Exopolysaccharide Biosynthesis Research Articles

Valorization of biodiesel-derived crude glycerol for simultaneous biosynthesis of biodegradable polyhydroxybutyrate and exopolysaccharide by the newly isolated Burkholderia sp. SCN-KJ

This study demonstrated that Burkholderia sp. SCN-KJ is a promising novel species for the biovalorization of crude glycerol to polyhydroxybutyrate (PHB) and galactose-rich heteroexopolysaccharide (EPS). Whole-genome and genetic evolution analyses revealed separation of the different clades according to the ANIb and dDDH analyses, which confirmed that Burkholderia sp. SCN-KJ is a novel species. The highest PHB production from crude glycerol was 12.9 ± 0.4 g/L (72.9 ± 2.1 % w/w), with a productivity of 0.46 g/L/h and YP/S of 0.3 g/g at 28 h in a 10 L fermenter. The galactose-rich hetero-EPS began to be produced after nitrogen depletion, resulting in a concentration of 22.4 ± 0.2 g/L at 38 h. Examination of the carbon-to‑nitrogen ratio (C/N) showed that nitrogen-rich condition (C/N 20) was optimal for PHB production, whereas nitrogen-depleted condition promoted EPS production, showing two different extrema. The findings showed that Burkholderia sp. SCN-KJ has the potential to transform the landscape of biovalorization for sustainable production.

Understanding Bacillus response to salt stress: Growth inhibition, enhanced EPS secretion, and molecular adaptation mechanisms

This study investigates the secretion pattern of extracellular polymeric substances (EPS) by Bacillus sp. under varying salt concentrations and elucidates the molecular mechanisms governing EPS synthesis and secretion. Salt stress inhibited cell proliferation, while optimal salt stimulation promoted EPS secretion, resulting in increased viscosity of the culture medium and the formation of bacterial clusters. Fourier infrared spectrum analysis revealed functional groups such as C-O-C and N-H within the EPS. Soluble-EPS (S-EPS) contained sulfur and phosphorus groups associated with heavy metal ions adsorption. The study also identified a novel polysaccharide formed through bonding EPS (B-EPS). High salt concentrations correlated with elevated levels of tryptophan protein and its derivatives, increased tyrosine polysaccharide derivatives, and decreased aromatic polysaccharides. B-EPS exhibited higher levels of aromatic polysaccharides, with Na+ promoting detachment of B-EPS from the cell surface. Transcriptome sequencing (RNA-seq) analysis under salt stress revealed significant expression of spore kinase (KinD) and response regulatory protein Spo0A in the phosphoric acid relay system. Key transcriptional regulatory factors, including OmpR and exopolysaccharide biosynthesis, were closely associated with EPS synthesis and secretion. This study establishes a theoretical foundation for the industrial production and practical application of EPS by elucidating the molecular mechanisms underlying Bacillus' response to salt stress.

Characterization of a novel antioxidant exopolysaccharide from an intestinal-originated bacteria Bifidobacterium pseudocatenulatum Bi-OTA128

Microbial exopolysaccharides (EPSs) have attracted extensive attention for their biological functions in antioxidant activities. In this study, we characterized a novel EPS produced by Bifidobacterium pseudocatenulatum Bi-OTA128 which exhibited the highest antioxidant capacity compared to nine other ropy bacterial strains, achieving 76.50 % and 93.84 % in DPPH· and ABTS·+ scavenging activity, and ferric reducing power of 134.34 μM Fe2+. Complete genomic analysis identified an eps gene cluster involved in the EPS biosynthesis of Bi-OTA128 strain, which might be responsible for its ropy phenotype. The EPS was then isolated and purified by a DEAE-Sepharose Fast Flow column. A single elution part EPS128 was obtained with a recovery rate of 43.5 ± 1.78 % and a total carbohydrate content of 93.6 ± 0.76 %. Structural characterization showed that EPS128 comprised glucose, galactose, and rhamnose (molar ratio 4.0:1.2:1.1), featuring a putative complex backbone structure with four branched chains and an unusual acetyl group at O-2 of terminal rhamnose. Antioxidant assay in vitro indicated that EPS128 exhibited antioxidant potential with 50.52 % DPPH· and 65.40 % ABTS·+ scavenging activities, reaching 54.3 % and 70.44 % of the efficacy of standard Vitamin C at 2.0 mg/L. Furthermore, EPS128 showed protective effects against H2O2-induced oxidative stress in HepG2 cells by reducing cellular reactive oxygen species (ROS) and increasing cell viability. These findings present the first comprehensive report of an antioxidant EPS from B. pseudocatenulatum, highlighting its potential as a natural antioxidant for applications in the food industry and clinical settings.

