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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 (Kin D) and response regulatory protein Spo0 A in the phosphoric acid relay system. Key transcriptional regulatory factors, including Omp R 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. [Display omitted] • Salt stress prompts early EPS secretion and enhances Bacillus survival. • Salt stress alters the composition of polysaccharide and protein content in EPS. • Kin D and Spo0 A pathways regulate EPS synthesis under salt stress. • Higher salt boost EPS secretion, elevating viscosity and forming larger aggregates. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Native lactic acid bacteria produced exopolysaccharides from glucose and sucrose. • Exopolysaccharides were produced by lactic acid bacteria using agave by-products. • EPS from agave by-products showed high degradation temperatures. [ABSTRACT FROM AUTHOR]

In the study, in order to develop novel starter cultures, three types of yoghurt were produced with different combinations of autochthonous yoghurt bacteria and the samples were investigated in terms of fermentation, microbiological quality, physicochemical properties (pH, acidity, water-holding capacity, apparent viscosities, and rheological properties), organic and benzoic acid contents, and sensory properties during 28 days. Lactobacillus delbrueckii subsp. bulgaricus content of all samples was between 7.29 and 8.58, and Streptococcus thermophilus content was between 8.03 and 9.02 log CFU/g during storage. Yoghurt type and storage time statistically affected pH, acidity, water-holding capacity, apparent viscosity, rheological properties, organic and benzoic acid content, and some sensory parameters. The general acceptability scores of the probiotic, traditional, exopolysaccharide, and control samples during storage were 8.27, 7.82, 7.53, and 7.47 respectively. Moreover, the results revealed that the strain combinations for probiotic, traditional, and exopolysaccharide yoghurt samples can be used as novel starter cultures for yoghurt production. [ABSTRACT FROM AUTHOR]

Interactions between tumoral cells and tumor‐associated bacteria within the tumor microenvironment play a significant role in tumor survival and progression, potentially impacting cancer treatment outcomes. In lung cancer patients, the Gram‐negative pathogen Pseudomonas aeruginosa raises questions about its role in tumor survival. Here, a microfluidic‐based 3D‐human lung tumor spheroid‐P. aeruginosa model is developed to study the bacteria's impact on tumor survival. P. aeruginosa forms a tumor‐associated biofilm by producing Psl exopolysaccharide and secreting iron‐scavenging pyoverdine, which is critical for establishing a bacterial community in tumors. Consequently, pyoverdine promotes cancer progression by reducing susceptibility to iron‐induced death (ferroptosis), enhancing cell viability, and facilitating several cancer hallmarks, including epithelial–mesenchymal transition and metastasis. A promising combinatorial therapy approach using antimicrobial tobramycin, ferroptosis‐inducing thiostrepton, and anti‐cancer doxorubicin could eradicate biofilms and tumors. This work unveils a novel phenomenon of cross‐kingdom cooperation, where bacteria protect tumors from death, and it paves the way for future research in developing antibiofilm cancer therapies. Understanding these interactions offers potential new strategies for combatting cancer and enhancing treatment efficacy. [ABSTRACT FROM AUTHOR]

Background: Cooling towers (CTs) are crucial to myriad industrial processes, supporting thermal exchange between fluids in heat exchangers using water from lakes and rivers as coolant. However, CT water can sometimes introduce microbial contaminants that adhere to and colonize various surfaces within the CT system. These microorganisms can form biofilms, significantly hindering the system's thermal exchange efficiency. Current treatment strategies employ oxidizing biocides to prevent microbial growth. However, despite their affordability, they do not eliminate biofilms effectively and can lead to corrosive damage within the system. Herein, we aim to devise an anti-biofilm strategy utilizing hydrolytic enzymes (such as α-amylase, glucoamylase, pectin-lyase, cellulase, protease, and DNase) alongside antibiotics (including meropenem, ciprofloxacin, gentamicin, erythromycin, chloramphenicol, and ceftriaxone) to combat microbial growth and biofilm formation in cooling systems. Results: All enzymes reduced the development of the biofilms significantly compared to controls (p < 0.05). The polysaccharidases exhibited biomass reduction of 90%, except for pectin-lyase (80%), followed by DNAse and protease at 43% and 49%, respectively. The antibiotics reduced the biofilms of 70% of isolates in concentration of > 2 mg/mL. The minimal biofilm eradication concentration (MBEC) lower than 1 mg/mL was detected for some 7-day-old sessile isolates. The enzymes and antibiotics were also used in combination against biofilms using the modified Chequerboard method. We found six synergistic combinations, with Fractional inhibitory concentrations (FIC) < 0.5, out of the ten tested. In the presence of the enzymatic mixture, MBECs presented a significant decrease (p < 0.05), at least 4-fold for antibiotics and 32-fold for enzymes. Moreover, we characterized high molecular weight (> 12 kDa) exopolysaccharides (EPS) from biofilms of ten isolates, and glycosyl composition analysis indicated a high frequency of glucose, mannose, erythrose, arabinose, and idose across isolates EPS contrasting with rhamnose, allose, and those carbohydrates, which were detected in only one isolate. Conclusion: The synergistic approach of combining enzymes with antibiotics emerges as a highly effective and innovative strategy for anti-biofilm intervention, highlighting its potential to enhance biofilm management practices. [ABSTRACT FROM AUTHOR]

Autophagy, a lysosome-dependent protein degradation mechanism, is a highly conserved catabolic process seen in all eukaryotes. This cell protection system, which is present in all tissues and functions at a basic level, can be up- or downregulated in response to various stresses. A disruption in the natural route of the autophagy process is frequently followed by an interruption in the inherent operation of the body’s cells and organs. Probiotics are live bacteria that protect the host through various mechanisms. One of the processes through which probiotics exert their beneficial effects on various cells and tissues is autophagy. Autophagy can assist in maintaining host homeostasis by stimulating the immune system and affecting numerous physiological and pathological responses. In this review, we particularly focus on autophagy impairments occurring in several human illnesses and investigate how probiotics affect the autophagy process under various circumstances.Abbreviation: AD: Alzheimer disease; AKT: AKT serine/threonine kinase; AMPK: 5′AMP-activated protein kinase; ATG: autophagy related; CCl4: carbon tetrachloride; CFS: cell-free supernatant; CMA: chaperone-mediated autophagy; CRC: colorectal cancer; EPS: L. plantarum H31 exopolysaccharide; HD: Huntington disease; HFD: high-fat diet; HPV: human papillomavirus; IFNG/IFN-γ: interferon gamma; IL6: interleukin 6; LGG: L. rhamnosus GG; LPS: lipopolysaccharide; MTOR: mechanistic target of rapamycin kinase; MTORC1: MTOR complex 1; NAFLD: non-alcoholic fatty liver disease; NASH: non-alcoholic steatohepatitis; PD: Parkinson disease; Pg3G: pelargonidin-3-O-glucoside; PI3K: phosphoinositide 3-kinase; PolyQ: polyglutamine; ROS: reactive oxygen species; SCFAs: short-chain fatty acids; SLAB51: a novel formulation of lactic acid bacteria and bifidobacteria; Slp: surface layer protein (of acidophilus NCFM); SNCA: synuclein alpha; ULK1: unc-51 like autophagy-activating kinase 1; YB: B. longum subsp. infantis YB0411; YFP: yeast fermentate prebiotic. [ABSTRACT FROM AUTHOR]

Sanghuang as a medicinal fungus in China has a history of more than 2000 years, and is known as the "forest gold". Most notably, the polysaccharides of Sanghuangporus sp. have attracted widespread attention due to their significant bioactivity in recent years. At present, extensive studies are being carried out on the extraction methods, structural characterization, and activity evaluation of polysaccharides. Here, we aimed to evaluate the structure and bioactivity of LEPS-1, an exopolysaccharide derived from the S. sanghuang JM-1 strain. The structure was elucidated by chromatography/spectral methods and hydrolyzation, and the solubility, the antioxidant activity, hypoglycemic activity and immunomodulatory activity were investigated. Results showed that LEPS-1 contained a →2)-α-Manp(1→6)-α-Galp(1→[2)-α-Manp(1→]n→2,6)-α-Manp(1→6,2)-α-Manp(1→3)-α-Manp(1→ backbone substituted at the O-6 and O-2 positions with side chains. These two branching fragments were β-Manp(1→. The molecular weight of LEPS-1 is 36.131 kDa. The results of biological activity analysis suggested that LEPS-1 was easily soluble in water, with reducing capability and DPPH radical scavenging capability. Furthermore, the IC50 values of LEPS-1 against α-amylase and α-glucosidase were 0.96 mg/mL and 1.92 mg/mL. LEPS-1 stimulated RAW264.7 cells to release NO, TNF-α and IL-6 with no cytotoxicity, showing potent potential for immunomodulatory activity. These findings describe a potential natural exopolysaccharide with medicinal value and a basis for the development of S. sanghuang exopolysaccharides. [ABSTRACT FROM AUTHOR]

