There are many species of Chinese traditional leguminosae family plants that are well known for their medicinal applications, such as Astragalus membranaceus, Catsia tora, Glycyrrhiza uralensis, Sophora flavescens and Albacia acacia. Their unique bioactive composition and internal phenological environment contribute to the formation of specific and unique endophytic fungal communities, which are important resources for new compounds used in a variety of pharmacological activities. Nonetheless, they have not been systematically studied. In the last decade, nearly 64 genera and thousands of species of endophytic fungi have been discovered from leguminosae plants, as well as 138 secondary metabolites (with 34 new compounds) including flavonoid, alkaloids, phenol, anthraquinone, macrolide, terpenoid, phytohormone and many more. These were shown to have diverse applications and benefits, such as antibacterial, antitumor, antioxidative, immunoregulatory and neuroprotective properties. Here, we provide a summarized overview with the aim of raising awareness of endophytic fungi from medicinal leguminosae plants and providing a comprehensive review of the discoveries of new natural products that may be of medicinal and pharmaceutical importance.

In this study, the hydroxy fatty acid dehydrogenase CLA-DH from Lactobacillus plantarump-8 and its four mutant variants were expressed in Escherichia coli Rosetta (DE3). UV spectrophotometry was employed to verify the catalytic power of the purified CLA-DH to convert ricinoleic acid into 12-oxo-cis-9-octadecenoic acid in the presence of oxidized nicotinamide adenine dinucleotide (NAD+). The optimum reaction temperature for CLA-DH was 45°C, with a maintained stability between 20°C and 40°C. The optimal pH for CLA-DH catalytic activity was 6.0-7.0, with a maintained stability at a pH range of 6.0-8.0. In addition, Fe3+ promoted enzyme activity, whereas Cu2+, Zn2+, and Fe2+ inhibited enzyme activity (P < 0.05). The Km, Vmax, Kcat, and Kcat/Km of CLA-DH were determined as 2.19 ± 0.34 μM, 2.06 ± 0.28 μM min-1, 2.00 ± 0.27 min-1, and 0.92 ± 0.02 min-1μM-1, respectively. Site-directed mutagenesis and molecular dynamics simulations demonstrated that both Tyr156 and Ser143 residues play significant roles in the catalysis of CLA-DH, and its solubility is affected by Lys160 and Asp63. Moreover, Gas chromatography determined that recombinant CLA-DH could be successfully applied to Conjugated linoleic acids production.

Biogenic coalbed methane is produced by biological processes mediated by synergistic interactions of microbial complexes in coal seams. However, the ecological role of functional bacteria in biogenic coalbed methane remains poorly understood. Here, we studied the metagenome assembled genomes (MAGs) of Bacillales and Clostridiales from coal seams, revealing further expansion of hydrogen and acetogen producers involved in organic matter decomposition. In this study, Bacillales and Clostridiales were dominant orders (91.85 ± 0.94%) in cultured coal seams, and a total of 16 MAGs from six families, including Bacillus, Paenibacillus, Staphylococcus, Anaerosalibacter, Hungatella and Paeniclostridium, were reconstructed. These microbial groups possessed multiple metabolic pathways (glycolysis/gluconeogenesis, pentose phosphate, β-oxidation, TCA cycle, assimilatory sulfate reduction, nitrogen metabolism and encoding hydrogenase) that provided metabolic substrates (acetate and/or H2) for the methanogenic processes. Therein, the hydrogenase-encoding gene and hydrogenase maturation factors were merely found in all the Clostridiales MAGs. β-oxidation was the main metabolic pathway involved in short-chain fatty acid degradation and acetate production, and most of these pathways were detected and exhibited different operon structures in Bacillales MAGs. In addition, assimilatory sulfate reduction and nitrogen metabolism processes were also detected in some MAGs, and these processes were also closely related to acetate production and/or organic matter degradation according to their operon structures and metabolic pathways. In summary, this study enabled a better understanding of the ecological roles of Bacillales and Clostridiales in biogenic methane in coal seams based on a combination of bioinformatic techniques.

