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1.
Curr Genet ; 70(1): 7, 2024 May 14.
Article in English | MEDLINE | ID: mdl-38743270

ABSTRACT

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


Subject(s)
Fermentation , Lacticaseibacillus paracasei , Oxidative Stress , Polysaccharides, Bacterial , Solanum melongena , Polysaccharides, Bacterial/biosynthesis , Polysaccharides, Bacterial/metabolism , Solanum melongena/microbiology , Solanum melongena/genetics , Solanum melongena/metabolism , Lacticaseibacillus paracasei/metabolism , Lacticaseibacillus paracasei/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Antioxidants/metabolism , Hydrogen Peroxide/metabolism , Genome, Bacterial , Fermented Foods/microbiology , Superoxide Dismutase/metabolism , Superoxide Dismutase/genetics
2.
BMC Plant Biol ; 24(1): 450, 2024 May 23.
Article in English | MEDLINE | ID: mdl-38783216

ABSTRACT

BACKGROUND: Guava is a fruit prone to rapid spoilage following harvest, attributed to continuous and swift physicochemical transformations, leading to substantial postharvest losses. This study explored the efficacy of xanthan gum (XG) coatings applied at various concentrations (0.25, 0.5, and 0.75%) on guava fruits (Gola cultivar) over a 15-day storage period. RESULTS: The results indicated that XG coatings, particularly at 0.75%, substantially mitigated moisture loss and decay, presenting an optimal concentration. The coated fruits exhibited a modified total soluble soluble solids, an increased total titratable acidity, and an enhanced sugar-acid ratio, collectively enhancing overall quality. Furthermore, the XG coatings demonstrated the remarkable ability to preserve bioactive compounds, such as total phenolics, flavonoids, and antioxidants, while minimizing the levels of oxidative stress markers, such as electrolyte leakage, malondialdehyde, and H2O2. The coatings also influenced cell wall components, maintaining levels of hemicellulose, cellulose, and protopectin while reducing water-soluble pectin. Quantitative analysis of ROS-scavenging enzymes, including superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase, revealed significant increases in their activities in the XG-coated fruits compared to those in the control fruits. Specifically, on day 15, the 0.75% XG coating demonstrated the highest SOD and CAT activities while minimizing the reduction in APX activity. Moreover, XG coatings mitigated the activities of fruit-softening enzymes, including pectin methylesterase, polygalacturonase, and cellulase. CONCLUSIONS: This study concludes that XG coatings play a crucial role in preserving postharvest quality of guava fruits by regulating various physiological and biochemical processes. These findings offer valuable insights into the potential application of XG as a natural coating to extend the shelf life and maintain the quality of guava fruits during storage.


Subject(s)
Fruit , Polysaccharides, Bacterial , Psidium , Psidium/chemistry , Polysaccharides, Bacterial/pharmacology , Fruit/chemistry , Fruit/drug effects , Food Preservation/methods , Antioxidants/metabolism
3.
Int J Biol Macromol ; 268(Pt 2): 131975, 2024 May.
Article in English | MEDLINE | ID: mdl-38692551

ABSTRACT

Vitamin E (VE) microencapsulation using a green surfactant emulsifier not only protects the active substance and is also environmentally friendly. In this study, we used alcohol ether glycoside as an emulsifier to prepare VE microcapsules using the biological macromolecule Zein and various polysaccharides. The resulting nano microcapsules exhibited a spherical structure, stable morphology, uniform size, and a >90% encapsulation efficiency. They also had good thermal stability and slow-release properties. Of these, xanthan gum/Zein-VE microcapsules were superior, with antioxidant properties up to 3.05-fold higher than untreated VE. We successfully developed VE nano microcapsules that meet eco-friendly and sustainable requirements, which may have applications in the food and pharmaceutical industries.