Bacterial biofilm formation and attachment to hosts are mediated by carbohydrate-binding lectins, exopolysaccharides, and their interactions in the extracellular matrix (ECM). During tomato infection Ralstonia pseudosolanacearum (Rps) GMI1000 highly expresses three lectins: LecM, LecF, and LecX. The latter two are uncharacterized. We evaluated the roles in bacterial wilt disease of LecF, a fucose-binding lectin, LecX, a xylose-binding lectin, and the Rps exopolysaccharide EPS I. Interestingly, single and double lectin mutants attached to tomato roots better and formed more biofilm under static conditions in vitro. Consistent with this finding, static bacterial aggregation was suppressed by heterologous expression of lecFGMI1000 and lecXGMI1000 in other Ralstonia strains that naturally lack these lectins. Crude ECM from a ΔlecF/X double mutant was more adhesive than the wild-type ECM, and LecF and LecX increased Rps attachment to ECM. The enhanced adhesiveness of the ΔlecF/X ECM could explain the double mutant's hyper-attachment in static conditions. Unexpectedly, mutating lectins decreased Rps attachment and biofilm viscosity under shear stress, which this pathogen experiences in plant xylem. LecF, LecX, and EPS I were all essential for biofilm development in xylem fluid flowing through cellulose-coated microfluidic channels. These results suggest that under shear stress, LecF and LecX increase Rps attachment by interacting with the ECM and plant cell wall components like cellulose. In static conditions such as on root surfaces and in clogged xylem vessels, the same lectins suppress attachment to facilitate pathogen dispersal. Thus, Rps lectins have a dual biological function that depends on the physical environment. Author summary: Bacterial wilt diseases caused by Ralstonia species inflict significant losses on diverse, globally important agricultural plants. The pathogen first colonizes roots and ultimately the water-transporting xylem. There it attaches to host cell walls and other bacterial cells to form biofilms that eventually block xylem vessels and disrupt sap flow. It is not well known how Ralstonia spp. modulate attachment, but precise control of both attachment and dispersal is critical for successful host colonization over the disease cycle. Excessive adhesion could trap bacteria in a toxic or nutrient-depleted environment. Conversely, insufficient adhesion in a flowing environment could displace bacteria from an optimal niche. We provide evidence of dual, environment-specific roles of carbohydrate-binding lectins and exopolysaccharide EPS I in Ralstonia pseudosolanacearum (Rps) attachment. In static conditions, which Rps experiences on a host root, two lectins suppress bacterial aggregation and adhesion to roots. However, in flowing conditions, which Rps experiences in healthy xylem vessels, the same two lectins and EPS I are essential for biofilm development. The lectins increase the biofilm viscosity and support colony structural integrity, likely by interacting with polysaccharides in the biofilm matrix. This novel multifunctionality of bacterial lectins reveals how pathogens adapt to a physically dynamic host environment. [ABSTRACT FROM AUTHOR]

Isolation of new plant-derived lactic acid bacteria (LAB) is highly prioritized in developing novel starter cultures for plant-based fermentation. This study explores the diversity of LAB in Danish flowers and their potential use for plant-based food fermentation. A total of 46 flower samples under 34 genera were collected for LAB isolation. By introducing an enrichment step, a total of 61 LAB strains were isolated and identified using MALDI-TOF and 16S rRNA sequencing. These strains represent 24 species across 9 genera, predominantly Leuconostoc mesenteroides, Fructobacillus fructosus, Apilactobacillus ozensis, and Apilactobacillus kunkeei. Phenotypic screening for exo-cellular polysaccharide production revealed that 40 strains exhibited sliminess or ropiness on sucrose-containing agar plates. HPLC analysis confirmed that all isolates produced exo-cellular polysaccharides containing glucose, fructose, or galactose as sugar monomers. Therefore, the strains were glucan, fructan, and galactan producers. The suitability of these strains for plant-based fermentation was characterized by using almond, oat, and soy milk. The results showed successful acidification in all three types of plant-based matrices but only observed texture development in soy by Leuconostoc, Weissella, Lactococcus, Apilactobacillus, and Fructobacillus. The findings highlight the potential of flower-derived LAB strains for texture development in soy-based dairy alternatives. [ABSTRACT FROM AUTHOR]

Summary In this study, a lactic acid bacteria strain isolated from oats was used as a producer to obtain exopolysaccharide (EPS) using MRS agar plates. The findings indicated that the bacterial strain identified as Weissella cibaria (accession number: PP515290) was shown to have good probiotic activity, adapt better to alkaline than acidic conditions, and efficiently produce EPS at concentrations of up to 7.93 g L−1. SEM images showed a smooth surface and compact particle whereas FTIR confirmed the basic functional groups of polysaccharides in the sample. Meanwhile, XRD analysis demonstrated a non‐crystalline amorphous structure and thermal analysis indicated two‐phase thermal decomposition with maximum rate at 210 °C. Subsequently, EPS from W. cibaria was applied as stabiliser in set‐type yogurt (ex situ) in comparison with yogurt using mixed culture with W. cibaria (in situ) and traditional starter culture (control). The results showed that the addition of 2% EPS significantly improved syneresis (14.31%) compared to co‐culture (27.50%) and control (29.80%) as well as increased hardness of yogurt during 2‐week refrigeration. [ABSTRACT FROM AUTHOR]

Bacterial exopolysaccharides (EPS) have emerged as one of the key players in the field of heavy metal-contaminated environmental bioremediation. This study aimed to characterize and evaluate the metal biosorption potential of EPS produced by a novel Psychrobacillus strain, NEAU-3TGS, isolated from an iron ore deposit at Tamra iron mine, northern Tunisia. Genomic and pan-genomic analysis of NEAU-3TGS bacterium with nine validated published Psychrobacillus species was also performed. The results showed that the NEAU-3TGS genome (4.48 Mb) had a mean GC content of 36%, 4,243 coding sequences and 14 RNA genes. Phylogenomic analysis and calculation of nucleotide identity (ANI) values (less than 95% for new species with all strains) confirmed that NEAU-3TGS represents a potential new species. Pangenomic analysis revealed that Psychrobacillus genomic diversity represents an "open" pangenome model with 33,091 homologous genes, including 65 core, 3,738 shell, and 29,288 cloud genes. Structural EPS characterization by attenuated total reflectance-Fourier transform infrared (ATRFTIR) spectroscopy showed uronic acid and α-1,4-glycosidic bonds as dominant components of the EPS. X-ray diffraction (XRD) analysis revealed the presence of chitin, chitosan, and calcite CaCO3 and confirmed the amorphous nature of the EPS. Heavy metal bioabsorption assessment showed that iron and lead were more adsorbed than copper and cadmium. Notably, the optimum activity was observed at 37°C, pH=7 and after 3 h contact of EPS with each metal. Genomic insights on iron acquisition and metabolism in Psychrobacillus sp. NEAU-3TGS suggested that no genes involved in siderophore biosynthesis were found, and only the gene cluster FeuABCD and trilactone hydrolase genes involved in the uptake of siderophores, iron transporter and exporter are present. Molecular modelling and docking of FeuA (protein peptidoglycan siderophore-binding protein) and siderophores ferrienterobactine [Fe+3 (ENT)]-3 and ferribacillibactine [Fe+3 (BB)]-3 ligand revealed that [Fe+3 (ENT)]-3 binds to Phe122, Lys127, Ile100, Gln314, Arg215, Arg217, and Gln252. Almost the same for [Fe+3 (ENT)]-3knowledge, this is the first report on the characterization of EPS and the adsorption of heavy metals by Psychrobacillus species. The heavy metal removal capabilities may be advantageous for using these organisms in metal remediation. [ABSTRACT FROM AUTHOR]