Exopolysaccharides as bio-based rheology modifiers from microalgae produced on dairy industry waste: Towards a circular bioeconomy approach

The feasibility of exopolysaccharides (EPS) production from cheese whey using Chlorella vulgaris was investigated as an example of circular bioeconomy application. The effects of dairy waste utilization in EPS biosynthesis and rheological properties were evaluated, comparing with both control conditions and commercial xanthan gum (CXG). A twofold increase in yield, up to 0.32 g L−1, was observed when Chlorella vulgaris was used for EPS production from whey rather than conventional fertilizers. Additionally, the EPS produced using cheese whey exhibited superior pseudoplasticity in the 0.4–1.0 (w/v) range compared to the control. The EPS from the whey wastewater contained functional groups similar to those of CXG (82.7 %). Moreover, the solutions containing 1 % biopolymer showed rheological profiles similar to those of the 0.4 % CXG. The molecular weight averages predominantly fell within the range of 284 to 324 kDa, as deduced using diffusion NMR, an innovative and rapid determination method for estimating EPS size. The potential applications of EPS notably extend beyond the dairy industry, reaching diverse market sectors, and thereby enhancing the competitiveness of microalgal biorefineries while contributing to the achievement of Sustainable Development Goals.

Mechanism and application of bacterial exopolysaccharides: An advanced approach for sustainable heavy metal abolition from soil

The escalation of heavy metal pollutants in soils and effluents, driven by industrialization and human activities, poses significant environmental and health risks. Conventional remediation methods are often costly and ineffective, prompting a shift towards sustainable alternatives such as biological treatments. Natural biosorbents, including microbial cells and their byproducts, have emerged as promising solutions. One such approach involves leveraging exopolysaccharides (EPS), complex high-molecular-weight biopolymers synthesized by microbes under environmental stress conditions. EPS are intricate organic macromolecules comprising proteins, polysaccharides, uronic acids, humic compounds, and lipids, either located within microbial cells or secreted into their surroundings. Their anionic functional groups enable efficient electrostatic binding of cationic heavy metals, making EPS effective biosorbents for soil remediation. This review thoroughly explores the pivotal role of bacterial EPS in the removal of heavy metals, focusing on EPS biosynthesis mechanisms, the dynamics of interaction with heavy metals, and case studies that illustrate their effectiveness in practical remediation strategies. By highlighting these aspects, the review underscores the innovation and practical implications of EPS-based bioremediation technologies, demonstrating their potential to address critical environmental challenges effectively while paving the way for sustainable environmental management practices. Key findings reveal that EPS exhibit robust metal-binding capacities, facilitated by their anionic functional groups, thereby offering a promising solution for mitigating metal pollution in diverse environmental matrices.

Bioproduction of exopolysaccharides by lactic acid bacteria using agave by-products

The capacity to produce exopolysaccharides (EPS) is well known among several microorganisms, especially lactic acid bacteria (LAB). However, the physiological role of these polymers has yet to be elucidated. Nevertheless, it has been observed that LAB growth conditions and adaptation to external environments and substrates are directly related to EPS biosynthesis. This study aimed to evaluate alternative carbon sources from agave by-products to synthesize EPS with LAB. Native LAB from the genera Lactococcus and Lactiplantibacillus were evaluated for their ability to produce EPS from simple carbon sources; they showed potential to produce EPS, presenting positive results on Congo red plates using glucose and fructose as carbon sources. In addition, native LAB could synthesize EPS from agave leaves and bagasse. L. plantarum ITD-ZC-107 stand out in the production of EPS, especially from bagasse. Thermostability of EPS from the evaluated carbon sources was analyzed. EPS from agave leaves and bagasse showed a higher degradation temperature than the ones obtained from glucose and sucrose, indicating better thermo-resistance.

