Extremophiles最新文献

Early characterizations by morphological identification through light microscopy only revealed the presence of a few microbial lineages and the majority of microbial community at the Chae Son hot spring remains uncharacterized. Therefore, this study aims to examine thermophilic microbial communities at the Chae Son hot spring using next-generation sequencing, including investigating hot spring mineralogy. Results suggest that the Chae Son hot spring (49-75 °C, pH = 6.5-7.0) precipitates digitate structures which comprise mainly silica, and that microbial permineralization is primarily through silicification. Alternating layers of mineralized microbial biofilms and silica were observed in digitate sinter cross-sections, contributing to the build-up of microstromatolites. Molecular results revealed that phylogenetically distinct members of photoautotrophic taxa, Chloroflexota and Cyanobacteriota, dominated spring microbial communities (63.19% relative abundance). Potential primary production processes were mainly through photoautotrophy, with minor lithoautotrophic activities (e.g., sulfur cycling and nitrogen cycling). Moreover, overall microbial community and Cyanobacteriota population alpha diversities significantly decreased with increased temperatures. However, no significant correlation was identified between Chloroflexota population diversity and temperatures. This study provides an update on the microbial community using a high-throughput next-generation sequencing technology, including the mineralogy of the Chae Son hot spring, Northern Thailand.

The cell membrane remodeling mediated by cyclopropane fatty acid synthase (CfaS) plays a crucial role in microbial physiological processes resisting various environmental stressors, including acid. Herein, we found a relatively high proportion (24.8%-28.3%) of cyclopropane fatty acid (CFA) Cy-19:0 in the cell membrane of a newly isolated extreme acidophile, Acidithiobacillus caldus CCTCC AB 2019256, under extreme acid stress. Overexpression of the CfaS encoding gene cfaS2 in Escherichia coli conferred enhanced acid resistance. GC-MS analysis revealed a 3.52-fold increase in the relative proportion of Cy-19:0 in the cell membrane of the overexpression strain compared to the control. Correspondingly, membrane fluidity, permeability and cell surface hydrophobicity were reduced to varying degrees. Additionally, HPLC analysis indicated that the overexpression strain had 1.54-, 1.42-, 1.85-, 1.20- and 1.05-fold higher levels of intracellular glutamic acid, arginine, aspartic acid, methionine and alanine, respectively, compared to the control. Overall, our findings shed light on the role of CfaS derived from extreme acidophile in bacterial defense against environmental acid stress, potentially facilitating its application in the design and development of industrial microbial chassis cells for organic acid production.

A novel extremely halophilic archaeon designated, MBLA0158^T, was isolated from a solar saltern in Sorae, Republic of Korea. The colonies are red-pigmented, Gram-stain-negative, pleomorphic, non-motile, and lysed in distilled water. The strain grows at 25-45 °C (optimum, 37 °C), in 15-30% (w/v) NaCl (optimum, 20%) and 0.1-1.0 M Mg²⁺ (optimum, 0.2-0.3 M) at pH 6.0-10.0 (optimum, 7.0-8.0). Comparative analysis based on the 16S rRNA gene sequence revealed that this strain is most closely related to the Halobellus inordinatus YC20^T with a sequence identity of 96.0%. Strain MBLA0158^T contained phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester as major polar lipids. The genome size is 3.29 Mb and the DNA G + C content is 66.9 mol%. Phylogenomic analysis confirmed that strain MBLA0158^T is distinct from previously reported type strains of the genus Halobellus. Pan-genome analysis showed that strain MBLA0158^T contains 419 genes that are not present in other type strains of the genus Halobellus. Based on overall analyses, strain MBLA0158^T is considered to represent a new species of the genus Halobellus, for which the name Halobellus rubicundus sp. nov. is proposed. The type strain is MBLA0158^T (= KCTC 4318^T = JCM 36642^T).

