Exopolysaccharide Research Articles

The synergy of halotolerant PGPB and mauran mitigates salt stress in tomato (Solanum lycopersicum) via osmoprotectants accumulation.

Salinity stress is one of the major abiotic factors limiting sustainable agriculture. Halotolerant plant growth-promoting bacteria (PGPB) increased salt stress tolerance in plants, but the mechanisms underlying the tolerance are poorly understood. This study investigated the PGP activity of four halotolerant bacteria under salinity stress and the tomato salt-tolerance mechanisms induced by the synergy of these bacteria with the exopolysaccharide (EPS) mauran. All PGPB tested in this study were able to offer a significant improvement of tomato plant biomass under salinity stress; Peribacillus castrilensis N3 being the most efficient one. Tomato plants treated with N3 and the EPS mauran showed greater tolerance to NaCl than the treatment in the absence of EPS and PGPB. The synergy of N3 with mauran confers salt stress tolerance in tomato plants by increasing sodium transporter genes' expression and osmoprotectant content, including soluble sugars, polyols, proline, GABA, phenols and the polyamine putrescine. These osmolytes together with the induction of sodium transporter genes increase the osmotic adjustment capacity to resist water loss and maintain ionic homeostasis. These findings suggest that the synergy of the halotolerant bacterium N3 and the EPS mauran could enhance tomato plant growth by mitigating salt stress and could have great potential as an inductor of salinity tolerance in the agriculture sector.

Antimicrobial peptides in combination with citronellal efficiently kills multidrug resistance bacteria.

Antimicrobial peptides (AMPs) are considered as the most potential alternatives to antibiotics, but they have several drawbacks, including high cost, medium antimicrobial efficacy, poor cell selectivity, which limit clinical application. To overcome the above problems, combination therapy of AMPs with adjuvants might maximize the effectiveness of AMPs. We found that citronellal can substantially potentiate the ZY4R peptide efficacy against Escherichia coli ATCC25922. However, it is unclear whether ZY4R/citronellal combination poses synergistic antimicrobial effects against most bacteria, and their synergy mechanism has not been elucidated. To investigate synergistic antimicrobial efficacies, biosafety, and synergy mechanism of ZY4R/citronellal combination. Checkerboard, time-kill curves, cytotoxicity assays, and in vivo animal models were conducted to assess synergistic antimicrobial effects and biosafety of the ZY4R/citronellal combination. To evaluate their synergy mechanism, a series of cell-based assays and transcriptome analysis were performed. ZY4R/citronellal combination exhibited synergistic antimicrobial effects against 20 clinically significant pathogens, with the fractional inhibitory concentration index (FICI) ranging from 0.313 to 0.047. Meanwhile, ZY4R/citronellal combination enhanced antimicrobial efficacies without compromising cell selectivity, contributing to decreasing drug dosage and improving biosafety. Compared with ZY4R (4 mg/kg) and citronellal (25 mg/kg) alone, ZY4R (4 mg/kg)/citronellal (25 mg/kg) combination significantly decreased the bacterial load in peritoneal fluid, liver, and kidney (P < 0.05) and alleviated pathological damage of the organs of mice. Mechanistic studies showed that ZY4R allowed citronellal to pass through the outer membrane rapidly and acted on the inner membrane together with citronellal, causing more potent membrane damage. The membrane damage prompted the continuous accumulation of citronellal in cells, and citronellal further induced energy breakdown and inhibited exopolysaccharide (EPS) production, which aggravated ZY4R-induced outer membrane damage, thereby resulting in bacterial death. ZY4R/citronellal combination exhibited broad-spectrum synergy with a low resistance development and high biosafety. Their synergy mechanism acted on two important cellular targets (energy metabolism and membrane integrity). Combination therapy of ZY4R with citronellal may be a promising mixture to combat bacterial infections facing an antibiotic-resistance crisis.

Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1 eps gene knockout mutants reduced exopolysaccharide synthesis and immunomodulatory activities

In this study, eps genes in Cluster I of Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1 were deleted via conjugal transformation to understand the role of key eps genes in exopolysaccharide (EPS) synthesis and their immunomodulatory effects. Eps gene knockout mutants of OLL1073R-1, ΔepsIB, ΔepsIC, ΔepsID, ΔepsIE, and ΔepsIF were isolated by homologous recombination with the conjugative plasmid pGMβ1. Significant reductions in high molecular weight EPS (H-EPS) production were observed in ΔepsIB, ΔepsIC, and ΔepsIE and partly in ΔepsIF, but not in ΔepsID fermented milk. H-EPS purified from OLL1073R-1 fermented milk showed significant induction of IL-6, IL-10 and IL-12, whereas low molecular weight EPS (L-EPS) showed significant induction of IL-10 and IL-12 in THP-1 dendritic cells. ΔepsIE with less production of H-EPS exhibited significant reductions in IL-6, IL-10 and IL-12 productions. The present study firstly reported that H-EPS in OLL1073R-1 contributes to IL-6, IL-10 and IL-12 production in THP-1 dendritic cells.

Cellulose Synthase in Atacama Cyanobacteria and Bioethanol Production from Their Exopolysaccharides.

Cyanobacteria produce exopolysaccharides (EPSs) as an adaptative mechanism against ultraviolet radiation and desiccation. Cellulose is present in the extracellular polymeric substance in some cyanobacteria genera and it has been proposed as a raw material for biofuel production. The goal of this work was to evaluate the cellulose presence in EPS of Atacama cyanobacteria strains and its use as an alternative and innovative biological source to produce bioethanol. The presence of cellulose was evaluated using techniques of molecular biology, bioinformatics, and electronic microscopy. The conserved motif D,D,D,35QXXRW, characteristic of processive β-glycosyltransferase in all cellulose-producing organisms, was identified in the genome of the LLA-10 strain. This is evidence that cellulose synthase in the LLA-10 strain is a functional enzyme. EPS from Atacama cyanobacteria was hydrolyzed by β-glucosidases (cellobiase and cellulase) and the released glucose was yeast-fermented to ethanol. Ethanol production reached 172.69 ± 0.02 mg ethanol/g EPS after 48 h of incubation. These results are the first step in the evaluation of EPS produced by native cyanobacteria isolated from northern Chile for future biotechnological applications such as the production of bioethanol.

Cyanobacteria as an eco-friendly bioresource for EPS production with crack healing capacity in concrete

Cyanobacteria, alike microalgae, are photosynthetic microorganisms found in different environmental conditions. These photosynthetic bacteria are also considered promising solutions for cracking problem in concretes. These photosynthetic organisms precipitate CaCO3 crystals in concrete microcracks and pores, thus sealing the cracks and increasing the strength gain of the concrete blocks. Cyanobacteria Lyngbya sp., Phormidium sp., and microalgae Chlorella vulgaris were studied initially. Lyngbya sp., Phormidium sp., and Chlorella vulgaris have the additional property of secreting the exopolysaccharide (EPS). EPSs are an important class of biopolymers with great ecological importance. Because of their high charge density and capacity to act as mineralization nucleation sites, cyanobacterial exopolysaccharides are currently receiving more attention. Using the three strains stated above, this study investigates the impact of physiochemical factors on biomass concentration and exopolysaccharide synthesis and creates mathematical models. Using Central Composite Design (CCD), it is possible to study the interactions between the many parameters, including nitrate concentration, starting media pH, light intensity, and temperature. The biochemical characteristics of the leftover biomass are assessed after exopolysaccharide extraction, and because of its high carbohydrate and fat content, it can be used to produce biofuel. The study of checking the microbial crack healing capacity and strength gain of the concrete using these three cyanobacterial strains is being studied. Based on the outcome, the best species was chosen. Further microstructural analyses, such as X-ray diffraction (XRD) and scanning electron microscopy (SEM) were used to support the calcium carbonate precipitation.

The effect of NaCl, pH, and phosphate on biofilm formation and exopolysaccharide production by high biofilm producers of Bacillus strains.

Biofilm formation is an effective survival strategy of plant-associated microorganisms in hostile environments, so the application of biofilm-forming and exopolysaccharide (EPS)-producing beneficial microbes to plants has received more attention in recent years. This study examined the ability of biofilm and EPS production of Bacillus subtilis and Bacillus thuringiensis strains under different NaCl concentrations (0, 50, 100, 200, and 400mmol/L), pH values (5.5, 6.5, 7.5, and 8.5), and phosphate levels (0, 25, 50, and 100mmol/L at 0 and 400mmol/L NaCl). B. subtilis BS2 and B. thuringiensis BS6/BS7 strains significantly increased biofilm formation in a similar pattern to EPS production under salt stress. B. subtilis BS2/BS3 enhanced biofilm production at slightly acidic pH with a lower EPS production but the other strains formed considerably more amount of biofilm and EPS at alkaline pH. Interestingly, higher levels of phosphate substantially decreased biofilm and EPS production at 0mmol/L NaCl but increased biofilm formation at 400mmol/L salt concentration. Overall, contrary to phosphate, salt and pH differently influenced biofilm and EPS production by Bacillus strains. EPS production contributed to biofilm formation to some extent under all the conditions tested. Some Bacillus strains produced more abundant biofilm under salt and pH stress, indicating their potential to form in vivo biofilms in rhizosphere and on plants, particularly under unfavorable conditions.