Optimization of the production of Exopolysaccharide (EPS) from biofilm-forming bacterial consortium using different parameters

Bacterial cells produce a variety of exopolysaccharide (EPS) throughout their life cycle. EPS plays a crucial role in the formation of biofilms and the control of environmental processes in microbial communities. The bacterial community benefits from biofilms in several ways, including nutrient compartmentation and storage, environmental stress resistance and synergism. The generation of EPS by consortium-associated bacteria, which involves a variety of microorganisms, is versatile and efficient for use in industrial and environmental processes, such as biotechnology and pollution degradation. The qualitative and quantitative chemical characteristics of the consortium's EPS are mostly determined by the type of culture utilized, nutrient type, temperature, and pH. The main goal of the study was to find out how EPS biosynthesis works in a bacterial consortium culture. To achieve maximum EPS regeneration, pH, temperature, carbon, and nitrogen sources were optimized. A consortium of three bacterial strains i.e. Klebsiella pneumonia strain, Pseudomonas aeruginosa strain and Burkholderia cepacian strain that were distinguished by their capacity to produce biofilms were developed as part of the study. The maximum total EPS production was detected after 8 days of incubation at pH 7.5 (15.5 g/L) and a temperature of 35 °C (12.60 g/L). The optimum EPS production was identified as 17.84 g/L and 21.07 g/L when ammonium sulfate and glucose were employed as nitrogen and carbon supplements, respectively. The protein concentration of the EPS was 3067 µg/mL, which was more than the carbohydrate (263.10 µg/mL) and DNA (7.60 µg/mL) content. The total protein/carbohydrate ratio of the EPS detected in the present study was 11.65, which was much higher than in prior investigations. FTIR analysis of EPS confirmed the functional groups of carbohydrates, proteins, lipids and DNA, viz., CO, COOH, NH, OH, CH and OCH3. The study's findings revealed that enhancing the consortium's EPS molecules involves lowering biofilm-related infections and improving the efficacy of antimicrobial treatments in wastewater.

Transcriptome Analysis Reveals the Role of Sucrose in the Production of Latilactobacillus sakei L3 Exopolysaccharide.

Latilactobacillus (L.) sakei is a species of lactic acid bacteria (LAB) mostly studied according to its application in food fermentation. Previously, L. sakei L3 was isolated by our laboratory and possessed the capability of high exopolysaccharide (EPS) yield during sucrose-added fermentation. However, the understanding of sucrose promoting EPS production is still limited. Here, we analyzed the growth characteristics of L. sakei L3 and alterations of its transcriptional profiles during sucrose-added fermentation. The results showed that L. sakei L3 could survive between pH 4.0 and pH 9.0, tolerant to NaCl (<10%, w/v) and urea (<6%, w/v). Meanwhile, transcriptomic analysis showed that a total of 426 differentially expressed genes and eight non-coding RNAs were identified. Genes associated with sucrose metabolism were significantly induced, so L. sakei L3 increased the utilization of sucrose to produce EPS, while genes related to uridine monophosphate (UMP), fatty acids and folate synthetic pathways were significantly inhibited, indicating that L. sakei L3 decreased self-growth, substance and energy metabolism to satisfy EPS production. Overall, transcriptome analysis provided valuable insights into the mechanisms by which L. sakei L3 utilizes sucrose for EPS biosynthesis. The study provided a theoretical foundation for the further application of functional EPS in the food industry.

Glycosyltransferase Slr1064 regulates carbon metabolism by modulating the levels of UDP-GlcNAc in Synechocystis sp. PCC 6803.

Glycosyltransferases (GTs) are enzymes that transfer sugars to various targets. They play important roles in diverse biological processes, including photosynthesis, cell motility, exopolysaccharide biosynthesis, and lipid metabolism; however, their involvement in regulating carbon metabolism in Synechocystis sp. PCC 6803 has not been reported. We identified a novel GT protein, Slr1064, involved in carbon metabolism. The effect of slr1064 deletion on the growth of Synechocystis cells and functional mechanisms of Slr1064 on carbon metabolism were thoroughly investigated through physiological, biochemistry, proteomic, and metabolic analyses. We found that this GT, which is mainly distributed in the membrane compartment, is essential for the growth of Synechocystis under heterotrophic and mixotrophic conditions, but not under autotrophic conditions. The deletion of slr1064 hampers the turnover rate of Gap2 under mixotrophic conditions and disrupts the assembly of the PRK/GAPDH/CP12 complex under dark culture conditions. Additionally, UDP-GlcNAc, the pivotal metabolite responsible for the O-GlcNAc modification of GAPDH, is downregulated in the Δslr1064. Our work provides new insights into the role of GTs in carbon metabolism in Synechocystis and elucidate the mechanism by which carbon metabolism is regulated in this important model organism.