Exopolysaccharides (EPSs) are a diverse class of biopolymers synthesized by microorganisms under environmental stress conditions, such as pH, temperature, light intensity, and salinity. They offer biodegradable and environmentally friendly alternatives to synthetic polymers. Their structural versatility and functional properties make them unique in various industries, including food, pharmaceuticals, biomedicine, cosmetics, textiles, petroleum, and environmental remediation. In this way, among the well-known EPSs, homopolysaccharides like dextran, bacterial cellulose, curdlan, and levan, as well as heteropolysaccharides like xanthan gum, alginate, gellan, and kefiran, have found widespread applications in numerous fields. However, recent attention has focused on the potential role of extremophile bacteria in producing EPSs with novel and unusual protective and biological features under extreme conditions. Therefore, this review provides an overview of the functional properties and applications of the commonly employed EPSs. It emphasizes their importance in various industries and scientific endeavors while highlighting the raised interest in exploring EPSs with novel compositions, structures, and properties, including underexplored protective functionalities. Nevertheless, despite the potential benefits of EPSs, challenges persist. Hence, this review discusses these challenges, explores opportunities, and outlines future directions, focusing on their impact on developing innovative, sustainable, and functional materials. [ABSTRACT FROM AUTHOR]

Microbial exopolysaccharides (EPSs) are receiving growing interest today, owing to their diversity in chemical structure and source, multiple functions, and immense potential applications in many food and non-food industries. Their health-promoting benefits for humans deserve particular attention because of their various biological activities and physiological functions. The aim of this paper is to provide a comprehensive review of microbial EPSs, covering (1) their chemical and biochemical diversity, including composition, biosynthesis, and bacterial sources belonging mainly to lactic acid bacteria (LAB) or probiotics; (2) their technological and analytical aspects, especially their production mode and characterization; (3) their biological and physiological aspects based on their activities and functions; and (4) their current and future uses in medical and pharmaceutical fields, particularly for their prebiotic, anticancer, and immunobiotic properties, as well as their applications in other industrial and agricultural sectors. [ABSTRACT FROM AUTHOR]

Fructilactobacillus sanfranciscensis, the dominant species of lactic acid bacteria in sourdoughs, impacts the microstructure and flavor of steamed bread through exopolysaccharide production, acidification, proteolysis, and volatile compound generation. The aim of this study is to investigate the phenotypic diversity and technological traits of 28 F. sanfranciscensis strains of different genotypes isolated from Chinese traditional sourdoughs. The results showed that F. sanfranciscensis strains exhibited substantial variation in proteinase and peptidase activities and the amount of acidification and volatiles in fermented sourdoughs. However, we observed no significant differences in exopolysaccharide production among the strains. The strains Sx14 and Ts1 were further chosen for transcriptomics to gain a deep insight into their intraspecies diversity in sourdough fermentation. Significant transcriptome differentiations between these two strains after 12 h fermentation in sourdoughs were revealed. According to the results, the strain Sx14 possessed higher dipeptidase and aminopeptidase activities, galactose utilization, and lactic and acetic acid production abilities, whereas Ts1 showed higher transmembrane transport of substrates and fructose utilization. [ABSTRACT FROM AUTHOR]

Salinity was considered to have effects on the characteristics, performance microbial communities of aerobic granular sludge. This study investigated granulation process with gradual increase of salt under different gradients. Two identical sequencing batch reactors were operated, while the influent of Ra and Rb was subjected to stepwise increments of NaCl concentrations (0–4 g/L and 0–10 g/L). The presence of filamentous bacteria may contribute to granules formed under lower salinity conditions, potentially leading to granules fragmentation. Excellent removal efficiency achieved in both reactors although there was a small accumulation of nitrite in Rb at later stages. The removal efficiencies of chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP) in Ra were 95.31%, 93.70% and 88.66%, while the corresponding removal efficiencies in Rb were 94.19%, 89.79% and 80.74%. Salinity stimulated extracellular polymeric substances (EPS) secretion and enriched EPS producing bacteria to help maintain the integrity and stability of the aerobic granules. Heterotrophic nitrifying bacteria were responsible for NH 4 +-N and NO 2 −-N oxidation of salinity systems and large number of denitrifying bacteria were detected, which ensure the high removal efficiency of TN in the systems. [ABSTRACT FROM AUTHOR]

The present study investigates the low temperature tolerance strategies of thermophilic bacterium Anoxybacillus rupiensis TPH1, which grows optimally at 55 °C , by subjecting it to a temperature down-shift of 10 °C (45 °C) for 4 and 6 h followed by studying its growth, morphophysiological, molecular and proteomic responses. Results suggested that although TPH1 experienced increased growth inhibition, ROS production, protein oxidation and membrane disruption after 4 h of incubation at 45 °C yet maintained its DNA integrity and cellular structure through the increased expression of DNA damage repair and cell envelop synthesizing proteins and also progressively alleviated growth inhibition by 20% within two hours i.e., 6 h, by inducing the expression of antioxidative enzymes, production of unsaturated fatty acids, capsular and released exopolysaccharides and forming biofilm along with chemotaxis proteins. Conclusively, the adaptation of Anoxybacillus rupiensis TPH1 to lower temperature is mainly mediated by the synthesis of large numbers of defense proteins and exopolysaccharide rich biofilm formation. [ABSTRACT FROM AUTHOR]

An exopolysaccharide (EPS)-producing strain, identified as Weissella cibaria HDL-4, was isolated from litchi. After separation and purification, the structure and properties of HDL-4 EPS were characterized. The molecular weight of HDL-4 EPS was determined to be 1.9 × 10⁶ Da, with glucose as its monosaccharide component. Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR) analyses indicated that HDL-4 EPS was a D-glucan with α-(1→6) and α-(1→4) glycosidic bonds. X-ray diffraction (XRD) analysis revealed that HDL-4 EPS was amorphous. Scanning electron microscope (SEM) and atomic force microscope (AFM) observations showed that HDL-4 EPS possesses pores, irregular protrusions, and a smooth layered structure. Additionally, HDL-4 EPS demonstrated significant thermal stability, remaining stable below 288 °C. It exhibited a strong metal ion adsorption activity, emulsification activity, antioxidant activity, and water-retaining property. Therefore, HDL-4 EPS can be extensively utilized in the food and pharmaceutical industries as an additive and prebiotic. [ABSTRACT FROM AUTHOR]

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight in rice. Polyhydroxyalkanoates (PHAs) consitute a diverse group of biopolyesters synthesized by bacteria under nutrient-limited conditions. The phaC gene is important for PHA polymerization. We investigated the effects of phaC gene mutagensis in Xoo strain PXO99A. The phaC gene knock-out mutant exhibited reduced swarming ability relative to that of the wild-type. Under conditions where glucose was the sole sugar source, extracellular polysaccharide (EPS) production by ΔphaC declined by 44.8%. ΔphaC showed weak hypersensitive response (HR) induction in the leaves of non-host Nicotiana tabacum, concomitant with downregulation of hpa1 gene expression. When inoculated in rice leaves by the leaf-clipping method, ΔphaC displayed reduced virulence in terms of lesion length compared with the wild-type strain. The complemented strain showed no significant difference from the wild-type strain, suggesting that the deletion of phaC in Xoo induces significant alterations in various physiological and biological processes. These include bacterial swarming ability, EPS production, transcription of hrp genes, and glucose metabolism. These changes are intricately linked to the energy utilization and virulence of Xoo during plant infection. These findings revealed involvement of phaC in Xoo is in the maintaining carbon metabolism by functioning in the PHA metabolic pathway. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the strain‐specific fermentative behaviour and metabolite footprint of four Lactobacillus spp., in 48‐h fermented milks. Technological properties (net cell growth, pH and lactic acid production), exopolysaccharides (EPS), short‐chain fatty acids (SCFAs), α‐amino acids and peptides <3 kDa were determined and evaluated by chemometrics. Inter‐strain differences were attributed to technological properties, SCFAs and D‐mannose/galactose/arabinose production. Three clusters (technological properties, EPS and SCFA production) were confirmed by Pearson's correlation and hierarchical clustering and principal component analysis explained 61% of LAB's differential fermentative behaviour. Post hoc chemometric analysis is useful to select LAB with better post‐ and paraprobiotic potential. [ABSTRACT FROM AUTHOR]