Effects of fungal carbon dots application on growth characteristics and cadmium uptake in maize

The advancement of nanotechnology has led to the increased use of nanomaterials for the purpose of restoring contaminated soils. However, so far no research has been reported on the interactions of carbon dots with heavy metals (loid)s in phytoremediation. The purpose of this study was to investigate the effect of a new carbon dots derived from fungal exopolysaccharide (EPSs) on the growth and cadmium uptake in maize plants. This research was carried out using a completely randomized design with three replications in a greenhouse condition. Treatments included control, carbon dots (150 mg kg−1), cadmium (50 mg kg−1) and cadmium + carbon dots (50 mg kg−1+150 mg kg−1). The carbon dots synthesized by hydrothermal method from EPSs. The results showed that shoot dry weight and chlorophyll content of maize increased 9.7% and 23.2% in the presence of carbon dots, respectively. Carbon dots improved the chlorophyll content of maize by 24.3% in the cadmium treatment. Cadmium concentration increased (106%) in maize shoot but it decreased in root maize (68%). Carbon dots caused an increase of 5.7 and 6.7 times in the transfer factor and phytoremediation rate of cadmium, respectively. The presence of carbon dots triggered an increase of 77.9% and 39.9% of dissolved organic carbon in non-contaminated and cadmium-contaminated soils, respectively. Soil microbial biomass carbon increased 54.9% and 24.1% carbon dots and cadmium + carbon dots treatments, respectively. The study demonstrates the potential of fungal carbon dots for phytoremediation of heavy metal (loid)s contaminated soils. It also highlights the potential of nanotechnology in environmental remediation efforts.

Comparative study of virulence factors, pathogenicity and genetic diversity of Acidovorax avenae subsp. avenae, the causal agent of red stripe disease in sugarcane

AbstractAcidovorax avenae subsp. avenae (Aaa) is the causal agent of red stripe in sugarcane, a disease characterized by two forms: leaf stripe and top rot. Despite the importance of this disease, little is known about Aaa infection mechanisms, and studies of virulence factors (VFs) have not been reported. Previous analysis of Aaa isolates obtained from sugarcane fields in Tucumán, Argentina, suggested that pathogenicity was not related to genetic groups determined by rep‐PCR analysis. Therefore, the aims of this work were to analyse VFs produced by Aaa strains from different genetic groups and to evaluate the association among VFs, between VFs and Aaa pathogenicity and between VFs and Aaa genetic diversity. Bacterial motility (sliding, swimming and twitching), extracellular enzyme activities (proteases, amylases and endoglucanases), H2O2‐mediated oxidative stress, cell adhesion and exopolysaccharide (EPS) production were assayed in 14 Aaa isolates from different genetic groups. Pearson's correlation analysis and principal component analysis (PCA) were performed. For each VF analysed, values were variable presenting quantitative traits among the strains. Significant correlations were observed among the VFs, though no clear trend was identified between them and the pathogenicity or genetic group. Results reinforce the idea that a single VF does not account for the infection outcome, but a more complex process occurs where the interplay among VFs must be considered. The results of this study are crucial because they provide insight into the infection mechanism of this bacterium.

Genetic and Phenotypic Analysis of Phage-Resistant Mutant Fitness Triggered by Phage-Host Interactions.