Genomic and functional evaluation of exopolysaccharide produced by Liquorilactobacillus mali t6-52: technological implications

BackgroundThis study explores the biosynthesis, characteristics, and functional properties of exopolysaccharide produced by the strain Liquorilactobacillus mali T6-52. The strain demonstrated significant EPS production with a non-ropy phenotype.ResultsThe genomic analysis unveiled genes associated with EPS biosynthesis, shedding light on the mechanism behind EPS production. These genes suggest a robust EPS production mechanism, providing insights into the strain’s adaptability and ecological niche. Chemical composition analysis identified the EPS as a homopolysaccharide primarily composed of glucose, confirming its dextran nature. Furthermore, it demonstrated notable functional properties, including antioxidant activity, fat absorption capacity, and emulsifying activity. Moreover, the EPS displayed promising cryoprotective activities, showing notable performance comparable to standard cryoprotective agents. The EPS concentration also demonstrated significant freeze-drying protective effects, presenting it as a potential alternative cryoprotectant for bacterial storage.ConclusionsThe functional properties of L. mali T6-52 EPS reveal promising opportunities across various industrial domains. The strain’s safety profile, antioxidant prowess, and exceptional cryoprotective and freeze-drying characteristics position it as an asset in food processing and pharmaceuticals.

New insight into protective effect against oxidative stress and biosynthesis of exopolysaccharides produced by Lacticaseibacillus paracasei NC4 from fermented eggplant.

Fermented eggplant is a traditional fermented food, however lactic acid bacteria capable of producing exopolysaccharide (EPS) have not yet been exploited. The present study focused on the production and protective effects against oxidative stress of an EPS produced by Lacticaseibacillus paracasei NC4 (NC4-EPS), in addition to deciphering its genomic features and EPS biosynthesis pathway. Among 54 isolates tested, strain NC4 showed the highest EPS yield and antioxidant activity. The maximum EPS production (2.04 ± 0.11g/L) was achieved by culturing in MRS medium containing 60g/L sucrose at 37°C for 48h. Under 2mM H2O2 stress, the survival of a yeast model Saccharomyces cerevisiae treated with 0.4mg/mL NC4-EPS was 2.4-fold better than non-treated cells, which was in agreement with the catalase and superoxide dismutase activities measured from cell lysates. The complete genome of NC4 composed of a circular chromosome of 2,888,896bp and 3 circular plasmids. The NC4 genome comprises more genes with annotated function in nitrogen metabolism, phosphorus metabolism, cell division and cell cycle, and iron acquisition and metabolism as compared to other reported L. paracasei. Of note, the eps gene cluster is not conserved across L. paracasei. Pathways of sugar metabolism for EPS biosynthesis were proposed for the first time, in which gdp pathway only present in few plant-derived bacteria was identified. These findings shed new light on the cell-protective activity and biosynthesis of EPS produced by L. paracasei, paving the way for future efforts to enhance yield and tailor-made EPS production for food and pharmaceutical industries.

Investigation on the exopolysaccharide production from blueberry juice fermented with lactic acid bacteria: Optimization, fermentation characteristics and Vis-NIR spectral model

The exopolysaccharide production from blueberry juice fermented were investigated. The highest exopolysaccharide yield of 2.2 ± 0.1 g/L (increase by 32.5 %) was reached under the conditions of temperature 26.5 °C, pH 5.5, inoculated quantity 5.4 %, and glucose addition 9.1 % using the artificial neural network and genetic algorithm. Under the optimal conditions, the viable cell counts and total acids were increased by 2.0 log CFU/mL and 1.6 times, respectively, while the content of phenolics and anthocyanin was decreased by 9.26 % and 7.86 %, respectively. The changes of these components affected the exopolysaccharide biosynthesis. The absorption bands of –OH and –CH associated with the main functional groups of exopolysaccharide were detected by Visible near-infrared spectroscopy. The prediction model based on spectrum results was constructed. Competitive adaptive reweighted sampling and the random forest were used to enhance the model's prediction performance with the value of RC = 0.936 and RP = 0.835, indicating a good predictability of exopolysaccharides content during fermentation.