Subject(s)
Antioxidants , Capsules , Polysaccharides , Vitamin E , Zein , Zein/chemistry , Vitamin E/chemistry , Polysaccharides/chemistry , Antioxidants/chemistry , Antioxidants/pharmacology , Polysaccharides, Bacterial/chemistry , Particle Size , Drug Compounding/methods
4.
J Bacteriol ; 206(5): e0004824, 2024 May 23.
Article in English | MEDLINE | ID: mdl-38712944

ABSTRACT

Whole genome sequencing has revealed that the genome of Staphylococcus aureus possesses an uncharacterized 5-gene operon (SAOUHSC_00088-00092 in strain 8325 genome) that encodes factors with functions related to polysaccharide biosynthesis and export, indicating the existence of a new extracellular polysaccharide species. We designate this locus as ssc for staphylococcal surface carbohydrate. We found that the ssc genes were weakly expressed and highly repressed by the global regulator MgrA. To characterize Ssc, Ssc was heterologously expressed in Escherichia coli and extracted by heat treatment. Ssc was also conjugated to AcrA from Campylobacter jejuni in E. coli using protein glycan coupling technology (PGCT). Analysis of the heat-extracted Ssc and the purified Ssc-AcrA glycoconjugate by tandem mass spectrometry revealed that Ssc is likely a polymer consisting of N-acetylgalactosamine. We further demonstrated that the expression of the ssc genes in S. aureus affected phage adsorption and susceptibility, suggesting that Ssc is surface-exposed. IMPORTANCE: Surface polysaccharides play crucial roles in the biology and virulence of bacterial pathogens. Staphylococcus aureus produces four major types of polysaccharides that have been well-characterized. In this study, we identified a new surface polysaccharide containing N-acetylgalactosamine (GalNAc). This marks the first report of GalNAc-containing polysaccharide in S. aureus. Our discovery lays the groundwork for further investigations into the chemical structure, surface location, and role in pathogenesis of this new polysaccharide.


Subject(s)
Acetylgalactosamine , Polysaccharides, Bacterial , Staphylococcus aureus , Staphylococcus aureus/genetics , Staphylococcus aureus/metabolism , Acetylgalactosamine/metabolism , Polysaccharides, Bacterial/metabolism , Polysaccharides, Bacterial/genetics , Polysaccharides, Bacterial/chemistry , Gene Expression Regulation, Bacterial , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Campylobacter jejuni/genetics , Campylobacter jejuni/metabolism
5.
Toxins (Basel) ; 16(5)2024 May 07.
Article in English | MEDLINE | ID: mdl-38787067

ABSTRACT

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.


Subject(s)
Bacterial Proteins , Polysaccharides, Bacterial , Spodoptera , Animals , Bacterial Proteins/toxicity , Bacterial Proteins/metabolism , Polysaccharides, Bacterial/pharmacology , Polysaccharides, Bacterial/chemistry , Spodoptera/drug effects , Larva/drug effects , Insecticides/toxicity , Insecticides/pharmacology , Bacillus thuringiensis/metabolism , Microvilli/metabolism , Microvilli/drug effects
6.
Int J Mol Sci ; 25(9)2024 May 03.
Article in English | MEDLINE | ID: mdl-38732217

ABSTRACT

The Euganean Thermal District, situated in North-East Italy, is one of Europe's largest and oldest thermal centres. The topical application of its therapeutic thermal muds is recognised by the Italian Health System as a beneficial treatment for patients suffering from arthro-rheumatic diseases. Polysaccharides produced by the mud microbiota have been recently identified as anti-inflammatory bioactive molecules. In this paper we analysed the efficacy of Microbial-Polysaccharides (M-PS) derived from mature muds obtained at different maturation temperatures, both within and outside the codified traditional mud maturation range. M-PSs were extracted from six mature muds produced by five spas of the Euganean Thermal District and investigated for their chemical properties, monosaccharide composition and in vivo anti-inflammatory potential, using the zebrafish model organism. Additionally, mature muds were characterized for their microbiota composition using Next-Generation Sequencing. The results showed that all M-PSs exhibit similar anti-inflammatory potential, referable to their comparable chemical composition. This consistency was observed despite changes in cyanobacteria populations, suggesting a possible role of the entire microbial community in shaping the properties of these biomolecules. These findings highlight the importance of scientific research in untangling the origins of the therapeutic efficacy of Euganean Thermal muds in the treatment of chronic inflammatory conditions.