Astronauts undergo space travel to bring scientific information to benefit humanity under various missions of space agencies such as NASA, European Space Agency, Indian Space Research Organization etc. During space missions, they encounter several stressors namely microgravity, fluid shifts, cosmic radiation, sleep deprivation and alteration in the circadian rhythm perturbing the quality of sleep. In addition, confined spaces makes pathogen interaction more likely if a pathobiont gets introduced into spacecraft. Microbiota is the first line оf resistаnсe tо vаriоus disorders and diseаses. It direсtly influenсes the biосhemiсаl, рhysiоlоgiсаl, аnd immunоlоgiсаl раthwаys. 'Gut microbiota' is essential for maintenance of healthy gut barrier functions. 'Dysbiosis' refers to perturbation of microbiota which is correlated with several metabolic and psychological disorders. Microbial metabolites are implicated in maintenance of human health. Investigations conducted on astronauts in international space missions and on analog terrestrial models have indicated a 'dysbiosis' of the gut microbiota associated with spaceflights. 'Dysbiosis' of the gut microbiome observed in astronauts has been implicated in immune dysregulation and a probiotic enriched diet is proposed to restore immune homeostasis. This article not just summarizes the state of art research on dysbiosis of the gut microbiome of astronauts, but also a diet mediated correction plan to restore their health especially during long term space missions. A characterization of microbial metabolites of the gut to enable administration of astronaut specific probiotic, postbiotic or synbiotic to alleviate space associated dysbiosis is proposed. It is also recommended that astronauts maintain a balanced nutritious diet throughout life to promote a resilient microbiota that is not perturbed by space missions. Further, a bioregenerative life support system wherein a probiotic may be produced in space station is proposed.

Patients with Crohn's disease (CD) have higher incidences of oral diseases such as dental caries and periodontitis than healthy people. Studies indicate that the interaction between gut and oral microbiota is an important factor. To compare the composition and diversity of the oral microbiome in periodontitis and CD-associated periodontitis, subgingival plaque and saliva samples from patients with these diseases were collected for 16S rRNA gene sequencing analyses. In CD-associated periodontitis, the subgingival plaque had greater microbial diversity than saliva. Subgingival plaque had decreased abundances of Firmicutes, Streptococcus, and Haemophilus and increased abundances of Bacteroidetes, Actinomyces, Treponema_2, Capnocytophaga, and Porphyromonas relative to saliva. The microbial composition in subgingival plaque was similar between the two diseases. Both red complex (Porphyromonas, Tannerella, and Treponema) and orange complex (Fusobacteria) bacteria were abundant in periodontitis subgingival plaque, while orange complex bacteria (Prevotella_2 and Prevotella) were abundant in CD-associated periodontitis subgingival plaque. Pocket depth was significantly positively correlated with multiple periodontal pathogens, including Porphyromonas, Tannerella, and Treponema. This study reveals the similarities and differences in the oral microbiome between periodontitis and CD-associated periodontitis, which provides a foundation to further explore the associations between CD and periodontitis.

Co-exposure to aflatoxin and fumonisin is a health concern where corn is a staple food, and a method to prevent co-contamination of these mycotoxins in foods is urgently needed. Polyoxins are chitin synthase inhibitors produced by Streptomyces cacaoi var. asoensis. The aflatoxin production inhibitory activity of a commercially available polyoxin D and four polyoxins purified from polyoxin AL water-soluble powder, an agricultural chemical containing polyoxins, was tested. The five polyoxins dose-dependently inhibited aflatoxin production by Aspergillus parasiticus and the IC50 values of polyoxin A, B, D, K and L were 16, 74, 110, 9 and 280 µmol L-1, respectively. Polyoxins also inhibited fumonisin production by Fusarium fujikuroi, and the IC50 values of polyoxin B, D, K and L were 270, 42, 65 and 62 µmol L-1, respectively. Polyoxins repressed the transcription of genes encoding proteins required for aflatoxin biosynthesis in A. parasiticus and fumonisin biosynthesis in F. fujikuroi. Polyoxin K and D also inhibited conidiation in A. parasiticus and F. fujikuroi, respectively. These results suggest that a mixture of polyoxins may effectively prevent co-contamination of aflatoxin and fumonisin in foods.

Prevalence of seafood-borne gastroenteritis caused by the human pathogen Vibrio parahaemolyticus is increasing globally despite current preventative measures. The United States Centers for Disease Control have designated V. parahaemolyticus as a reportable emerging human pathogen. The Eastern oyster (Crassostrea virginica) is a natural reservoir of the bacterium in marine environments, but little is actually known regarding interactions between oysters and V. parahaemolyticus. Therefore, a laboratory-scale Biosafety Level-2 (BSL2) inoculation system was developed wherein Chesapeake Bay region oysters harvested during summer or winter months, were exposed to the clinical RIMD2210633 strain carrying a chloramphenicol-selective marker (VP RIMDmC). Homogenized whole oyster tissues were spread on selective and differential agar medium to measure viable VP RIMDmC levels. Endogenous Vibrio spp. cell numbers were significantly reduced followed chloramphenicol treatment and this likely contributed to higher VP RIMDmC oyster-associated levels, especially using winter-harvested animals. Summer-harvested oysters had significantly higher existing Vibrio levels and a lower level of artificial oyster-associated VP RIMDmC. Thus, the pre-existing microbiome appears to afford some protection from an external V. parahaemolyticus challenge. Overall, this system successfully enabled controlled manipulation of parameters influencing V. parahaemolyticus-oyster interactions and will be useful in safely testing additional pertinent environmental variables and potential mitigation strategies.