The halophilic archaeon Haloarcula hispanica utilizes the sugar alcohols mannitol and sorbitol as carbon and energy sources. Genes, enzymes, and transcriptional regulators involved in uptake and degradation of these sugar alcohols were identified by growth experiments with deletion mutants and enzyme characterization. It is shown that both mannitol and sorbitol are taken up via a single ABC transporter of the CUT1 transporter family. Then, mannitol and sorbitol are oxidized to fructose by two distinct dehydrogenases. Fructose is further phosphorylated to fructose-1-phosphate by a haloarchaeal ketohexokinase, providing the first evidence for a physiological function of ketohexokinase in prokaryotes. Finally, fructose-1-phosphate is phosphorylated via fructose-1-phosphate kinase to fructose-1,6-bisphosphate, which is cleaved to triosephosphates by a Class I fructose-1,6-bisphosphate aldolase. Two distinct transcriptional regulators, acting as activators, have been identified: an IclR-like regulator involved in activating genes for sugar alcohol uptake and oxidation to fructose, and a GfcR-like regulator that likely activates genes involved in the degradation of fructose to pyruvate. This is the first comprehensive analysis of a sugar alcohol degradation pathway in Archaea.

The current representatives of the family Natrialbaceae within the class Halobacteria were subjected to phylogenetic, phylogenomic, and comparative genomic analyses. The current species of Halobiforma and Halomontanus were found to be related to those of Natronobacterium and Natronoglomus, respectively. According to the cutoff value of average amino acid identity (AAI) (≤ 76%) proposed to differentiate genera within the family Natrialbaceae, Halobiforma, and Natronoglomus should be merged with Natronobacterium and Halomontanus, respectively. Beyond these, four novel halophilic archaeal strains, CCL63^T, AD-5^T, CG52^T, and KLK7^T, isolated from three saline lakes and a saline-alkaline land in China, were simultaneously subjected to polyphasic classification. The phenotypic, phylogenetic, phylogenomic, and comparative genomic analyses indicated that strain CCL63^T (= CGMCC 1.18663^T = JCM 35096^T) represents a novel genus of the family Natrialbaceae, strains AD-5^T (= CGMCC 1.13783^T = JCM 33734^T) and CG52^T (= CGMCC 1.17139^T = JCM 34160^T) represent two novel species of the genus Natronococcus, and strain KLK7^T (= MCCC 4K00128^T = KCTC 4307^T) represents a novel species of Haloterrigena. Halovalidus salilacus gen. nov., sp. nov., Natronococcus wangiae sp. nov., Natronococcus zhouii sp. nov., and Haloterrigena salinisoli sp. nov. are further proposed based on these type strains accordingly.

We isolated and characterized the community of cultivable fungi associated with marine macroalgae present in the Magellan sub-Antarctic straits and the South Shetland Islands, Maritime Antarctica, and evaluated their production of bioactive metabolites. A total of 201 filamentous fungal isolates were obtained. The genera Antarctomyces, Pseudogymnoascus, Microdochium, Trichoderma, Cladosporium, Penicillium, Neoascochyta, Entomortierella and Linnemannia were associated with Antarctic macroalgae, with Neoascochyta paspali being the most abundant taxon. In contrast, 12 taxa representing Cadophora, Microdochium, Penicillium, Pseudogymnoascus were associated with macroalgae from the Magellan sub-Antarctic, with Penicillium dominating the assemblages. The diversity indices of the fungal communities associated with macroalgae in the two regions were similar. Among 177 fungal extracts assessed for metabolite production, 31 (17.5%) showed strong phytotoxic activity and 17 (9.6%) showed anti-Trypanosoma cruzi activity. Penicillium showed the highest phytotoxic and anti-Trypanosoma activity values. The detection of taxa in common between the polar and cold temperate zones reinforces the need for further investigations of the distribution of species in these distinct ecoregions. The detection of bioactive extracts produced particularly by Penicillium representatives reinforces the potential to obtain active molecules that can be explored as natural products or as sources of bioactive compounds with application in agriculture and biomedicine.

L-asparaginase (ASNase, E.C. 3.5.1.1) catalyzes the deamination of L-asparagine to L-aspartic acid and ammonia and is widely used in medicine to treat acute lymphocytic leukemia. It also has significant applications in the food industry by inhibiting acrylamide formation. In this study, we characterized a thermostable ASNase from the hyper thermophilic strain, Pyrococcus yayanosii CH1. The recombinant enzyme (PyASNase) exhibited maximal activity at pH 8.0 and 85 °C. Moreover, PyASNase demonstrated promising thermostability across temperatures ranging from 70 to 95 °C. The kinetic parameters of PyASNase for L-asparagine were a K_m of 6.3 mM, a k_cat of 1989s^-1, and a k_cat/K_m of 315.7 mM^-1 s^-1. Treating potato samples with 10 U/mL of PyASNase at 85 °C for merely 10 min reduced the acrylamide content in the final product by 82.5%, demonstrating a high efficiency and significant advantage of PyASNase in acrylamide inhibition.