Subject(s)
Anti-Inflammatory Agents , Zebrafish , Animals , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/chemistry , Italy , Polysaccharides, Bacterial/pharmacology , Polysaccharides, Bacterial/chemistry , Microbiota/drug effects , Polysaccharides/pharmacology , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Mud Therapy
7.
Int J Immunopathol Pharmacol ; 38: 3946320241249429, 2024.
Article in English | MEDLINE | ID: mdl-38721971

ABSTRACT

OBJECTIVE: This study investigated the raft-forming suspension of famotidine as an anti-reflux formulation to improve the oral bioavailability of narrow absorption window drugs by enhancing gastric residence time (GRT) and preventing gastro-esophageal reflux disease (GERD). METHOD: Various combinations of raft-forming agents, such as Tragacanth gum (TG), guar gum (GG), and xanthan gum (XG), were evaluated alongside sodium alginate (SA) to develop an effective raft. Preformulation studies and preliminary screening were conducted to identify the most suitable raft-forming agent, and GG was chosen due to its mucilaginous properties. The formulation was optimized using a 32 full factorial design, with the quantities of GG and SA as independent factors and apparent viscosity and in-vitro drug release (%) as dependent factors. The in vivo floating behavior study was performed for optimized and stabilized formulation. RESULTS: Among the tested batches, F6 was selected as the optimized formulation. It exhibited desirable characteristics such as adequate raft weight for extended floating in gastric fluid, improved apparent viscosity, and a significant percentage of drug release at 12 h. A mathematical model was applied to the in-vitro data to gain insights into the drug release mechanism of the formulation. The stability of the suspension was assessed under accelerated conditions, and it demonstrated satisfactory stability. The formulation remains floating in the Rabbit stomach for more than 12 h. CONCLUSION: It concludes that the developed formulation has enhanced bioavailability in the combination of GG and SA. The floating layer of the raft prevents acid reflux, and the famotidine is retained for an extended period of time in the gastric region, preventing excess acid secretion. The developed formulations are effective for stomach ulcers and GERD, with the effect of reducing acid secretion by H2 receptor antagonists.


Subject(s)
Drug Delivery Systems , Famotidine , Galactans , Famotidine/administration & dosage , Famotidine/pharmacokinetics , Animals , Drug Delivery Systems/methods , Drug Liberation , Alginates , Gastroesophageal Reflux/drug therapy , Gastroesophageal Reflux/metabolism , Biological Availability , Mannans/administration & dosage , Plant Gums , Viscosity , Male , Rabbits , Gastric Mucosa/metabolism , Gastric Mucosa/drug effects , Polysaccharides, Bacterial , Drug Stability , Administration, Oral
8.
J Microbiol Methods ; 221: 106942, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38704038

ABSTRACT

Methylation analysis was performed on methylated alditol acetate standards and Streptococcus mutans extracellular polymeric substances (EPS) produced from wild-type and Gtf knockout strains (∆GtfB, ∆GtfB, and ∆GtfD). The methylated alditol acetate standards were representative of glycosidic linkages found in S. mutans EPS and were used to calibrate the GC-MS system for an FID detector and MS (TIC) and produce molar response factor, a necessary step in quantitative analysis. FID response factors were consistent with literature values (Sweet et al., 1975) and found to be the superior option for quantitative results, although the TIC response factors now give researchers without access to an FID detector a needed option for molar response factor correction. The GC-MS analysis is then used to deliver the ratio of the linkage types within a biofilm.


Subject(s)
Biofilms , Gas Chromatography-Mass Spectrometry , Polysaccharides, Bacterial , Streptococcus mutans , Biofilms/growth & development , Streptococcus mutans/genetics , Streptococcus mutans/metabolism , Gas Chromatography-Mass Spectrometry/methods , Polysaccharides, Bacterial/metabolism , Glycosides/metabolism , Methylation , Extracellular Polymeric Substance Matrix/metabolism , Extracellular Polymeric Substance Matrix/chemistry , Polysaccharides/metabolism
9.
Nat Commun ; 15(1): 4048, 2024 May 14.
Article in English | MEDLINE | ID: mdl-38744821