Substrate uptake and product export are important for microbial growth and product synthesis. Here, the glycerol uptake facilitator (GlpF) and the members of the resistance-nodulation-cell division (RND) type efflux system were overexpressed in Klebsiella pneumoniae to promote 1,3-propanediol (1,3-PDO) production. Overexpression of the endogenous K. pneumoniae GlpF improved glycerol dehydratase (GDHt) activity and promoted 1,3-PDO titer from 55.6 to 65.1 g/l. RND members AcrA and the AcrE had no impact on 1,3-PDO production. RND members AcrF and the TolC increased 1,3-PDO titer from 55.6 to 68.4 g/l and 65.4 g/l, respectively. MexB significantly decreased GDHt activity and 1,3-PDO titer. Notably, MexF dramatically enhanced GDHt activity and promoted 1,3-PDO titer and glycerol conversion rate to 74.0 g/l and 0.62 mol/mol, respectively. However, coexpression of the endogenous GlpF and MexF did not further improve 1,3-PDO production. The results present here provided novel information about the applications of the uptake of glycerol and the efflux of 1,3-PDO.

The cost of microalgae cultivation is one of the largest limitations to achieving sustainable, large-scale microalgae production of commercially desirable lipids. Utilizing CO2 as a 'free' carbon source from waste industrial flue gas emissions can offer wide-ranging cost savings. However, these gas streams typically create acidic environments, in which most microalgae cannot survive due to the concentration of CO2 and the presence of other acidic gasses such as NO2 and SO2. To address this situation, we investigated growth of a mixed acid-tolerant green microalgal culture (91% dominated by a single Coccomyxa sp. taxon) bioprospected at pH 2.8 from an acid mine drainage impacted water body. The culture was grown at pH 2.5 and fed with a simulated flue gas containing 6% CO2 and 94% N2. On reaching the end of the exponential growth phase, the culture was exposed to either continued light-dark cycle conditions or continual dark conditions. After three days in the dark, the biomass consisted of 28% of lipids, which was 42% higher than at the end of the exponential phase and 55% higher than the maximum lipid content achieved under light/dark conditions. The stress caused by being continually in the dark also favoured the production of omega-3 and omega-6 polyunsaturated fatty acids (PUFAs; 19.47% and 21.04%, respectively, after 7 days) compared to 7-days of light-dark treatment (1.94% and 9.53%, respectively) and showed an increase in nitrogen content (C:N ratio of 6.4) compared to light-dark treatment (C:N ratio of 11.9). The results of the research indicate that use of acid tolerant microalgae overcomes issues using flue gasses that will create an acidic environment and that applying dark stress is a low-cost stressor stimulates production of desirable dietary lipids.

Molecular biology theory represents a critical scaffold, which underpins multiple disciplines within life sciences education. However, it is well-documented that undergraduate students can struggle to achieve deeper understanding of key concepts and/or their application. One challenging, contributory aspect is the "invisible" nature of molecular biology processes compounded by critical 3D spatial orientations of the principal components and their interactions. Molecular theory specifically requires students to construct accurate, mental spatial models to develop their understanding. However, much of the traditional teaching and examination of such theory is limited to 2D representations. Technology-enhanced, complementary teaching and examination approaches, which engage students with spatial aspects of theoretical concepts, offer an exciting opportunity to support student learning in this area. In this study, we have explored the integration of an immersive virtual reality simulation based on a challenging molecular biology concept within an existing module taught at University College Cork. A mixed methods approach, grounded in learning theory, was undertaken to assess the student user and learning experience. The consensus response from students was one of enhanced learning, understanding, engagement, and motivation. Student partnership in the process of simulation design and integration was key to delivering the fully integrated experience.