Hydrometallurgical bioprocesses for base metal recovery in environmentally friendly electronic device waste (e-waste) recycling are typically studied under neutral pH conditions to avoid competition between metals and hydrogen ions. However, metal leachate is generally strongly acidic, thus necessitating a neutralisation process in the application of these bioprocesses to e-waste recycling. To solve this pH disparity, we focused on acid-tolerant bacteria for metal recovery under strongly acidic conditions. Four acid-tolerant bacterial strains were isolated from neutral pH environments to recover base metals from simulated waste metal leachate (pH 1.5, containing 100 or 1000 mg L^-1 of Co, Cu, Li, Mn, and Ni) without neutralisation. The laboratory setting for sequential metal recovery was established using these strains and a reported metal-adsorbing bacterium, Micrococcus luteus JCM1464. The metal species were successfully recovered from 100 mg L^-1 metal mixtures at the following rates: Co (8.95%), Cu (21.23%), Li (5.49%), Mn (13.18%), and Ni (9.91%). From 1000 mg L^-1 metal mixtures, Co (7.23%), Cu (6.82%), Li (5.85%), Mn (7.64%), and Ni (7.52%) were recovered. These results indicated the amenability of acid-tolerant bacteria to environmentally friendly base metal recycling, contributing to the development of novel industrial application of the beneficial but unutilised bioresource comprising acid-tolerant bacteria.

We acquired and analyzed metagenome and 16S/18S rRNA gene amplicon data of green-colored microbial mats from two hot springs within the Onikobe geothermal region (Miyagi Prefecture, Japan). The two collection sites-Tamago and Warabi-were in proximity and had the same temperature (40 °C), but the Tamago site was connected to a nearby stream, whereas the Warabi site was isolated. Both the amplicon and metagenome data suggest the bacterial, especially cyanobacterial, dominance of the mats; other abundant groups include Chloroflexota, Pseudomonadota, Bacteroidota/Chlorobiota, and Deinococcota. At finer resolution, however, the taxonomic composition entirely differed between the mats. A total of 5 and 21 abundant bacterial 16S rRNA gene OTUs were identified for Tamago and Warabi, respectively; of these, 12 are putative chlorophyll- or rhodopsin-based phototrophs. The presence of phylogenetically diverse microbial eukaryotes was noted, with ciliates and amoebozoans being the most abundant eukaryote groups for Tamago and Warabi, respectively. Fifteen metagenome-assembled genomes (MAGs) were obtained, represented by 13 bacteria, one ciliate (mitochondrion), and one giant virus. A total of 15 novel taxa, including a new deeply branching Chlorobiota species, is noted from the amplicon and MAG data, highlighting the importance of environmental sequencing in uncovering hidden microorganisms.

Methanogenic archaea are chemolithotrophic prokaryotes that can reduce carbon dioxide with hydrogen gas to form methane. These microorganisms make a significant contribution to the global carbon cycle, with methanogenic archaea from anoxic environments estimated to contribute > 500 million tons of global methane annually. Archaeal methanogenesis is dependent on the methanofurans; aminomethylfuran containing coenzymes that act as the primary C₁ acceptor molecule during carbon dioxide fixation. Although the biosynthetic pathway to the methanofurans has been elucidated, structural adaptations which confer thermotolerance to Mfn enzymes from extremophilic archaea are yet to be investigated. Here we focus on the methanofuran biosynthetic enzyme MfnB, which catalyses the condensation of two molecules of glyceralde-3-phosphate to form 4‑(hydroxymethyl)-2-furancarboxaldehyde-phosphate. In this study, MfnB enzymes from the hyperthermophile Methanocaldococcus jannaschii and the mesophile Methanococcus maripaludis have been recombinantly overexpressed and purified to homogeneity. Thermal unfolding studies, together with steady-state kinetic assays, demonstrate thermoadaptation in the M. jannaschii enzyme. Molecular dynamics simulations have been used to provide a structural explanation for the observed properties. These reveal a greater number of side chain interactions in the M. jannaschii enzyme, which may confer protection from heating effects by enforcing spatial residue constraints.