ABSTRACT

Phytoplankton blooms provoke bacterioplankton blooms, from which bacterial biomass (necromass) is released via increased zooplankton grazing and viral lysis. While bacterial consumption of algal biomass during blooms is well-studied, little is known about the concurrent recycling of these substantial amounts of bacterial necromass. We demonstrate that bacterial biomass, such as bacterial alpha-glucan storage polysaccharides, generated from the consumption of algal organic matter, is reused and thus itself a major bacterial carbon source in vitro and during a diatom-dominated bloom. We highlight conserved enzymes and binding proteins of dominant bloom-responder clades that are presumably involved in the recycling of bacterial alpha-glucan by members of the bacterial community. We furthermore demonstrate that the corresponding protein machineries can be specifically induced by extracted alpha-glucan-rich bacterial polysaccharide extracts. This recycling of bacterial necromass likely constitutes a large-scale intra-population energy conservation mechanism that keeps substantial amounts of carbon in a dedicated part of the microbial loop.


Subject(s)
Bacteria , Carbon Cycle , Glucans , Glucans/metabolism , Bacteria/metabolism , Bacteria/classification , Bacteria/genetics , Phytoplankton/metabolism , Biomass , Diatoms/metabolism , Eutrophication , Carbon/metabolism , Zooplankton/metabolism , Polysaccharides, Bacterial/metabolism , Polysaccharides, Bacterial/chemistry , Bacterial Proteins/metabolism
10.
Sci Rep ; 14(1): 11011, 2024 05 14.
Article in English | MEDLINE | ID: mdl-38744937

ABSTRACT

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.


Subject(s)
Silk , Spiders , Animals , Spiders/microbiology , Spiders/metabolism , Silk/metabolism , Bacteria/metabolism , Polysaccharides, Bacterial/metabolism
11.
Proc Natl Acad Sci U S A ; 121(21): e2402554121, 2024 May 21.
Article in English | MEDLINE | ID: mdl-38748580

ABSTRACT

Cell surface glycans are major drivers of antigenic diversity in bacteria. The biochemistry and molecular biology underpinning their synthesis are important in understanding host-pathogen interactions and for vaccine development with emerging chemoenzymatic and glycoengineering approaches. Structural diversity in glycostructures arises from the action of glycosyltransferases (GTs) that use an immense catalog of activated sugar donors to build the repeating unit and modifying enzymes that add further heterogeneity. Classical Leloir GTs incorporate α- or ß-linked sugars by inverting or retaining mechanisms, depending on the nucleotide sugar donor. In contrast, the mechanism of known ribofuranosyltransferases is confined to ß-linkages, so the existence of α-linked ribofuranose in some glycans dictates an alternative strategy. Here, we use Citrobacter youngae O1 and O2 lipopolysaccharide O antigens as prototypes to describe a widespread, versatile pathway for incorporating side-chain α-linked pentofuranoses by extracytoplasmic postpolymerization glycosylation. The pathway requires a polyprenyl phosphoribose synthase to generate a lipid-linked donor, a MATE-family flippase to transport the donor to the periplasm, and a GT-C type GT (founding the GT136 family) that performs the final glycosylation reaction. The characterized system shares similarities, but also fundamental differences, with both cell wall arabinan biosynthesis in mycobacteria, and periplasmic glucosylation of O antigens first discovered in Salmonella and Shigella. The participation of auxiliary epimerases allows the diversification of incorporated pentofuranoses. The results offer insight into a broad concept in microbial glycobiology and provide prototype systems and bioinformatic guides that facilitate discovery of further examples from diverse species, some in currently unknown glycans.


Subject(s)
Glycosyltransferases , Glycosyltransferases/metabolism , Glycosyltransferases/genetics , Glycosylation , Citrobacter/metabolism , Citrobacter/genetics , O Antigens/metabolism , O Antigens/chemistry , Polysaccharides/metabolism , Bacterial Proteins/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/chemistry , Polysaccharides, Bacterial/metabolism
12.
Int J Pharm Compd ; 28(3): 249-259, 2024.
Article in English | MEDLINE | ID: mdl-38768505