Characterization and functional evidence for Orf2 of Streptomyces sp. 139 as a novel dipeptidase E

Aspartyl dipeptidase (dipeptidase E) can hydrolyze Asp-X dipeptides (where X is any amino acid), and the enzyme plays a key role in the degradation of peptides as nutrient sources. Dipeptidase E remains uncharacterized in Streptomyces. Orf2 from Streptomyces sp. 139 is located in the exopolysaccharide biosynthesis gene cluster, which may be a novel dipeptidase E with “S134-H170-D198” catalytic triad by sequence and structure comparison. Herein, recombinant Orf2 was expressed in E. coli and characterized dipeptidase E activity using the Asp-ρNA substrate. The optimal pH and temperature for Orf2 are 7.5 and 40 ℃; Vmax and Km of Orf2 are 0.0787 mM·min−1 and 1.709 mM, respectively. Orf2 exhibits significant degradation activities to Asp-Gly-Gly, Asp-Leu, Asp-His, and isoAsp-Leu and minimal activities to Asp-Pro and Asp-Ala. Orf2 contains a Ser-His-Asp catalytic triad characterized by point mutation. In addition, the Asp147 residue of Orf2 is also proven to be critical for the enzyme’s activity through molecular docking and point mutation. Transcriptome analysis reveals the upregulation of genes associated with ribosomes, amino acid biosynthesis, and aminoacyl-tRNA biosynthesis in the orf2 mutant strain. Compared with the orf2 mutant strain and WT, the yield of crude polysaccharide does not change significantly. However, crude polysaccharides from the orf2 mutant strain exhibit a wider range of molecular weight distribution. The results indicate that the Orf2 links nutrient stress to secondary metabolism as a novel dipeptidase E.Key points• A novel dipeptidase E with a Ser-His-Asp catalytic triad was characterized from Streptomyces sp. 139.• Orf2 was involved in peptide metabolism both in vitro and in vivo.• Orf2 linked nutrient stress to mycelia formation and secondary metabolism in Streptomyces.

Physiological properties evaluation and exopolysaccharide biosynthesis of Limosilactobacillus fermentum JL-2 from Chinese traditional fermented dairy product based on genomic insight

Limosilactobacillus fermentum JL-2 with high production of exopolysaccharide (EPS) was screened from the traditional fermented dairy product in Xinjiang district (China) in our previous work. In this study, the whole genome of L. fermentum JL-2 was sequenced and the genomic information and function were studied to excavate its genotype, safety and probiotic properties. The safety evaluation in vitro showed that L. fermentum JL-2 was not hemolytic, sensitive to common antibiotics, and it had a weak biogenic amine production capacity. The genes related to EPS biosynthesis and their relationship with EPS production were discussed. The EPS production of L. fermentum JL-2 was 553.6 mg/L. The genome of L. fermentum JL-2 consisted of a circular 1.98 Mbp chromosome. A variety of genes related to probiotic properties such as acid tolerance, bile salt tolerance, adhesion to gut mucosa and environmental stress resistance were contained. A complete EPS gene cluster was found in the genome of L. fermentum JL-2, and the genes related to EPS biosynthesis such as epsA and epsB were found in L. fermentum JL-2. It is worth noting that EPS biosynthesis protein (gene0099) seemed to play a key role in the EPS synthesis of L. fermentum JL-2. These results revealed the potential of L. fermentum JL-2 as a probiotic at the gene level. Therefore, this study laid a theoretical foundation for further research into its potential probiotic function and its application in practice.

Effects of priming glycosyltransferase genes cps 2E and cps 4E on the exopolysaccharide biosynthesis by Lactiplantibacillus plantarum YM-4-3 strain

Microbial exopolysaccharides (EPS) have various physiological functions such as antioxidant, anti-tumor, cholesterol lowering, and immune regulation. However, improving traditional fermentation conditions to increase the production of EPS from Lactiplantibacillus plantarum (L. plantarum) is limited. In this study, we aimed to better improve EPS production and physiological functions of L. plantarum YM-4-3 strain by overexpressing and knocking out the priming glycosyltransferase genes cps 2E and cps 4E for the first time. As a result, the EPS production of the overexpression strain was 30.15 %, 26.84 % and 36.29 % higher than WT, respectively. The EPS production of the knockout strain was significantly lower than that of the WT. At the same time, transcriptome data showed that the gene expression levels of each experimental strain had changed. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways found that the glycolysis/gluconeogenesis pathway had the highest gene enrichment in the metabolic pathway. The monosaccharide components of the EPS of each experimental strain were different from those of the WT and the EPS of the experimental strain showed stronger activity against oxidation. In conclusion, this study contributes to the efficient production and application of L. plantarum EPS and helps to understand the mechanism of EPS regulation in L. plantarum.