The emergence of phage-resistant bacterial strains is one of the biggest challenges for phage therapy. However, the emerging phage-resistant bacteria are often accompanied by adaptive trade-offs, which supports a therapeutic strategy called "phage steering". The key to phage steering is to guide the bacterial population toward an evolutionary direction that is favorable for treatment. Thus, it is important to systematically investigate the impacts of phages targeting different bacterial receptors on the fitness of the bacterial population. Herein, we employed 20 different phages to impose strong evolutionary pressure on the host Pseudomonas aeruginosa PAO1 and examined the genetic and phenotypic responses of their phage-resistant mutants. Among these strains with impaired adsorptions, four types of mutations associated with bacterial receptors were identified, namely, lipopolysaccharides (LPSs), type IV pili (T4Ps), outer membrane proteins (OMPs), and exopolysaccharides (EPSs). PAO1, responding to LPS- and EPS-dependent phage infections, mostly showed significant growth impairment and virulence attenuation. Most mutants with T4P-related mutations exhibited a significant decrease in motility and biofilm formation ability, while the mutants with OMP-related mutations required the lowest fitness cost out of the bacterial populations. Apart from fitness costs, PAO1 strains might lose their resistance to antibiotics when counteracting with phages, such as the presence of large-fragment mutants in this study, which may inspire the usage of phage-antibiotic combination strategies. This work provides methods that leverage the merits of phage resistance relative to obtaining therapeutically beneficial outcomes with respect to phage-steering strategies.

Positive regulation of the PhcB neighbouring regulator PrhX on expression of the type III secretion system and pathogenesis in Ralstonia solanacearum.

Ralstonia solanacearum PhcB and PhcA control a quorum-sensing (QS) system that globally regulates expression of about one third of all genes, including pathogenesis genes. The PhcB-PhcA QS system positively regulates the production of exopolysaccharide (EPS) and negatively regulates hrp gene expression, which is crucial for the type III secretion system (T3SS). Both EPS and the T3SS are essential for pathogenicity. The gene rsc2734 is located upstream of a phcBSR operon and annotated as a response regulator of a two-component system. Here, we demonstrated that RSc2734, hereafter named PrhX, positively regulated hrp gene expression via a PrhA-PrhIR-PrhJ-HrpG signalling cascade. Moreover, PrhX was crucial for R. solanacearum to invade host roots and grow in planta naturally. prhX expression was independent of the PhcB-PhcA QS system. PrhX did not affect the expression of phcB and phcA and the QS-dependent phenotypes, such as EPS production and biofilm formation. Our results provide novel insights into the complex regulatory network of the T3SS and pathogenesis in R. solanacearum.

Exopolysaccharide from Weissella confusa J4-1 inhibits colorectal cancer via induction of cell cycle arrest

Exopolysaccharide (EPS), a bioproduct of lactic acid bacteria (LAB), has various health-promoting biological activities that may be beneficial for cancer therapy. This in vivo and in vitro study aimed to elucidate the anti-colorectal cancer (CRC) capacity of a homopolysaccharide EPS obtained from Weissella confusa J4-1 (EPSJ4-1) isolated from the faeces of healthy infants. We confirmed that EPSJ4-1 contained glucose and effectively suppressed the proliferation, migration, and invasion of CRC cells. EPSJ4-1 treatment significantly retarded the growth of HT-29 tumour xenografts without causing cytotoxicity to normal organs. EPSJ4-1 exerts an inhibitory effect on cell proliferation by inducing G0/G1 phase cell cycle arrest in CRC cells. Furthermore, EPSJ4-1 upregulated p21 levels and downregulated mutant p53 and cyclin kinase 2 levels. This is the first study to demonstrate the antitumour effects of EPS from W. confusa on CRC via cell cycle arrest and inhibition of cell migration and invasion, suggesting that EPSJ4-1 has the potential to be developed as a nutraceutical or pharmaceutical drug to prevent and treat CRC.

Unraveling the molecular mechanisms of selenite reduction: transcriptomic analysis of Bacillus reveals the key role of sulfur assimilation.

Selenite biotransformation by microorganisms is an effective detoxification and assimilation process. However, current knowledge of the molecular mechanisms of selenite reduction remains circumscribed. Here, the reduction of Se(IV) by a highly selenite-resistant Bacillus sp. SL (up to 50mM) was systematically analyzed, and the molecular mechanisms of selenite reduction were investigated. Remarkably, 10mM selenite was entirely transformed by the strain SL within 20h, demonstrating a faster conversion rate compared to other microorganisms. Furthermore, glutathione (GSH) and exopolysaccharides (EPS) changes were also monitored during the process. Transcriptomic analysis revealed that the genes of ferredoxin-sulfite oxidoreductase (6.82) and sulfate adenylyltransferase (6.32) were significantly upregulated, indicating that the sulfur assimilation pathway is the primary reducing pathway involved in selenite reduction by strain SL. Moreover, key genes associated with NAD(P)/FAD-dependent oxidoreductases and thioredoxin were significantly upregulated. The reduction of Se(IV) was mediated by multiple pathways in strain SL. To our knowledge, this is the initial report to identify the involvement of sulfur assimilation pathway in selenite reduction for bacillus, which is rare in aerobic bacteria.