Summary: 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. [ABSTRACT FROM AUTHOR]

The exopolymer (ESPp) was obtained from Bacillus licheniformis IDN-EC, composed of a polyglutamic acid and polyglycerol phosphate chain O-substituted with αGal moieties (αGal/αGlcNH2 3:1 molar ratio) and with a 5000 Da molecular weight. The cytotoxicity activity of EPSp was determined by reducing the MTT (3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyltetrazolium bromide) to formazan on HeLa cells. This EPS did not show cytotoxicity against the tested cell line. The ESPp presented great advantages as an antioxidant with free radical scavenging activities (1,1-diphenyl-2-picryl-hydrazyl radical (DPPH),hydroxyl radical (OH), and superoxide anion (O2−)) (65 ± 1.2%, 98.7 ± 1.9%, and 97 ± 1.7%), respectively. Moreover, EPSp increased the enzyme activity for catalase (CAT) and glutathione peroxidase (GSH-Px) in HeLa cells (CAT, 2.6 ± 0.24 U/mL; and GSH-Px, 0.75 ± 0.3 U/L). The presence of ESPp showed a significant protective effect against H2O2 in the cell line studied, showing great viability (91.8 ± 2.8, 89.9 ± 2.9, and 93.5 ± 3.6%). The EPSp presented good emulsifying activity, only for vegetable oils, olive oil (50 ± 2.1%) and sesame (72 ± 3%). Sesame was effective compared to commercials products, Triton X-100 (52.38 ± 1.6%), Tween 20 (14.29 ± 1.1%), and sodium dodecyl sulphate (SDS) (52.63 ± 1.6%). Furthermore, the EPS produced at 0.6 M has potential for environmental applications, such as the removal of hazardous materials by emulsification whilst resulting in positive health effects such as antioxidant activity and non-toxicity. EPSp is presented as a good exopolysaccharide for various applications. [ABSTRACT FROM AUTHOR]

Mining-related lead (Pb) pollution of the soil poses serious hazards to ecosystems and living organisms, including humans. Improved heavy metal phytoremediation efficacy, achieved by using phytostabilizing plants assisted by plant-growth-promoting (PGP) microorganisms, has been presented as an effective strategy for remediating polluted soils. The objective of this research was to examine the response and potential of the plant-growth-promoting bacterium LMR356, a Rhodococcus qingshengii strain isolated from an abandoned mining soil, under lead stress conditions. Compared to non-contaminated culture media, the presence of lead induced a significant decrease in auxin production (from 21.17 to 2.65 μg mL−1) and phosphate solubilization (from 33.60 to 8.22 mg L−1), whereas other PGP traits increased drastically, such as 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity (from 38.17 to 71.37 nmol mg−1 h−1 α-ketobutyrate), siderophore production (from 69 to 83%), exopolysaccharide production (from 1952.28 to 3637.72 mg mL−1), biofilm formation, and motility. We, therefore, investigated the behavior of Sulla spinosissima L. in the presence or absence of this strain under a variety of experimental conditions. Under hydroponic conditions, Sulla plants showed endurance to varying lead concentrations (500–1000 μM). Inoculation of plants with Rhodococcus qingshengii strain LMR356 enhanced plant tolerance, as demonstrated by the increase in plant biomass (ranging from 14.41 to 79.12%) compared to non-inoculated Pb-stressed and non-stressed control plants. Antioxidant enzyme activities (increasing by −42.71 to 126.8%) and chlorophyll (383.33%) and carotenoid (613.04%) content were also augmented. In addition to its impact on plant lead tolerance, strain LMR356 showed a growth-promoting effect on Sulla plants when cultivated in sterilized non-contaminated sand. Parameters such as plant biomass (16.57%), chlorophyll (24.14%), and carotenoid (30%) contents, as well as ascorbate peroxidase (APX), peroxidase (POD), and catalase (CAT) activities, were all elevated compared to non-inoculated plants. Furthermore, when the same plant species was cultivated in highly polluted soil, inoculation increased plant biomass and improved its physiological properties. These findings demonstrate that LMR356 is a phytobeneficial bacterial strain capable of enhancing Sulla growth under normal conditions and improving its heavy metal tolerance in multi-polluted soils. Thus, it can be considered a promising biofertilizer candidate for growing Sulla spinosissima L. or other selected plants intended for application in restoration and stabilization initiatives aimed at reviving and safeguarding environmentally compromised and polluted soils after mining activities. [ABSTRACT FROM AUTHOR]

AbstractThis study aimed to enhance the extracellular polymeric substances (EPS) production of Virgibacillus dokdonensis VITP14 and explore its antioxidant potential. EPS and biomass production by VITP14 strain were studied under different culture parameters and media compositions using one factor at a time method. Among different nutrient sources, glucose and peptone were identified as suitable carbon and nitrogen sources. Furthermore, the maximum EPS production was observed at 5% of inoculum size, 5 g/L of NaCl, and 96 h of fermentation. Response surface methodology was employed to augment EPS production and investigate the optimal levels of nutrient sources with their interaction. The strain was observed to produce actual maximum EPS of about 26.4 g/L for finalized optimum medium containing glucose 20 g/L, peptone 10 g/L, and NaCl 50 g/L while the predicted maximum EPS was 26.5 g/L. There was a nine fold increase in EPS production after optimization study. Additionally, EPS has exhibited significant scavenging, reducing, and chelating potential (>85%) at their higher concentration. This study imparts valuable insights into optimizing moderately halophilic bacterial EPS production and evaluating its natural antioxidant properties. According to findings, V. dokdonensis VITP14 was a promising isolate that will provide significant benefits to biopolymer producing industries. [ABSTRACT FROM AUTHOR]

Probiotic bacteria are distinguished by their ability to produce various functional postbiotic metabolites. Therefore, this study aimed to explore the in vitro antioxidant, antidiabetic, and antibacterial properties of two postbiotics generated by Lactiplantibacillus plantarum O7S1 ( Lpb. plantarum O7S1) during the fermentation process: cell-free supernatant (CFS) and exopolysaccharides (EPS). The antioxidant potential of these postbiotics was assessed using various radical scavenging assays and ferric-reducing antioxidant potential. The antidiabetic activity was evaluated through α-amylase inhibitory assays, while antibacterial activity was determined using agar well diffusion assays. The results of the present study revealed that CFS exhibited significant antioxidant and antidiabetic efficacy in contrast to EPS ( P < 0.001). Specifically, CFS displayed remarkable scavenging ability against DPPH, hydroxyl, and superoxide radicals, with inhibition rates of 88.78, 78.91, and 34.85%, respectively, while EPS showed comparatively lower inhibition rates. Additionally, CFS demonstrated higher reducing activity (0.30 optical density units at 700 nm) and potent α-amylase inhibitory activity (95.87%) compared to EPS (67.17%) ( P < 0.001). The agar well diffusion assay reported that CFS showed significant antimicrobial activity against both Gram-positive and Gram-negative pathogens, while no activity was observed with EPS. Furthermore, carbohydrate fermentation analysis indicated the strain's ability to metabolize various carbohydrates and their derivatives, potentially enhancing digestive health. These findings suggest that both CFS and EPS exhibit promising hypoglycemic, antioxidant, and antibacterial properties, making them potential candidates for incorporation into functional foods and pharmaceuticals aimed at preventing oxidative damage, diabetes, and pathogenic bacterial infections. [ABSTRACT FROM AUTHOR]