ABSTRACT

Since ancient times, mouth fresheners in many different forms have been used throughout the world. Traditional knowledge describes the health benefits of mouth fresheners, and contemporary science is now investigating their benefits. Claims have been made that mouth fresheners not only improve digestion but also promote oral health. Similar, but in a more profound sense, probiotics offer astounding advantages in treating many disorders. In certain cases, probiotics also offer prophylactic effects. Numerous benefits for dental health are being studied for B. coagulans (MB-BCM9) and B. subtilis (MB-BSM12). In this current study, a probiotic and a mouth freshener were combined to ameliorate the impacts of both. The oral residence of probiotics was enhanced by employing mucoadhesive polymers. Numerous compositions were developed and evaluated for the unaltered growth of probiotics, along with other evaluations like microscopy, in vitro mucoadhesive strength, and stability studies. Xanthan gum and hydroxypropyl methylcellulose were used in the development of mucoadhesive probiotic powder by employing the lyophilization technique. More than five hours of residence time were observed in the in vitro study with goat oral mucosa. The enumeration study validated the label claims of MB-BCM9 and MB-BSM12. It also concluded that none of the components of the formulation had a detrimental effect on probiotics. In essence, the present work discloses the novel and stable formulation of a probiotic-based mouth freshener.


Subject(s)
Hypromellose Derivatives , Mouth Mucosa , Polysaccharides, Bacterial , Probiotics , Probiotics/administration & dosage , Animals , Hypromellose Derivatives/chemistry , Polysaccharides, Bacterial/chemistry , Goats , Adhesiveness , Freeze Drying , Drug Compounding , Powders , Drug Stability
13.
Food Res Int ; 187: 114329, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38763632

ABSTRACT

The utilization of non-animal-derived materials to imitate cartilage is critical for the advancement of plant-based simulated meat. In this study, gellan gum (GG), konjac glucomannan (KGM), and wheat fiber (WF) were used to construct hydrogel, and the mechanical strength, water properties, and microstructure were regulated by constructing Ca2+ cross-links and moisture control. The hardness, chewiness, resilience, shear force, and shear energy of the Ca2+ cross-linked samples were significantly improved. Extrusion dehydration further changes the related mechanical properties of the hydrogel and results in a tighter microstructure. The findings suggest that the establishment of Ca2+ cross-links and water regulation are efficacious techniques for modifying the texture of the GG/KGM/WF composite hydrogel. Correlation analysis and sensory evaluation showed that the test indexes and sensory scores of the samples with Ca2+ crosslinking and 80 % moisture content were similar to chicken breast cartilage, and the samples with Ca2+ crosslinking and 70 % moisture content were similar to pig crescent bone. This study presents a framework for designing edible cartilage simulators using polysaccharide hydrogels, with implications for enhancing the resemblance of plant-based meat products to real meat and expanding the range of vegetarian offerings available.


Subject(s)
Hydrogels , Mannans , Polysaccharides, Bacterial , Triticum , Polysaccharides, Bacterial/chemistry , Mannans/chemistry , Animals , Hydrogels/chemistry , Triticum/chemistry , Cartilage/chemistry , Water/chemistry , Cross-Linking Reagents/chemistry , Chickens , Calcium/analysis , Calcium/chemistry , Dietary Fiber/analysis
14.
Pediatr Infect Dis J ; 43(2): e67-e70, 2024 Feb 01.
Article in English | MEDLINE | ID: mdl-38758207

ABSTRACT

We evaluated whether the quantification of IgG to pneumococcal capsular polysaccharides is an accurate diagnostic test for pneumococcal infection in children with pneumonia in Nepal. Children with pneumococcal pneumonia did not have higher convalescent, or higher fold change, IgG to pneumococcal polysaccharides than children with other causes of pneumonia. Caution is needed in interpreting antibody responses in pneumococcal infections.


Subject(s)
Antibodies, Bacterial , Community-Acquired Infections , Immunoglobulin G , Pneumonia, Pneumococcal , Polysaccharides, Bacterial , Streptococcus pneumoniae , Humans , Antibodies, Bacterial/blood , Child, Preschool , Polysaccharides, Bacterial/immunology , Immunoglobulin G/blood , Infant , Streptococcus pneumoniae/immunology , Pneumonia, Pneumococcal/diagnosis , Pneumonia, Pneumococcal/immunology , Community-Acquired Infections/diagnosis , Community-Acquired Infections/immunology , Male , Female , Child , Nepal , Bacterial Capsules/immunology
15.
Arch Oral Biol ; 163: 105976, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38640776