Abstract 2801: Identifying gut microbial determinants of clonal hematopoiesis of indeterminate potential (CHIP) in immunotherapy treated melanoma patients

Abstract INTRO: Clonal hematopoiesis (CH) of indeterminate potential (CHIP) is associated with poor outcomes in those with non-hematologic cancers. CH mutated macrophages and neutrophils have increased capacity to home to peripheral tissues, driving local inflammation; they are also implicated in immunotherapy related toxicity. The relationship between CH and the microbiome in those treated with immunotherapy is not well understood. RESULTS: 35 patients with stage IV melanoma treated with upfront immunotherapy; treatment and cohort characteristics are below (Table 1). We performed error corrected duplex sequencing on peripheral blood mononuclear cells using a panel previously validated for CH mutations. With a cutoff of 0.5% variant allele frequency (VAF), among 10 CHIP positive patients, we found mutations in TET2 (8/10), DNMT3A (7/10), PPM1D (2/10), and JAK2 (1/10). There was no difference in either survival or response based on CHIP status (Log-Rank Chi-Square p=0.44); however, CHIP positivity was associated with greater microbiome richness as measured by alpha diversity (p=0.0012 Shannon diversity) and distinct structural/compositional diversity vs CHIP negative patients as measured by beta diversity (p=0.032, R Squared= 0.045). We inferred KEGG pathway activity from whole genome sequencing and identified that CHIP positive patients were enriched for exopolysaccharide biosynthesis while CHIP negative patients had higher expression of nucleotide sugars biosynthesis and amino acid metabolism pathways. DISCUSSION: In patients treated with immunotherapy for melanoma, microbiome diversity signatures correlate with presence of CHIP. The analysis suggests that there are distinct taxonomic and functional features defining CHIP positivity. The effect of these cells in the tumor microenvironment and their role in immunotherapy response requires further exploration. Citation Format: Eleanor A. Fallon, Samuel Urrutia, Reed Ayabe, Manoj Chelvanambi, Ashish Damania, Sarah Johnson, Tomoyuki Tanaka, Zongrui Li, Yongwoo David Seo, Samuel Cass, Matthew C. Wong, Nadim Ajami, Jennifer Wargo, Koichi Takahashi. Identifying gut microbial determinants of clonal hematopoiesis of indeterminate potential (CHIP) in immunotherapy treated melanoma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2801.

Analysis of the complete genome sequence of Paenibacillus sp. lzh-N1 reveals its antagonistic ability

BackgroundPlant diseases caused by pathogenic fungi are devastating. However, commonly used fungicides are harmful to the environment, and some are becoming ineffective due to fungal resistance. Therefore, eco-friendly biological methods to control pathogenic fungi are urgently needed.ResultsIn this study, a strain, Paenibacillus sp. lzh-N1, that could inhibit the growth of the pathogenic fungus Mycosphaerella sentina (Fr) Schrorter was isolated from the rhizosphere soil of pear trees, and the complete genome sequence of the strain was obtained, annotated, and analyzed to reveal the genetic foundation of its antagonistic ability. The entire genome of this strain contained a circular chromosome of 5,641,488 bp with a GC content of 45.50%. The results of species identification show that the strain belongs to the same species as P. polymyxa Sb3-1 and P. polymyxa CJX518. Sixteen secondary metabolic biosynthetic gene clusters were predicted by antiSMASH, including those of the antifungal peptides fusaricidin B and paenilarvins. In addition, biofilm formation-related genes containing two potential gene clusters for cyclic lactone autoinducer, a gene encoding S-ribosylhomocysteine lyase (LuxS), and three genes encoding exopolysaccharide biosynthesis protein were identified.ConclusionsAntifungal peptides and glucanase biosynthesized by Paenibacillus sp. lzh-N1 may be responsible for its antagonistic effect. Moreover, quorum sensing systems may influence the biocontrol activity of this strain directly or indirectly.