Microbial exopolysaccharides (EPS) are extracellular polymeric substances synthesized by bacteria, fungi or algae which are now one of the important food ingredients. In this study, glucan EPS was isolated from lactic acid bacteria (LAB) of fermented food origin and was incorporated in yoghurt to enhance its techno-functional attributes and rheological properties. Glucan EPS concentration at 1% showed significant improvement in syneresis, wheying off, water-holding capacity, and sensorial properties of yoghurt. The water holding capacity (WHC) increased from 47% to about 63% while syneresis decreased from 52% to nearly 30%. Rheological features such as viscosity, storage modulus, loss modulus, and firmness were significantly higher for 1% EPS containing yoghurt (1% ECY) than that of the control. The average particle size of 1% ECY was increased from 2.35 µm to 3.46 µm. The microstructure of EPS containing yoghurt was studied by scanning electron microscopy and confocal laser scanning microscopy. EPS supports the growth of probiotic microflora of yoghurt, which was revealed by analyzing the microbial count of yoghurt. The antioxidant activity as 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (mg Trolox/100 g sample) and reducing power assay (mg ascorbic acid/100 g sample) for 1% ECY and control yoghurt were found to be 27.9 (mg Trolox/100 g sample), 6.44 (mg Trolox/100 g sample) and 0.25 (mg ascorbic acid/100 g sample), 0.19 (mg ascorbic acid/100 g sample), respectively. Significant enhancement of these bio-functional attributes of EPS containing yoghurt makes the glucan EPS a suitable hydrocolloid to develop various functional foods. [ABSTRACT FROM AUTHOR]

Previously, the exopolysaccharides produced by Lactiplantibacillus plantarum C7 isolated from the broiler intestine have been shown to possess probiotic potential. In this study, we highlighted the capacity of this strain to produce exopolysaccharide (EPS) endowed with several biological activities. The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) free radical scavenging activities of EPS were found to be 30.4% and 68.165%, respectively, at 100 mg/mL. The ferrous reducing potential of EPS was measured to be 25.26%. Furthermore, EPS exhibited antibacterial effects against both Gram-negative and Gram-positive pathogens, including Escherichia coli ATCC 10536 and Bacillus cereus ATCC 11778, with inhibition zones of 22.5 ± 0.70 mm and 20 ± 1.41, respectively. On the other hand, the exopolysaccharide also exhibited excellent inhibitory activity against butyrylcholinesterase (BChE), achieving 70.99% inhibition at a concentration of 10 mg/mL. FTIR spectra were used to characterize functional groups in EPS. Our findings proved that EPS from Lactiplantibacillus plantarum C7 could be explored for various applications, particularly in the health and functional food sectors. [ABSTRACT FROM AUTHOR]

This study aimed to enhance extracellular polysaccharide (EPS) production in Cordyceps militaris by constructing a quorum sensing (QS) system to regulate the expression of biosynthetic enzyme genes, including phosphoglucomutase, hexokinase, phosphomannomutase, polysaccharide synthase, and UDP‐glucose 4‐epimerase genes. The study found higher EPS concentrations in seven recombinant strains compared to the wild‐type C. militaris, indicating that the overexpression of key enzyme genes increased EPS production. Among them, the CM‐pgm‐2 strain exhibited the highest EPS production, reaching a concentration of 3.82 ± 0.26 g/L, which was 1.52 times higher than the amount produced by the wild C. militaris strain. Additionally, the regulatory effects of aromatic amino acids on the QS system of the CM‐pgm‐2 strain were investigated. Under the influence of 45 mg/L tryptophan, the EPS production in CM‐pgm‐2 reached 4.75 ± 0.20 g/L, representing a 1.90‐fold increase compared to wild C. militaris strains. This study provided an effective method for the large‐scale production of EPSs in C. militaris, and opened up new avenues for research into fungal QS mechanisms. [ABSTRACT FROM AUTHOR]

The exopolysaccharides produced by Streptococcus thermophilus during its growth can improve the texture of fermented dairy products and have various physiological and biochemical functions. To explore the regulatory mechanism of histidine, isoleucine, and glutamate on the exopolysaccharide of S. thermophilus 937, this study analyzed the exopolysaccharide synthesis pathway of the strain through whole genome sequencing. It then explored the effects of the concentration of 3 amino acids on the strain growth, the production and properties of exopolysaccharides, and the transcription level of epsABCD of the strain. Results showed that the strain had a specific PTS for glucose and sucrose transport system and lactose osmotic enzyme. It had the potential to synthesize UDP-glucose, UDP-galactose, and dTDPrhamnose. There was the eps gene cluster composed of 18 coding genes on the chromosome. When the concentration of 3 amino acids was increased to 15 mmol/L, the exopolysaccharide production of the strain increased by 1.5 times, the molecular weight increased by 1.2 times, the molar ratio of monosaccharide composition changed, and the epsABCD transcription level was significantly upregulated (P < 0.05). The 3 amino acids enhanced the production of exopolysaccharides and regulated the properties by regulating the expression of epsABCD. The results of this study provided a better understanding of the exopolysaccharide biosynthesis of Streptococcus thermophilus at the gene level. [ABSTRACT FROM AUTHOR]

[Display omitted] • High sugar, protein of sludge shows outstanding biodrying potential. • Low ventilation rate improved bio-drying cycle and water removal effect. • Revealed the contribution of different water types to water removal. • Lipid degradation role: Crucial for releasing and transforming bound water. • Energy-saving ventilation changes the degradation sequence of organic matter. The effectiveness of dehydration and utilization processes for citric acid dewatered sludge is hampered by its high concentrations of polysaccharides, proteins, and water-binding properties of microbial extracellular polymers (EPS). This research explores the efficacy and mechanisms involved in extracting water from this type of sludge using biological drying technology, with varying rates of ventilation. Especially pertinent was the use of low ventilation rates as control variables. Our results suggest that a scheduled intermittent ventilation at lower rates allows for the most efficient removal of water, achieving a rate of 41.71 % within eight days, according to the zero-order kinetic model. Remarkably, the peak temperature registered was 60 °C, reaching this threshold in just 0.1 days and maintaining high temperatures for approximately 5.9 days. Component analysis of organic matter illustrated a preferential degradation process for lipids under these ventilation conditions which is pivotal for releasing and transforming bound water for efficient extraction, as well as facilitating the breakdown of easily hydrolysable materials. Further, polysaccharide/protein (EPS) decomposition contributed to water removal, though less significantly. The periodic ventilation strategy allowed for the maximum cumulative temperature to be sustained, demonstrating superior efficiency in harnessing bio-generated heat (82.77 % for water evaporation), resulting in dry sludge suitable for self-sustained combustion at relatively low cost ($26.61/t). Highlighted by this study is the considerable potential of energy-efficient ventilation methods in the biological drying treatment of citric acid fermented sludge and similar industrial waste materials. [ABSTRACT FROM AUTHOR]

Linear, unbranched (1,3;1,4)‐β‐glucans (mixed‐linkage glucans or MLGs) are commonly found in the cell walls of grasses, but have also been detected in basal land plants, algae, fungi and bacteria. Here we show that two family GT2 glycosyltransferases from the Gram‐positive bacterium Sarcina ventriculi are capable of synthesizing MLGs. Immunotransmission electron microscopy demonstrates that MLG is secreted as an exopolysaccharide, where it may play a role in organizing individual cells into packets that are characteristic of Sarcina species. Heterologous expression of these two genes shows that they are capable of producing MLGs in planta, including an MLG that is chemically identical to the MLG secreted from S. ventriculi cells but which has regularly spaced (1,3)‐β‐linkages in a structure not reported previously for MLGs. The tandemly arranged, paralogous pair of genes are designated SvBmlgs1 and SvBmlgs2. The data indicate that MLG synthases have evolved different enzymic mechanisms for the incorporation of (1,3)‐β‐ and (1,4)‐β‐glucosyl residues into a single polysaccharide chain. Amino acid variants associated with the evolutionary switch from (1,4)‐β‐glucan (cellulose) to MLG synthesis have been identified in the active site regions of the enzymes. The presence of MLG synthesis in bacteria could prove valuable for large‐scale production of MLG for medical, food and beverage applications. [ABSTRACT FROM AUTHOR]