ABSTRACT

OBJECTIVE: The present study investigated the effects of 4-hydroxy-3-methoxybenzaldehyde (4-H-3-MB) against Streptococcus mutans (S. mutans) using an in vitro cariogenic biofilm model. DESIGN: The antimicrobial susceptibility of biofilm-forming S. mutans was evaluated by disc diffusion method. In vitro investigations were performed using crystal violet staining assay (biofilm assay), exopolysaccharide (EPS) assay, acid production, growth curve analysis, optical microscopic, and FE-SEM analyses to determine the antibiofilm activity of 4-H-3-MB. RESULTS: S. mutans (SDC-05) was resistant to ampicillin, piperacillin/tazobactam and ceftriaxone, whereas the other strains of S. mutans (SDC-01, 02, 03 and SDC-04) were sensitive to all the antibiotics tested. 4-H-3-MB showed promising antibiofilm activity on S. mutans UA159 (79.81 %, 67.76 % and 56.31 %) and S. mutans SDC-05 (77.00 %, 59.48 % and 48.22 %) at the lowest concentration of 0.2, 0.1, 0.05 mg/ml. 4-H-3-MB did not inhibit bacterial growth even at concentrations 0.2 mg/ml. Similarly, 4-H-3-MB led to significant attrition in exopolysaccharide (EPS) and acid production by S. mutans UA159 and S. mutans (SDC-05) at the concentration of 0.2, 0.1 mg/ml, respectively. Optical microscopy and FE-SEM analysis 4-H-3-MB reduced the biofilm thickness of S. mutans UA159 and S. mutans SDC-05 relative to the untreated specimens. CONCLUSION: 4-H-3-MB significantly inhibited biofilm formation by S. mutans in a dose-dependent manner. Hence, our findings indicate that the active principle of 4-H-3-MB could be used as a biofilm inhibiting agent against S. mutans.


Subject(s)
Anti-Bacterial Agents , Benzaldehydes , Biofilms , Microbial Sensitivity Tests , Quorum Sensing , Streptococcus mutans , Virulence Factors , Streptococcus mutans/drug effects , Biofilms/drug effects , Quorum Sensing/drug effects , Benzaldehydes/pharmacology , Anti-Bacterial Agents/pharmacology , Polysaccharides, Bacterial/pharmacology , Microscopy, Electron, Scanning , In Vitro Techniques
16.
Appl Environ Microbiol ; 90(5): e0033424, 2024 May 21.
Article in English | MEDLINE | ID: mdl-38624197

ABSTRACT

Aggregating strains of Tetragenococcus halophilus tend to be trapped during soy sauce mash-pressing process and are, therefore, critical for clear soy sauce production. However, the precise molecular mechanism involved in T. halophilus aggregation remains elusive. In previous studies, we isolated a number of aggregating strains, including T. halophilus AB4 and AL1, and showed that a cell surface proteinaceous aggregation factor is responsible for their aggregation phenotype. In the present study, we explored the role of polysaccharide intercellular adhesin (PIA) in aggregate formation in T. halophilus SL10, isolated from soy sauce. SL10 exhibited similar aggregation to AB4 and AL1 but formed a non-uniform precipitate with distinctive wrinkles at the bottom of the test tube, unlike AB4 and AL1. Insertion sequence mutations in each gene of the ica operon diminished aggregation and PIA production, highlighting the critical role of IcaADBC-mediated PIA production in T. halophilus aggregation. Furthermore, two non-aggregating cardiolipin synthase (cls) gene mutants with intact ica operon did not produce detectable PIA. Phospholipid composition analysis in cls mutants revealed a decrease in cardiolipin and an increase in phosphatidylglycerol levels, highlighting the association between phospholipid composition and PIA production. These findings provide evidence for the pivotal role of cls in PIA-mediated aggregation and lay the foundation for future studies to understand the intricate networks of the multiple aggregation factors governing microbial aggregation.IMPORTANCEAggregation, commonly observed in various microbes, triggers biofilm formation in pathogenic variants and plays a beneficial role in efficient food production in those used for food production. Here, we showed that Tetragenococcus halophilus, a microorganism used in soy sauce fermentation, forms aggregates in a polysaccharide intercellular adhesin (PIA)-mediated manner. Additionally, we unveiled the relationship between phospholipid composition and PIA production. This study provides evidence for the presence of aggregation factors in T. halophilus other than the proteinaceous aggregation factor and suggests that further understanding of the coordinated action of these factors may improve clarified soy sauce production.