Dietary Administration Effects of Exopolysaccharide Produced by Bacillus tequilensis PS21 Using Riceberry Broken Rice, and Soybean Meal on Growth Performance, Immunity, and Resistance to Streptococcus agalactiae of Nile tilapia (Oreochromis niloticus).

Overuse of antibiotics in aquaculture has generated bacterial resistance and altered the ecology. Aquacultural disease control requires an environmentally sustainable approach. Bacterial exopolysaccharides (EPSs) as bioimmunostimulants have not been extensively explored in aquaculture. This study investigated EPS produced from 5% w/v riceberry broken rice as a carbon source and 1% w/v soybean meal as a nitrogen source by Bacillus tequilensis PS21 from milk kefir grain for its immunomodulatory, antioxidant activities and resistance to pathogenic Streptococcus agalactiae in Nile tilapia (Oreochromis niloticus). The FTIR spectrum of EPS confirmed the characteristic bonds of polysaccharides, while the HPLC chromatogram of EPS displayed only the glucose monomer subunit, indicating its homopolysaccharide feature. This EPS (20 mg/mL) exhibited DPPH scavenging activity of 65.50 ± 0.31%, an FRAP value of 2.07 ± 0.04 mg FeSO4/g DW, and antimicrobial activity (14.17 ± 0.76 mm inhibition zone diameter) against S. agalactiae EW1 using the agar disc diffusion method. Five groups of Nile tilapia were fed diets (T1 (Control) = 0.0, T2 = 0.1, T3 = 0.2, T4 = 1.0, and T5 = 2.0 g EPS/kg diet) for 90 days. Results showed that EPS did not affect growth performances or body composition, but EPS (T4 + T5) significantly stimulated neutrophil levels and serum lysozyme activity. EPS (T5) significantly induced myeloperoxidase activity, catalase activity, and liver superoxide dismutase activity. EPS (T5) also significantly increased the survival of fish at 80.00 ± 5.77% at 14 days post-challenge with S. agalactiae EW1 compared to the control (T1) at 53.33 ± 10.00%. This study presents an efficient method for utilizing agro-industrial biowaste as a prospective source of value-added EPS via a microbial factory to produce a bio-circular green economy model that preserves a healthy environment while also promoting sustainable aquaculture.

Lignan-containing maple products inhibit Listeria monocytogenes biofilms on fresh produce.

Major listeriosis outbreaks have been associated with fresh produce contaminated with Listeria monocytogenes. Strains that synthesize the Pss exopolysaccharide (EPS) have an estimated 102 to 104-fold advantage over nonsynthesizing strains in causing listeriosis. They more readily attach to the surfaces of fruit and vegetables forming EPS-biofilms that better withstand stresses associated with produce storage and consumption. Here, we show that the threat to fresh produce safety posed by the listerial EPS-biofilms may be countered by broadly available maple products. We serendipitously discovered that aqueous extracts of wood from several Acer (maple) and Carya (pecan, hickory) species inhibit the formation of listerial EPS-biofilms without affecting bacterial viability. One active ingredient in maple wood was identified as nortrachelogenin-8'-O-β-D-glucopyranoside (NTG). At 120 μM, this lignan decreased colonization of the EPS-synthesizing L. monocytogenes on cantaloupe pieces by approximately 150-fold, and on cut celery and lettuce by 10 to 11-fold. Another lignan, lariciresinol, which is abundant in a common food sweetener, maple syrup, had antibiofilm activity comparable to that of NTG. Diluted in the range of 1:200 to 1:800 maple syrup from two random manufacturers prevented formation of listeiral EPS-biofilms. Importantly, not only did maple products drastically decrease colonization of fresh produce by the EPS-synthesizing strains, they also decreased, by 6 to 30-fold, colonization by the L. monocytogenes strains that do not synthesize measurable EPS, including strains from the infamous 2011 cantaloupe listeriosis outbreak. Inhibition of surface colonization by various listerial strains, broad availability of maple sap and syrup as well as maple lumber processing waste position maple products as potential antibiofilm agents for protecting fresh produce from L. monocytogenes.