Metagenomic assembled genomes indicated the potential application of hypersaline microbiome for plant growth promotion and stress alleviation in salinized soils.

The strategic implementation of metagenomic assembled genomes (MAGs) in exploring the properties and harnessing microorganisms from ecosystems like hypersaline niches has transformative potential in agriculture. This approach promises to redefine our comprehension of microbial diversity and its ecosystem roles. Recovery and decoding of MAGs unlock genetic resources, enabling the development of new solutions for agricultural challenges. Enhanced understanding of these microbial communities can lead to more efficient nutrient cycling, pest control, and soil health maintenance. Consequently, traditional agricultural practices can be improved, resulting in increased yields, reduced environmental impacts, and heightened sustainability. MAGs offer a promising avenue for sustainable agriculture, bridging the gap between cutting-edge genomics and practical field applications.

Bacterial Endophytes from Legumes Native to Arid Environments Are Promising Tools to Improve Mesorhizobium-Chickpea Symbiosis under Salinity.

Symbiotic nitrogen fixation is a major contributor of N in agricultural ecosystems, but the establishment of legume-rhizobium symbiosis is highly affected by soil salinity. Our interest is focused on the use of non-rhizobial endophytes to assist the symbiosis between chickpea and its microsymbiont under salinity to avoid loss of production and fertility. Our aims were (1) to investigate the impact of salinity on both symbiotic partners; including on early events of the Mesorhizobium-chickpea symbiosis, and (2) to evaluate the potential of four non-rhizobial endophytes isolated from legumes native to arid regions (Phyllobacterium salinisoli, P. ifriqiyense, Xanthomonas translucens, and Cupriavidus respiraculi) to promote chickpea growth and nodulation under salinity. Our results show a significant reduction in chickpea seed germination rate and in the microsymbiont Mesorhizobium ciceri LMS-1 growth under different levels of salinity. The composition of phenolic compounds in chickpea root exudates significantly changed when the plants were subjected to salinity, which in turn affected the nod genes expression in LMS-1. Furthermore, the LMS-1 response to root exudate stimuli was suppressed by the presence of salinity (250 mM NaCl). On the contrary, a significant upregulation of exoY and otsA genes, which are involved in exopolysaccharide and trehalose biosynthesis, respectively, was registered in salt-stressed LMS-1 cells. In addition, chickpea co-inoculation with LMS-1 along with the consortium containing two non-rhizobial bacterial endophytes, P. salinisoli and X. translucens, resulted in significant improvement of the chickpea growth and the symbiotic performance of LMS-1 under salinity. These results indicate that this non-rhizobial endophytic consortium may be an appropriate ecological and safe tool to improve chickpea growth and its adaptation to salt-degraded soils.

Genomic and transcriptomic analysis of genes involved in exopolysaccharide biosynthesis by Streptococcus thermophilus IMAU20561 grown on different sources of nitrogen.

Exopolysaccharides (EPSs), which are produced by lactic acid bacteria, have been found to improve the texture and functionality of fermented dairy products. In a previous study, four nitrogen sources were identified as affecting the yield, molecular weight and structure of EPSs produced by Streptococcus thermophilus IMAU20561 in M17 medium. In this genomic and transcriptomics study, a novel eps gene cluster responsible for assembly of repeating units of EPS is reported. This eps cluster (22.3 kb), consisting of 24 open reading frames, is located in the chromosomal DNA. To explore the biosynthetic mechanisms in EPS, we completed RNA-seq analysis of S. thermophilus IMAU20561 grown in four different nitrogen sources for 5 h (log phase) or 10 h (stationary phase). GO functional annotation showed that there was a significant enrichment of differentially expressed genes (DEGs) involved in: amino acid biosynthesis and metabolism; ribonucleotide biosynthesis and metabolism; IMP biosynthesis and metabolism; and phosphorus metabolism. KEGG functional annotation also indicated enrichment of DEGs involved in amino acid biosynthesis, glycolysis, phosphotransferase system, fructose, and mannose metabolism. Our findings provide a better understanding the genetic traits of S. thermophilus, the biosynthetic pathways needed for the production of EPS, and a theoretical basis for screening dairy starter cultures.