Subject(s)
Phospholipids , Phospholipids/metabolism , Enterococcaceae/metabolism , Enterococcaceae/genetics , Polysaccharides, Bacterial/metabolism , Bacterial Adhesion , Bacterial Proteins/metabolism , Bacterial Proteins/genetics
17.
Microb Pathog ; 191: 106664, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38679245

ABSTRACT

Pseudomonas aeruginosa causes life-threatening diseases and is resistant to almost all conventional antibiotics. The quorum sensing (QS) system of P. aeruginosa contributes to many pathogenic factors some of which are pigment production, motility, and biofilm. The disruption of quorum sensing system may be an impactful strategy to deal with infections. The present study investigates the anti-quorum sensing property of a bioactive molecule extracted from marine epibiotic bacteria present on the surface of seaweeds. Among all the isolates tested against monitor strain Chromobacterium violaceum (MTCC 2656), the one with the highest activity was identified as Bacillus zhangzhouensis SK4. The culture supernatant was extracted with chloroform which was then partially purified by TLC and column chromatography. The probable anti-QS compound was identified as 1,2-benzenedicarboxylic acid, bis (2-methylpropyl ester) by GC-MS and NMR analysis. The treatment of P. aeruginosa MCC 3457 with the lead compound resulted in the reduced production of pyocyanin, rhamnolipids, exopolysaccharide, biofilm, and motility. The observations of light and scanning electron microscopy also supported the biofilm inhibition. The lead compound showed synergism with the meropenem antibiotic and significantly reduced MIC. The molecular docking and pharmacokinetics study predicted 1, 2-benzenedicarboxylic acid, bis (2-methylpropyl ester), a phthalate derivative as a good drug candidate. The molecular dynamics study was also performed to check the stability of the lead compound and LasR complex. Further, lead compounds did not exhibit any cytotoxicity when tested on human embryonic kidney cells. As per our knowledge, this is the first report on the anti-QS activity of B. zhangzhouensis SK4, indicating that epibiotic bacteria can be a possible source of novel compounds to deal with the multidrug resistance phenomenon.


Subject(s)
Anti-Bacterial Agents , Bacillus , Biofilms , Molecular Docking Simulation , Pseudomonas aeruginosa , Quorum Sensing , Virulence Factors , Quorum Sensing/drug effects , Pseudomonas aeruginosa/drug effects , Biofilms/drug effects , Biofilms/growth & development , Virulence Factors/metabolism , Humans , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Bacillus/drug effects , Bacillus/chemistry , Bacillus/metabolism , Chromobacterium/drug effects , Microbial Sensitivity Tests , Pyocyanine/metabolism , Bacterial Proteins/metabolism , Glycolipids/pharmacology , Glycolipids/chemistry , Polysaccharides, Bacterial/pharmacology , Polysaccharides, Bacterial/isolation & purification , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/metabolism
18.
Int J Biol Macromol ; 267(Pt 1): 131166, 2024 May.
Article in English | MEDLINE | ID: mdl-38582464

ABSTRACT

Here, the simultaneous effect of chemo- and photothermal therapy against epidermoid carcinoma (EC) was investigated. A novel hydrogel, termed bionanogel (BNG), was designed using psyllium mucilage polysaccharide and bacterial gellan gum, incorporated with nanocomplex carrying caffeic acid (CA) and IR-820, and further characterized. The dual effect of BNG and 808 nm laser (BNG + L) on EC was investigated. Staining and scratch assays were performed to analyze their therapeutic effect on EC. In vivo evaluations of BNG + L in xenograft models were performed. Rapid transition, limited swelling, degradability and high tensile strength indicated BNG stability and sustained drug release. Irradiation with 808 nm laser light at 1.25 W /cm2 for 4 min resulted in a temperature increase of 53 °C and facilitated cell ablation. The in vitro studies showed that BNG + L suppressed cancer progression via a late apoptotic effect. The in vivo study showed that the slow release of CA from BNG + L significantly attenuated EC with low mitotic index and downregulation of proteins involved in cancer proliferation such as EGFR, AKT, PI3K, ERK, mTOR and HIF-1α. Thus, BNG could be a novel medium for targeted and controlled drug delivery for the treatment of epidermoid cancer when triggered by NIR light.