This study demonstrates that Lactobacillus can produce exopolysaccharides (EPSs) using alternative carbon sources, such as sugarcane molasses and glycerol. After screening 22 strains of Lactobacillus to determine which achieved the highest production of EPS based on dry weight at 37 °C, the strain Ke8 (L. casei) was selected for new experiments. The EPS obtained using glycerol and glucose as carbon sources was classified as a heteropolysaccharide composed of glucose and mannose, containing 1730 g.mol−1, consisting of 39.4% carbohydrates and 18% proteins. The EPS obtained using molasses as the carbon source was characterized as a heteropolysaccharide composed of glucose, galactose, and arabinose, containing 1182 g.mol−1, consisting of 52.9% carbohydrates and 11.69% proteins. This molecule was characterized using Size Exclusion Chromatography (HPLC), Gas chromatography–mass spectrometry (GC-MS), Fourier-transform infrared spectroscopy (FTIR), and proton nuclear magnetic resonance spectroscopy (1H-NMR). The existence of polysaccharides was confirmed via FT-IR and NMR analyses. The results obtained suggest that Lacticaseibacillus casei can grow in media that use alternative carbon sources such as glycerol and molasses. These agro-industry residues are inexpensive, and their use contributes to sustainability. The lack of studies regarding the use of Lacticaseibacillus casei for the production of EPS using renewable carbon sources from agroindustry should be noted. [ABSTRACT FROM AUTHOR]

Ulva rigida green macroalgae contain a variety of polysaccharides. A recent study investigated the optimum concentration and yield of polysaccharide extraction from oven-dried U. rigida biomass using a water-soluble polysaccharide extraction method that adhered to safety standards. This study utilised complete factorial experiments to examine the effects of varying factors on polysaccharide extraction. Results showed a positive correlation between increased levels of all factors and higher polysaccharide extraction yield. This study also found that the main factors and their interaction had a significant impact on the extracted polysaccharides from U. rigida. The highest polysaccharide content and yield were 9.5 mg/mL and 189 mg/g, respectively. Water-soluble polysaccharides demonstrated the presence of reducing sugar (8 mg/g), phenolics (0.69 mg/g) and flavonoids (1.42 mg/g) and exhibited antioxidant properties. Results revealed that freeze-dried polysaccharide powders were primarily composed of the monosaccharide rhamnose. Preliminary results on the effect of these powders on probiotics demonstrated that supplementation of polysaccharides from U. rigida promoted viable Lactobacillus rhamnosus ATCC 53103 growth during cultivation. This discovery has the potential to revolutionise the human food industry and promote the development of functional ingredients for novel and future food products, with numerous applications in the nutraceutical and pharmaceutical industries. [ABSTRACT FROM AUTHOR]

Background: This 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. Results: The 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. Conclusions: The 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. [ABSTRACT FROM AUTHOR]

In this study, we characterize the exopolymer produced by Halomonas sp. strain TGOS-10 –one of the organisms found enriched in sea surface oil slicks during the Deepwater Horizon oil spill. The polymer was produced during the early stationary phase of growth in Zobell's 2216 marine medium amended with glucose. Chemical and proton NMR analysis showed it to be a relatively monodisperse, high-molecular-mass (6,440,000 g/mol) glycoprotein composed largely of protein (46.6% of total dry weight of polymer). The monosaccharide composition of the polymer is typical to that of other marine bacterial exopolymers which are generally rich in hexoses, with the notable exception that it contained mannose (commonly found in yeast) as a major monosaccharide. The polymer was found to act as an oil dispersant based on its ability to effectively emulsify pure and complex oils into stable oil emulsions—a function we suspect to be conferred by the high protein content and high ratio of total hydrophobic nonpolar to polar amino acids (52.7:11.2) of the polymer. The polymer's chemical composition, which is akin to that of other marine exopolymers also having a high protein-to-carbohydrate (P/C) content, and which have been shown to effect the rapid and non-ionic aggregation of marine gels, appears indicative of effecting marine oil snow (MOS) formation. We previously reported the strain capable of utilising aromatic hydrocarbons when supplied as single carbon sources. However, here we did not detect biodegradation of these chemicals within a complex (surrogate Macondo) oil, suggesting that the observed enrichment of this organism during the Deepwater Horizon spill may be explained by factors related to substrate availability and competition within the complex and dynamic microbial communities that were continuously evolving during that spill. [ABSTRACT FROM AUTHOR]

Introduction: This study used Bacillus amyloliquefaciens DGL1 isolated from the arid sandy land of the Qinghai-Tibetan Plateau as the research strain and investigated the effects of DGL1 on the biomass, physiology, and metabolites of Medicago sativa under different intensities of drought stress to provide a high- quality bacterial source and a theoretical basis for the research and development of biological fertilizer suitable for arid areas. Methods: The exopolysaccharides (EPS), 1-Aminocyclopropane-1-carboxylate deaminase (ACC), and phosphorus solubilizing capacity of DGL1 were determined. The effects of a DGL1 suspension on alfalfa biomass, physiological indexes, degree of peroxidation of cell membranes, and activity of antioxidant enzymes were determined after irrigating roots under drought stress. The effects on soil physicochemical properties were also evaluated, and metabolomics analysis was performed to explore the effect of DGL1 on the metabolites of alfalfa under drought stress. Results: Strain DGL1 produced extracellular polysaccharide EPS and ACC deaminase and was capable of phosphorus solubilization. Treatment with DGL1 increased the biomass of alfalfa under different degrees of drought stress, significantly increased the activities of alfalfa antioxidant enzymes Super Oxide Dismutase (SOD), Peroxidase (POD), and catalase (CAT), reduced the content of MDA and H2O2, and increased the content of quick-acting phosphorus, quick-acting potassium, ammonium nitrogen, and nitrate nitrogen in the soil, thus improving soil fertility. Through metabolomics analysis, DGL1 was shown to affect amino acid metabolic pathways, such as arginine, leucine, glutamate, and tyrosine, as well as the levels of energy-providing polysaccharides and lipids, in alfalfa under 15% PEG-6000 drought stress, enhancing alfalfa's capacity to resist drought stress. Discussion: Strain DGL1 enhances the drought suitability of alfalfa and has the potential for dryland development as a biological agent. [ABSTRACT FROM AUTHOR]

Spider silk is a promising material with great potential in biomedical applications due to its incredible mechanical properties and resistance to degradation of commercially available bacterial strains. However, little is known about the bacterial communities that may inhabit spider webs and how these microorganisms interact with spider silk. In this study, we exposed two exopolysaccharide-secreting bacteria, isolated from webs of an orb spider, to major ampullate (MA) silk from host spiders. The naturally occurring lipid and glycoprotein surface layers of MA silk were experimentally removed to further probe the interaction between bacteria and silk. Extensibility of major ampullate silk produced by Triconephila clavata that was exposed to either Microbacterium sp. or Novosphigobium sp. was significantly higher than that of silk that was not exposed to bacteria (differed by 58.7%). This strain-enhancing effect was not observed when the lipid and glycoprotein surface layers of MA silks were removed. The presence of exopolysaccharides was detected through NMR from MA silks exposed to these two bacteria but not from those without exposure. Here we report for the first time that exopolysaccharide-secreting bacteria inhabiting spider webs can enhance extensibility of host MA silks and silk surface layers play a vital role in mediating such effects. [ABSTRACT FROM AUTHOR]

This article provides detailed information about the threat of healthcare-acquired infections, specifically those caused by the Acinetobacter baumannii bacteria. It focuses on the O-polysaccharides of A. baumannii and their role in the bacteria's virulence. The article presents a chemical synthesis strategy for the hexa- and pentasaccharide repeating units of the O-7 and O-10 antigens of A. baumannii. It describes the synthesis process, purification, and characterization of several compounds, providing valuable information for researchers studying these compounds. The article also mentions the availability of supporting information online. [Extracted from the article]

