified in CP-Sampang, 127 of them were considered unique due to their absence in CP-Tuban. Higher levels of dissolved oxygen (DO) and nutrient (phosphate, nitrate, and ammonia) in the seawater reservoir (SR) of Sampang may contribute to more diverse phytoplankton genera, which could support the growth of beneficial heterotrophic microbes that positively affect the salt quality of the CP-Sampang. Low number of Dunaleilla sp. in both CPs do not seem to influence the quality of salts produced. The outcome of these comparative studies provides new insights into the contribution of diverse microbial taxa in correlation with physico-chemical parameters and phytoplankton communities to the high quality of salts produced in traditional solar salterns. The presence of beneficial genera in the enriched microbial cultures could provide an important basis for further applications, such as improving the quality of salt produced and producing unique compounds and enzymes. [ABSTRACT FROM AUTHOR]

The production of second-generation biofuel requires a huge amount of freshwater. It is estimated that at least three gal of freshwater is used to produce one gal of biofuel. The replacement of freshwater with seawater serves as a potential alternative in biofuel generation. Therefore, salt-tolerant enzymes play an important role in saccharification and fermentation process. Halophilic β-glucosidase is one of the key enzymes for the process. In this study, the β-glucosidase of halophile Meridianimaribacter sp. CL38 isolated from mangrove soil was characterized. Strain CL38 achieved maximum production of β-glucosidase at 12th hour of growth. The β-glucosidase showed highest activity at 2% (w/v) NaCl while highly stable at salt concentration ranging from 1-2% (w/v) (more than 96% of relative activity). Its β-glucosidase activity remained active in the presence of 5mM Mn2+, Mg2+, Ca2+ ions, and 1% (v/v) Tween-20 and Tween-80. The draft genome sequence of strain CL38 was retrieved from GenBank database and submitted to dbCAN meta server for CA-Zymes annotation. Strain CL38 harbors 44 GHs and GH3 are annotated as β-glucosidases. The β-glucosidases of Meridianimaribacter flavus (99.61%) and Mesoflavibacter sabulilitoris (97.44%) showed the closest identity with Bgl3a and Bgl3b protein sequences from strain CL38, respectively. Glycoside hydrolase family 3 domain was identified in both enzymes via InterPro scan server. The presence of signal peptides indicated that both enzymes were secreted extracellularly. Five motifs were identified in Bgl3a and Bgl3b, with the active site (nucleophile) found at Asp296 and Asp297, respectively. Collectively, these β-glucosidases could be potentially used in the biofuel production, in particular the lignocellulosic biomass pretreatment process. This is the first attempt to characterize the β-glucosidase in genus Meridianimaribacter as so far none of the lignocellulolytic enzymes from this genus were characterized. [ABSTRACT FROM AUTHOR]

Abstract Background Various bacteria and archaea, including halophilic archaeon Halobacterium sp. NRC-1 produce gas vesicle nanoparticles (GVNPs), a unique class of stable, air-filled intracellular proteinaceous nanostructures. GVNPs are an attractive tool for biotechnological applications due to their readily production, purification, and unique physical properties. GVNPs are spindle- or cylinder-shaped, typically with a length of 100 nm to 1.5 μm and a width of 30–250 nm. Multiple monomeric subunits of GvpA and GvpC proteins form the GVNP shell, and several additional proteins are required as minor structural or assembly proteins. The haloarchaeal genetic system has been successfully used to produce and bioengineer GVNPs by fusing several foreign proteins with GvpC and has shown various applications, such as biocatalysis, diagnostics, bioimaging, drug delivery, and vaccine development. Results We demonstrated that native GvpC can be removed in a low salt buffer during the GVNP purification, leaving the GvpA-based GVNP's shell intact and stable under physiological conditions. Here, we report a genetic engineering and chemical modification approach for functionalizing the major GVNP protein, GvpA. This novel approach is based on combinatorial cysteine mutagenesis within GvpA and genetic expansion of the N-terminal and C-terminal regions. Consequently, we generated GvpA single, double, and triple cysteine variant libraries and investigated the impact of mutations on the structure and physical shape of the GVNPs formed. We used a thiol–maleimide chemistry strategy to introduce the biotechnological relevant activity by maleimide-activated streptavidin–biotin and maleimide-activated SpyTag003-SpyCatcher003 mediated functionalization of GVNPs. Conclusion The merger of these genetic and chemical functionalization approaches significantly extends these novel protein nanomaterials' bioengineering and functionalization potential to assemble catalytically active proteins, biomaterials, and vaccines onto one nanoparticle in a modular fashion. Graphical Abstract

The family Halomonadaceae is the largest family composed of halophilic bacteria, with more than 160 species with validly published names as of July 2023. Several classifications to circumscribe this family are available in major resources, such as those provided by the List of Prokaryotic names with Standing in Nomenclature (LPSN), NCBI Taxonomy, Genome Taxonomy Database (GTDB), and Bergey’s Manual of Systematics of Archaea and Bacteria (BMSAB), with some degree of disagreement between them. Moreover, regardless of the classification adopted, the genus Halomonas is not phylogenetically consistent, likely because it has been used as a catch-all for newly described species within the family Halomonadaceae that could not be clearly accommodated in other Halomonadaceae genera. In the past decade, some taxonomic rearrangements have been conducted on the Halomonadaceae based on ribosomal and alternative single-copy housekeeping gene sequence analysis. High-throughput technologies have enabled access to the genome sequences of many type strains belonging to the family Halomonadaceae; however, genome-based studies specifically addressing its taxonomic status have not been performed to date. In this study, we accomplished the genome sequencing of 17 missing type strains of Halomonadaceae species that, together with other publicly available genome sequences, allowed us to re-evaluate the genetic relationship, phylogeny, and taxonomy of the species and genera within this family. The approach followed included the estimate of the Overall Genome Relatedness Indexes (OGRIs) such as the average amino acid identity (AAI), phylogenomic reconstructions using amino acid substitution matrices customized for the family Halomonadaceae, and the analysis of clade-specific signature genes. Based on our results, we conclude that the genus Halovibrio is obviously out of place within the family Halomonadaceae, and, on the other hand, we propose a division of the genus Halomonas into seven separate genera and the transfer of seven species from Halomonas to the genus Modicisalibacter, together with the emendation of the latter. Additionally, data from this study demonstrate the existence of various synonym species names in this family.