Structural characterization, biofunctionality, and environmental factors impacting rheological properties of exopolysaccharide produced by probiotic Lactococcus lactis C15

Exopolysaccharides (EPSs) possess distinctive rheological and physicochemical properties and innovative functionality. This study aimed to investigate the physicochemical, bioactive, and rheological properties of an EPS secreted by Lactococcus lactis subsp. lactis C15. EPS-C15 was found to have an average molecular weight of 8.8 × 105 Da and was identified as a hetero-EPS composed of arabinose, xylose, mannose, and glucose with a molar ratio of 2.0:2.7:1.0:21.3, respectively. The particle size and zeta potential represented 311.2 nm and − 12.44 mV, respectively. FITR exhibited that EPS-C15 possessed a typical polysaccharide structure. NMR displayed that EPS-C15 structure is → 3)α-d-Glcvi (1 → 3)α-d-Xylv (1 → 6)α-d-Glciv(1 → 4)α-d-Glc(1 → 3)β-d-Man(1 → 2)α-d-Glci(1 → . EPS-C15 scavenged DPPH and ABTS free radicals with 50.3% and 46.4% capacities, respectively. Results show that the antiproliferative activities of EPS-C15 revealed inhibitions of 49.7% and 88.1% against MCF-7 and Caco-2 cells, respectively. EPS-C15 has antibacterial properties that inhibited Staphylococcus aureus (29.45%), Salmonella typhimurium (29.83%), Listeria monocytogenes (30.33%), and E. coli O157:H7 (33.57%). The viscosity of EPS-C15 decreased as the shear rate increased. The rheological properties of the EPS-C15 were affected by changes in pH levels and the addition of salts. EPS-C15 is a promising biomaterial that has potential applications in various industries, such as food, pharmaceuticals, and healthcare.

Physicochemical Characterization of Dextran HE29 Produced by the Leuconostoc citreum HE29 Isolated from Traditional Fermented Pickle.

In this study, lactic acid bacteria (LAB) strains were isolated from traditional fermented pickles, and among the identified strains, Leuconostoc citreum HE29 with a strong slimy colony profile was further selected to determine the physicochemical and techno-functional properties of its exopolysaccharide (EPS). Glucose was the only sugar monomer in the core unit of EPS HE29 detected by HPLC analysis, and glucan HE29 revealed 7.3 kDa of molecular weight. Structural characterization of glucan HE29 by 1H and 13C NMR spectroscopy analysis demonstrated that EPS HE29 was a dextran-type EPS containing 5.3% levels of (1 → 3)-linked α-D-glucose units. This structural configuration was also supported by FT-IR analysis, which also demonstrated the functional groups within the dextran HE29 structure. In terms of thermal properties detected by TGA and DSC analysis, dextran HE29 demonstrated a degradation temperature of around 280 °C, showing its strong thermal features. A semi-crystalline nature was observed for dextran HE29 detected by XRD analysis. Finally, AFM and SEM analysis revealed tangled network-like properties and web-like branched structures for dextran HE29, respectively. These findings suggest the importance of plant-based fermented products as LAB sources in obtaining novel EPS structures with potential techno-functional roles.

Biosynthesis and characterization of antibacterial bacterial cellulose composite membrane composed of montmorillonite and exopolysaccharides

Bacterial cellulose (BC), as a natural renewable polymer material, has the advantages of porous nanonetwork structure, high degree of polymerization, high purity, high crystallinity, excellent mechanical properties and biocompatibility. However, BC lacks antibacterial properties, which leads to the limitation of BC material in food packaging and medical materials. In this study, a new antibacterial material using the combination of montmorillonite (MMT), BC and exopolysaccharides (EPS) produced by Weissella confusa H2 was synthesized. Fourier infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) analysis showed that BC-EPS, BC-MMT and BC-EPS-MMT composite membranes conformed to the typical type I cellulose structure. Compared to BC membrane, scanning electron microscopy (SEM) showed that the porosity of BC-EPS, BC-MMT and BC-EPS-MMT composite membranes was low and compact. The physical properties of BC-EPS, BC-MTT and BC-EPS-MTT composite membranes showed lower water vapor transmittance. The BC-MTT and BC-EPS-MTT composite membranes exhibit a lower swelling ratio in 120 min. The thermal properties show that BC-EPS, BC-MTT and BC-EPS-MTT composite membranes have higher thermal stability (352 °C, 310 °C, 314 °C). Additionally, both BC-MMT and BC-EPS-MMT demonstrated strong inhibitory effects against various bacterial strains, including Staphylococcus aureus, Escherichia coli, Salmonella paratyphi A, and Bacillus subtilis. The exceptional properties exhibited by composite membranes establishes them as a highly promising option in the field of food packaging and medical material applications.