Phellinus igniarius (PI) has various kinds of biological activities, such as antitumour activities, and polysaccharides are one of its main components. In this study, polysaccharides from PI (PIP) were prepared, purified, analysed for their structure and investigated for their antitumour activity and mechanism in vitro. PIP consists of 12138 kDa of carbohydrates containing 90.5 ± 1.6% neutral carbohydrates. PIP consists of glucose, galactose, mannose, xylose, D-fructose, L-guluronic acid, glucosamine hydrochloride, rhamnose, arabinose and D-mannoturonic acid. PIP can significantly inhibit HepG2 cell proliferation, induce cell apoptosis and also inhibited migration and invasion in a concentration-dependent manner. PIP increased reactive oxygen species (ROS), increased the expression of p53, and induced cytochrome c release into the cytoplasm to activate caspase-3. PIP is a promising potential candidate for the therapeutic treatment of hepatic carcinoma via the ROS-mediated mitochondrial apoptosis pathway. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Komagataella phaffii (Pichia pastoris) has emerged as a common and robust biotechnological platform organism, to produce recombinant proteins and other bioproducts of commercial interest. Key advantage of K. phaffii is the secretion of recombinant proteins, coupled with a low host protein secretion. This facilitates downstream processing, resulting in high purity of the target protein. However, a significant but often overlooked aspect is the presence of an unknown polysaccharide impurity in the supernatant. Surprisingly, this impurity has received limited attention in the literature, and its presence and quantification are rarely addressed. Results: This study aims to quantify this exopolysaccharide in high cell density recombinant protein production processes and identify its origin. In stirred tank fed-batch fermentations with a maximal cell dry weight of 155 g/L, the polysaccharide concentration in the supernatant can reach up to 8.7 g/L. This level is similar to the achievable target protein concentration. Importantly, the results demonstrate that exopolysaccharide production is independent of the substrate and the protein production process itself. Instead, it is directly correlated with biomass formation and proportional to cell dry weight. Cell lysis can confidently be ruled out as the source of this exopolysaccharide in the culture medium. Furthermore, the polysaccharide secretion can be linked to a mutation in the HOC1 gene, featured by all derivatives of strain NRRL Y-11430, leading to a characteristic thinner cell wall. Conclusions: This research sheds light on a previously disregarded aspect of K. phaffii fermentations, emphasizing the importance of monitoring and addressing the exopolysaccharide impurity in biotechnological applications, independent of the recombinant protein produced. [ABSTRACT FROM AUTHOR]

Biofilm formation protects bacteria from antibiotic treatment and host immune responses, making biofilm infections difficult to treat. Within biofilms, bacterial cells are entangled in a self‐produced extracellular matrix that typically includes exopolysaccharides. Molecular‐level descriptions of biofilm matrix components, especially exopolysaccharides, have been challenging to attain due to their complex nature and lack of solubility and crystallinity. Solid‐state nuclear magnetic resonance (NMR) has emerged as a key tool to determine the structure of biofilm matrix exopolysaccharides without degradative sample preparation. In this review, we discuss challenges of studying biofilm matrix exopolysaccharides and opportunities to develop solid‐state NMR approaches to study these generally intractable materials. We specifically highlight investigations of the exopolysaccharide called Pel made by the opportunistic pathogen, Pseudomonas aeruginosa. We provide a roadmap for determining exopolysaccharide structure and discuss future opportunities to study such systems using solid‐state NMR. The strategies discussed for elucidating biofilm exopolysaccharide structure should be broadly applicable to studying the structures of other glycans. [ABSTRACT FROM AUTHOR]

Phytophthora capsici is an important plant pathogenic oomycete that causes great losses to vegetable production around the world. Antofine is an important alkaloid isolated from Cynanchum komarovii Al. Iljinski and exhibits significant antifungal activity. In this study, the effect of antofine on the mycelial growth, morphology, and physiological characteristics of P. capsici was investigated using colorimetry. Meanwhile, the activity of mitochondrial respiratory chain complexes of P. capsici was evaluated following treatment with a 30% effective concentration (EC30), as well as EC50 and EC70, of antofine for 0, 12, 24, and 48 h. The results showed that antofine had a significant inhibitory effect against P. capsici, with an EC50 of 5.0795 μg/mL. After treatment with antofine at EC50 and EC70, the mycelia were rough, less full, and had obvious depression; they had an irregular protrusion structure; and they had serious wrinkles. In P. capsici, oxalic acid and exopolysaccharide contents decreased significantly, while cell membrane permeability and glycerol content increased when treated with antofine. Reactive oxygen species (ROS) entered a burst state in P. capsici after incubation with antofine for 3 h, and fluorescence intensity was 2.43 times higher than that of the control. The activities of the mitochondrial respiratory chain complex II, III, I + III, II + III, V, and citrate synthase in P. capsici were significantly inhibited following treatment with antofine (EC50 and EC70) for 48 h compared to the control. This study revealed that antofine is likely to affect the pathways related to the energy metabolism of P. capsici and thus affect the activity of respiratory chain complexes. These results increase our understanding of the action mechanism of antofine against P. capsici. [ABSTRACT FROM AUTHOR]

Background and aims: The bicinchoninic acid (BCA) method was not yet applied on soil extracts of extracellular polymeric substances (EPS) to quantify polysaccharides, although this might be possible by introducing a cleavage step to produce monosaccharides. A pre-extraction with CaCl2 to remove interfering substances is usually performed before extracting EPS. The main objective of this study was to optimize the BCA assay for total carbohydrates quantification by applying a hydrolysis step to the EPS extracts, while also testing carbohydrate contents of CaCl2 pre-extracts. Methods: Total carbohydrates were quantified with BCA in EPS extracts of three soils, after hydrolysis with H2SO4, using two acid concentrations (0.75 and 1.0 M), three hydrolysis temperatures (100, 120 and 130 °C), and five hydrolysis times (10, 30, 50, 70, and 90 min). EPS were extracted with the cation exchange resin (CER) method adapted to soils. Two versions of pre-extraction with CaCl2 were tested twice consecutively. Results: More carbohydrates were measured after hydrolysis with 0.75 M H2SO4 at below 100 °C and after 10 min for all soils. Decreasing values were seen after longer reaction times and higher temperatures. CaCl2 extracted no or negligible amounts of carbohydrates from the soil. Conclusion: The pre-extraction step can be done without in most cases. The BCA assay is free of toxicity and easily performed, while also tolerant to interferences from most compounds in EPS extracts. These characteristics highlight the potential of this method for a rapid quantification of carbohydrates in studies of extractable polymers in several areas of soil biogeochemistry. [ABSTRACT FROM AUTHOR]

The bacterial genus Rhodococcus comprises organisms performing oleaginous behaviors under certain growth conditions and ratios of carbon and nitrogen availability. Rhodococci are outstanding producers of biofuel precursors, where lipid and glycogen metabolisms are closely related. Thus, a better understanding of rhodococcal carbon partitioning requires identifying catalytic steps redirecting sugar moieties to storage molecules. Here, we analyzed two GT4 glycosyl-transferases from Rhodococcus jostii (RjoGlgAb and RjoGlgAc) annotated as α-glucan-α-1,4-glucosyl transferases, putatively involved in glycogen synthesis. Both enzymes were produced in Escherichia coli cells, purified to homogeneity, and kinetically characterized. RjoGlgAb and RjoGlgAc presented the "canonical" glycogen synthase activity and were actives as maltose-1P synthases, although to a different extent. Then, RjoGlgAc is a homologous enzyme to the mycobacterial GlgM, with similar kinetic behavior and glucosyl-donor preference. RjoGlgAc was two orders of magnitude more efficient to glucosylate glucose-1P than glycogen, also using glucosamine-1P as a catalytically efficient aglycon. Instead, RjoGlgAb exhibited both activities with similar kinetic efficiency and preference for short-branched α-1,4-glucans. Curiously, RjoGlgAb presented a super-oligomeric conformation (higher than 15 subunits), representing a novel enzyme with a unique structure-to-function relationship. Kinetic results presented herein constitute a hint to infer on polysaccharides biosynthesis in rhodococci from an enzymological point of view. [ABSTRACT FROM AUTHOR]