Outer-membrane vesicles (OMVs) are promising tools in the development of novel vaccines against the respiratory pathogens Bordetella pertussis and Bordetella bronchiseptica. Unfortunately, vesiculation by bordetellae is too low for cost-effective vaccine production. In other bacteria, iron limitation or inactivation of the fur gene has been shown to increase OMV production, presumably by downregulation of the mla genes, which encode machinery for maintenance of lipid asymmetry in the outer membrane. Here, we followed a similar approach in bordetellae. Whereas a fur mutant was readily obtained in B. bronchiseptica, a B. pertussis fur mutant could only be obtained in iron-deplete conditions, indicating that a fur mutation is conditionally lethal in this bacterium. The fur mutants displayed a growth defect in iron-replete media, presumably because constitutive expression of iron-uptake systems resulted in iron intoxication. Accordingly, expression of the Escherichia coli ferritin FtnA to sequester intracellularly accumulated iron rescued the growth of the mutants in these media. The fur mutations led to the constitutive expression of novel vaccine candidates, such as the TonB-dependent receptors FauA for the siderophore alcaligin and BhuR for heme. However, neither inactivation of fur nor growth under iron limitation improved vesiculation, presumably because the expression of the mla genes appeared unaffected.

A moderate halophilic bacterium that could accumulate ectoine and hydroxyectoine was isolated from soil near a salt mine and was identified as a Sinobaca sp. (designed strain H24) according to 16S rRNA gene sequence analysis. The bacterium grew well in the presence of 1-2 M NaCl, while growth in a medium that contained 2 M NaCl led to higher accumulation of ectoines. The yields of ectoine and hydroxyectoine by Sinobaca sp. H24 reached 11.27 mg/l and 1.34 mg/l, respectively, when cultured in the following medium: NaCl (2 M), peptone (5 g/l), yeast extract (1 g/l), NH4Cl (0.02 M), KH2PO4 (1 M), K2HPO4 (0.1 M), and glycerol (1% w/v). Genes that are involved in ectoine biosynthesis of Sinobaca sp. H24 were also identified, and their sequences were determined by a metagenomics approach. The results demonstrated that Sinobaca sp. H24 possesses ectoine metabolism genes for both ectoine biosynthesis (ectA, ectB, ectC, and ectD) and ectoine degradation (doeA). Genes that are related to ectoine biosynthesis, such as lysC and asd, were also characterized. The identification and characterization results for ectoine/hydroxyectoine biosynthesis genes are in agreement with the physiology of Sinobaca sp. H24 as a potential candidate for ectoine production for industrial applications. This report established for the first time the accumulation of ectoine/hydroxyectoine in Sinobaca sp. and characterized the genes that are involved in ectoine/hydroxyectoine biosynthesis in Sinobaca sp. H24.

Blue light (BL) has shown bactericidal effectiveness against methicillin-resistant Staphylococcus aureus (MRSA), one of the major clinical pathogens with antibiotic resistance. Bacteria likely respond to the oxidative stress induced by BL; however, the defensive response is still unclear. This study aimed to reveal the phenotypic change in MRSA after being exposed to 15 cycles of sub-lethal BL illumination. The comparative transcriptomic results showed that the expression of peptidoglycan (PG) synthesis gene glmS was significantly upregulated in the cells after the multiple cycle light treatment, and the biochemical analysis determined that the content of PG synthesized was increased by 25.86% when compared with that in control cells. Furthermore, significant thickening of the cell wall was observed under a transmission electron microscope (P < .05). The light sensitivity of the tested MRSA strain was reduced after the multiple cycle light treatment, indicating the possibility of MRSA being more adaptive to the BL stress. The present study suggested that multiple cycles of sub-lethal BL could change the light susceptibility of MRSA through thickening the cell wall.

The diversity of fenugreek (Trigonella foenum-graecum L.) microsymbionts has been barely studied even though it is of great interest for being a spice and a medicinal plant. Here, we analyzed 59 bacterial strains isolated from fenugreek nodules originating from different geographic and climatic areas of Iran. Most of these strains exhibit phenotypic characteristics compatible with rhizobia and they nodulate fenugreek. Analysis of the recA and atpD genes shows that representative strains of ERIC-BOX-PCR groups cluster with the type strains of Ensifer meliloti and E. kummerowiae as well as with strains capable of nodulating different Trigonella species found in other countries. The closeness of E. meliloti and E. kummerowiae suggests there is a need to revise the taxonomic status of the latter species. The nodC gene analysis shows that most Trigonella-nodulating strains belong to the symbiovar meliloti except those nodulating Trigonella arcuata in China, which belong to the symbiovar rigiduloides. This analysis shows that the type strains of E. kummerowiae, E. meliloti, and E. medicae belonged to three well-defined groups within the symbiovar meliloti, with the Iranian strains belonging to the E. kummerowiae subgroup. The small group of strains unable to nodulate fenugreek isolated in this study belong to Enterobacter cloacae, reported for the first time as being a possible endophyte of fenugreek nodules.