Subject(s)
Caffeic Acids , Carcinoma, Squamous Cell , Polysaccharides, Bacterial , Psyllium , Caffeic Acids/pharmacology , Caffeic Acids/chemistry , Caffeic Acids/administration & dosage , Animals , Humans , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/pharmacology , Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/pathology , Mice , Psyllium/chemistry , Psyllium/pharmacology , Cell Line, Tumor , Polysaccharides/chemistry , Polysaccharides/pharmacology , Hydrogels/chemistry , Xenograft Model Antitumor Assays , Drug Delivery Systems
19.
Int J Biol Macromol ; 267(Pt 1): 131377, 2024 May.
Article in English | MEDLINE | ID: mdl-38583850

ABSTRACT

Kombucha is prepared by fermenting sugared green or black tea with a symbiotic culture of bacteria and yeast (SCOBY). Some of the bacteria within the SCOBY are known to form exopolysaccharides (EPS) from sucrose. However, it is yet unknown whether water-soluble EPS are formed in kombucha, and if so, which specific EPS are present. Therefore, different kombucha samples were prepared by fermentation of green and black tea with SCOBYs from different manufacturers. Subsequently, the EPS were isolated and characterized by using various chromatographic methods, partial enzymatic hydrolyses and NMR spectroscopy. It was demonstrated that levans with a varying degree of branching at position O1 (4.3-7.9 %) are present, while only trace amounts of glucans were detected. Furthermore, levans isolated from kombucha had a comparably low molecular weight and the content of levan within the kombucha samples varied from 33 to 562 mg levan/L kombucha. Therefore, our study demonstrated that levans are the main EPS type in kombucha and that levan amounts and structures varied when different starter cultures and ingredients were used. Furthermore, we provide a comprehensive data set on the structural variability of levans from kombucha.


Subject(s)
Fermentation , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/isolation & purification , Molecular Weight , Kombucha Tea/microbiology , Fructans/chemistry , Fructans/isolation & purification , Magnetic Resonance Spectroscopy
20.
Int J Biol Macromol ; 267(Pt 2): 131521, 2024 May.
Article in English | MEDLINE | ID: mdl-38608976

ABSTRACT

Herein, the effects of anionic xanthan gum (XG), neutral guar gum (GG), and neutral konjac glucomannan (KGM) on the dissolution, physicochemical properties, and emulsion stabilization ability of soy protein isolate (SPI)-polysaccharide conjugates were studied. The SPI-polysaccharide conjugates had better water dissolution than the insoluble SPI. Compared with SPI, SPI-polysaccharide conjugates had lower ß-sheet (39.6 %-56.4 % vs. 47.3 %) and α-helix (13.0 %-13.2 % vs. 22.6 %) percentages, and higher ß-turn (23.8 %-26.5 % vs. 11.0 %) percentages. The creaming stability of SPI-polysaccharide conjugate-stabilized fish oil-loaded emulsions mainly depended on polysaccharide type: SPI-XG (Creaming index: 0) > SPI-GG (Creaming index: 8.1 %-21.2 %) > SPI-KGM (18.1 %-40.4 %). In addition, it also depended on the SPI preparation concentrations, glycation times, and glycation pH. The modification by anionic XG induced no obvious emulsion creaming even after 14-day storage, which suggested that anionic polysaccharide might be the best polysaccharide to modify SPI for emulsion stabilization. This work provided useful information to modify insoluble proteins by polysaccharides for potential application.


Subject(s)
Emulsions , Fish Oils , Galactans , Mannans , Plant Gums , Polysaccharides, Bacterial , Solubility , Soybean Proteins , Mannans/chemistry , Polysaccharides, Bacterial/chemistry , Plant Gums/chemistry , Emulsions/chemistry , Soybean Proteins/chemistry , Galactans/chemistry , Fish Oils/chemistry , Anions/chemistry
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