A tomato‐infecting virus known as Beet curly top Iran virus (BCTIV) cause a significant disease for tomato plants and several other plant species around the world. Chitosan polysaccharide is a natural biopolymer that has been utilised as an exo‐elicitor to enhance plant defence mechanisms against a variety of plant diseases. This study investigates the efficacy of chitosan in combating BCTIV disease on tomato plants and modulating the host–virus interaction under greenhouse conditions. Twenty‐four hours before the virus inoculation, tomato plants were sprayed with a protective chitosan solution at different concentrations (0.5, 1, 1.5, and 2 mg/mL). Tomato plants were inoculated with a BCTIV infectious clone using an Agrobacterium‐inoculation method. The findings clearly demonstrated a reduction in the severity of the disease in chitosan‐treated plants as compared to Mock‐plants, with the percentage decreasing from 61.53% to 75.28% in 1.5 mg/mL treated plants and from 9.01% to 28.43% in 0.5 mg/mL treated plants. In addition, the utilisation of chitosan has the potential to deactivate the accumulation of BCTIV within the host tissues. The virus accumulation was greatly alleviated in 1, 1.5, and 2 mg/mL‐treated plants by 71.29%, 90.11%, and 93.14%, respectively, and over the mock plants. Furthermore, it was found that chitosan applied at all tested concentrations increased the relative expression and mRNA accumulation of genes related to resistance, including the pathogenesis‐related protein gene PR‐1, the HSP90 gene, and the AGO2a antiviral gene. These genes reached their maximum by 22.9‐, 12.93‐, and 4.44‐fold increases, respectively, over the untreated control. According to gas chromatography–mass spectroscop (GC‐MS) fractionation profile, chitosan increased 28 bioactive metabolic components, such as n‐hexadecanoic acid, heptanone, 1,2‐dimethylbenzene, dicarboxylic acid, and cis‐11‐octadecenoic acid methyl‐ester, to improve metabolic pathways. Results reported here revealed that foliar application of chitosan decreases the rate of the disease severity and virus accumulation in BCTIV‐infected tomato plants. This effect is associated with increased gene expression and defence‐related factors, enhancing tomato resistance to BCTIV infection. Consequently, chitosan treatments could be part of an integrated approach for reducing the severity of BCTIV disease in tomato and other host plants. [ABSTRACT FROM AUTHOR]

Bacillus thuringiensis (Bt) secretes the nutritional insecticidal protein Vip3Aa11, which exhibits high toxicity against the fall armyworm (Spodoptera frugiperda). The Bt HD270 extracellular polysaccharide (EPS) enhances the toxicity of Vip3Aa11 protoxin against S. frugiperda by enhancing the attachment of brush border membrane vesicles (BBMVs). However, how EPS-HD270 interacts with Vip3Aa11 protoxin in vivo and the effect of EPS-HD270 on the toxicity of activated Vip3Aa11 toxin are not yet clear. Our results indicated that there is an interaction between mannose, a monosaccharide that composes EPS-HD270, and Vip3Aa11 protoxin, with a dissociation constant of Kd = 16.75 ± 0.95 mmol/L. When EPS-HD270 and Vip3Aa11 protoxin were simultaneously fed to third-instar larvae, laser confocal microscopy observations revealed the co-localization of the two compounds near the midgut wall, which aggravated the damage to BBMVs. EPS-HD270 did not have a synergistic insecticidal effect on the activated Vip3Aa11 protein against S. frugiperda. The activated Vip3Aa11 toxin demonstrated a significantly reduced binding capacity (548.73 ± 82.87 nmol/L) towards EPS-HD270 in comparison to the protoxin (34.96 ± 9.00 nmol/L). Furthermore, this activation diminished the affinity of EPS-HD270 for BBMVs. This study provides important evidence for further elucidating the synergistic insecticidal mechanism between extracellular polysaccharides and Vip3Aa11 protein both in vivo and in vitro. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the immunomodulatory effect of the probiotic Limosilactobacillus reuteri L26 BiocenolTM (L26) and its purified exopolysaccharide (EPS) with respect to antiviral innate immune response. In our experiment, we used porcine epithelial IPEC-J2 cells as a model of the intestinal barrier in a homologous infection by porcine Rotavirus A strain OSU6 (RVA). The production of selected molecules of non-specific humoral immunity was evaluated at the mRNA level. The EPS alone significantly increased the level of interferon λ3 (IFN-λ3) mRNA in the non-infected IPEC-J2 cells (P < 0.001). We also tested whether the treatment of IPECJ2 cells by L26 or EPS influences the replication of RVA by virus titration and real-time PCR. We found that a pre-treatment in combination with subsequent continuous stimulation has no influence on the RVA replication. However, both treatments significantly decreased the RVA-induced production of IFN-λ3 (P < 0.05) and the "SOS" cytokine interleukin 6 (IL-6; P < 0.01), already at the transcription level. In addition, the EPS treatment resulted in significantly increased IL-10 mRNA in the RVA-infected cells. In summary, we assume an immunoregulatory potential of L. reuteri L26 BiocenolTM and its EPS in the local intestinal antiviral immune response. [ABSTRACT FROM AUTHOR]

The simulated digestion and fermentation characteristics in vitro of exopolysaccharide (EPS) of Levilactobacillus brevis M‐10 were studied to evaluate its postbiotic properties. The simulated digestion results showed that EPS could not be degraded in saliva but could be very slightly degraded in gastric juice and could be degraded in intestinal juice. The results of simulated fermentation demonstrated that EPS could lower the intestine pH and be utilized by gut microbes to produce short‐chain fatty acids such as propionic acid and butyric acid. Meanwhile, EPS significantly raised the diversity of human gut microbiota, and the relative abundances of Phascolarctobacterium were significantly increased, whereas Fusobacterium and Morganella significantly decreased. In conclusion, EPS from L. brevis M‐10 was a good postbiotic as inulin. This was the first report about EPS as the postbiotic of L. brevis M‐10 screened from broomcorn millet sour porridge in northwestern Shanxi Province, China. [ABSTRACT FROM AUTHOR]

The unique pectin-like carbohydrate "sphagnan" has been shown to protect organic matter from microbial decomposition in Sphagnum-dominated peatlands. However, the bioavailability of sphagnan has not been evaluated, and it is not known if it persists or continues to affect decomposition processes over the long timescales of peat formation. To address this, we assessed the connection between sphagnan content and organic matter decomposition rates in a temperate peatland near Fennville, MI, USA. We compared the effects of sphagnan over two timescales: (1) a short-term litter incubation assay using mosses from different peatland microtopographies; and (2) oxic and anoxic incubation assays of peat collected from multiple depths within the peat profile, reflecting a natural long-term decomposition continuum. On both timescales, we hypothesized that higher sphagnan content would be associated with lower decomposition rates, and that sphagnan would be selectively preserved compared to bulk C and other carbohydrates. The litter decomposition experiment supported both hypotheses, as higher sphagnan content was associated with lower mass loss, and sphagnan content increased due to selective preservation. In the peat, we observed weak but significant correlations between the relative abundance of sphagnan (as a fraction of total non-cellulosic sugars) and both aerobic and anaerobic respiration rates. This relationship was stronger in cores collected from hollow microtopographies than those from hummocks. However, there was not a significant relationship between respiration rates and the total (C-normalized) sphagnan content. Sphagnan content increased with depth in the peat profile, indicating selective preservation compared to bulk C. Additionally, we observed the accumulation of non-cellulosic glucose in the deep peat, likely derived from microbial exopolysaccharides. Together, these results indicate that sphagnan persists in the catotelm and continues to contribute to the long-term stabilization of organic matter in Sphagnum-rich peatlands, although the weak relationship with respiration indicates that its influence is relatively minor. [ABSTRACT FROM AUTHOR]

Background: The current study was carried out to assess the alternative role of high temperature tolerant plant growth promoting rhizobacteria (PGPR) in alleviation of heat stress adverse effect in eggplant. Methods: PGPR was assessed for PGP potentialities and investigated for1-aminocyclopropane-1-carboxylate (ACC deaminase), and exopolysaccharide under normal and high temperature environment. The impact of PGPR application on plant physiology and biochemistry was investigated under normal and high temperature environments and plant growth regulators were evaluated using High Performance Liquid Chromatography (HPLC). Results: A significant impediment for morphological and physiological parameters of eggplant cultivars was observed during heat stress in absence of PGPR inoculation. The results showed that ACC-deaminase produced by PGPR boosted up significantly the extracellular polymeric substances (EPS) accumulation, conversion of ACC into a-ketobutyrate and ammonia, and reduce consequently the impact of high temperature on eggplant development. Conclusion: PGPR inoculation showed an alternative strategy to improve eggplant growth and development under high temperature condition in order to preserve agriculture sustainability and healthy crops. [ABSTRACT FROM AUTHOR]