New strategy for bioplastic and exopolysaccharides production: Enrichment of field microbiomes with cyanobacteria

Seven photosynthethic microbiomes were collected from field environmental samples to test their potential in polyhydroxybutyrate (PHB) and exopolysaccharides (EPS) production, both alternatives to chemical-based polymers. Microscope observations together with microbial sequence analysis revealed the microbiome enrichment in cyanobacteria after culture growth under phosphorus limitation. PHB and EPS production were studied under three culture factors (phototrophy, mixotrophy and heterotrophy) by evaluating and optimizing the effect of three parameters (organic and inorganic carbon and days under light:dark cycles) by Box-Behnken design. Results showed that optimal conditions for both biopolymers synthesis were microbiome-dependent; however, the addition of organic carbon boosted PHB production in all the tested microbiomes, producing up to 14 %dcw PHB with the addition of 1.2 g acetate·L−1 and seven days under light:dark photoperiods. The highest EPS production was 59 mg·L−1 with the addition of 1.2 g acetate·L−1 and four days under light:dark photoperiods. The methodology used is suitable for enriching microbiomes in cyanobacteria, and for testing the best conditions for bioproduct synthesis for further scale up.

Synthesis in situ of heteropolysaccharide by Levilactobacillus brevis AM7 during fermentation of oat and hemp and its effect on the techno-functional properties of oat yogurt type model

Plant raw materials, such as oat bran concentrate (OBC), oat flour (OF), and hemp protein concentrate (HP), are significant sources of dietary fibers and proteins with high nutritional quality. Exopolysaccharides (EPS) synthesized by lactic acid bacteria (LAB) during fermentation are natural texturing agents that can replace hydrocolloid additives (e.g., carrageenan, guar gum, and pectin). This study aimed to evaluate EPS synthesis in OBC, OF, and HP during fermentation with Levilactobacillus brevis AM7 supplemented with glucose. Monosaccharide analysis identified that EPS from L. brevis AM7 was a heteropolysaccharide (HePS) consisting of rhamnose, mannose, glucose, and glucose-amine. Rheological analysis revealed that HePS production led to increased viscosity and viscoelastic moduli (G′ and G″) of the fermented materials compared to control. Fermented OBC exhibited the most stable structure among the three materials. Fermented OBC also showed the lowest acidity level, whereas fermented HP exhibited intensive acidification. This was likely due to the more abundant flour endogenous sugars (sucrose, raffinose, and stachyose) in HP and their fast utilization during fermentation. A yogurt alternative product was prepared from fermented OBC milk, showing a high LAB cell density, a pH of 4.2, and a significantly higher viscosity than the chemically acidified control. The in situ synthesized HePS counteracted the viscosity loss and inhibited syneresis of the yogurt alternative product during four days of storage at 4 °C. Overall, the use of L. brevis AM7 producing HePS enabled improved techno-functional properties of plant raw materials and showed potential for application in yogurt alternatives.

Xylella fastidiosa modulates exopolysaccharide polymer length and the dynamics of biofilm development with a β-1,4-endoglucanase.

It is well established that exopolysaccharide (EPS) is an integral structural component of bacterial biofilms necessary for assembly and maintenance of the three-dimensional architecture of the biofilm. However, the process and role of EPS turnover within a developing biofilm is not fully understood. Here, we demonstrated that Xylella fastidiosa uses a self-produced endoglucanase to enzymatically process its own EPS to modulate EPS polymer length. This enzymatic processing of EPS dictates the early stages of X. fastidiosa's biofilm development, which, in turn, affects its behavior in planta. A deletion mutant that cannot produce the endoglucanase was hypervirulent, thereby linking enzymatic processing of EPS to attenuation of virulence in symptomatic hosts, which may be a vestige of X. fastidiosa's commensal behavior in many of its other non-symptomatic hosts.