Polysaccharide Synthesis Research Articles

Оптимизация получения полисахаридов из Cyanobacterium sp. IPPAS B-1200

Cyanobacterium sp. IPPAS B-1200 is a cyanobacteria strain that belongs to the rare genus Cyanobacterium, family Cyanobacteriaceae fam. nov. Studies devoted to the isolation of secondary metabolites from this strain concentrate mostly on the fatty acid composition while the issue of isolating and identifying exopolysaccharides remains understudied. However, polysaccharides from cyanobacteria are of scientific and economic interest in the framework of biotechnology, medicine, pharmacology, etc. The research objective was to study the effect of the physicochemical conditions of cultivation and the composition of the cultural medium on exopolysaccharide production.
 Cyanobacterium sp. B-1200 were grown under 7500 ± 50 lux (12 h light/12 h dark). The dry cell weight was determined by gravimetry and a calibration plot that illustrated the dependence of the biomass amount on the degree of absorption at a wavelength of 750 nm. The amount of polysaccharides in the culture liquid was assessed by the Anthrone-sulphate method. The extraction was carried out by alcohol precipitation. The method of ultrasonic dispersion was used to destroy the cell walls of cyanobacteria. 
 The experimental study revealed the optimal parameters for the extraction and purification of exopolysaccharides from the culture medium. Removal of sodium bicarbonate from the medium and a 300%-increase in its concentration raised the yield of polysaccharides. The optimal value of active acidity for the synthesis of polysaccharides was pH = 6 while the optimal temperature for their accumulation was 35°C. The largest amount of biomass was obtained at 25°C. Intense white illumination contributed to the greatest release of exopolysaccharides into the culture medium; red-white illumination affected the morphology of cyanobacteria cells. During the extraction, the concentration, temperature, and nature of the extractant proved to be the most important factors. For example, isopropanol produced the highest yield while butanol triggered the least effective response. 
 The optimal extraction and purification modes for polysaccharides were as follows. For ultrasonic processing, the best results were obtained at a power of 20 W after 5 min. For freeze drying, the rational parameters were 8 h at –15°C.

Influence of Organic and Inorganic Compounds of Various Metals on the Synthesis of Polysaccharides by the Medicinal Mushroom Trametes versicolor.

To date, basidiomycetes are considered to be promising objects of biotechnology, due to a number of biologically active compounds, such as polysaccharides and triterpenes. These compounds have a high therapeutic potential and demonstrate immunomodulatory, antiviral, and antifungal activities. The purpose of this study was to study the effect of various concentrations of metal citrates and sulphates on the content of exo- and endopolysaccharides of the fungus Trametes versicolor. The mycelium was grown by deep cultivation on a semisyntheticglucose-peptone-yeast medium with different contents of zinc, copper, and manganese salts, after which the extraction and measurement of the concentration of polysaccharides were carried out. The results obtained showed that copper citrate at a concentration of 4 mg/L had the greatest positive effect on biomass yield. The intensity of biomass growth on a nutrient medium with copper citrate increased by 80%. Zinc citrate increased the content of exopolysaccharides by 29% compared to the medium without metal salts. When manganese citrate was added to the medium, the productivity of synthesis decreased, but an increase in the growth rate of mycelium biomass was observed. Sulphates of these metals led to a decrease in the productivity of exopolysaccharide synthesis by 12% for zinc and 35% for manganese. The addition of both copper citrate and copper sulphate to the medium led to a decrease in the synthesis productivity by 66 and 24%, respectively. The introduction of both citrates and sulphates of these metals into the culture medium led to an increase in the percentage of endopolysaccharides in the mycelium of the fungus. Copper citrate enhances Trametes versicolor biomass by 80%. Zinc citrate increases exopolysaccharide content by 29%. Copper sulphate optimizes endopolysaccharide production.

Genomic characterization of rare earth binding by Shewanella oneidensis

Rare earth elements (REE) are essential ingredients of sustainable energy technologies, but separation of individual REE is one of the hardest problems in chemistry today. Biosorption, where molecules adsorb to the surface of biological materials, offers a sustainable alternative to environmentally harmful solvent extractions currently used for separation of rare earth elements (REE). The REE-biosorption capability of some microorganisms allows for REE separations that, under specialized conditions, are already competitive with solvent extractions, suggesting that genetic engineering could allow it to leapfrog existing technologies. To identify targets for genomic improvement we screened 3,373 mutants from the whole genome knockout collection of the known REE-biosorbing microorganism Shewanella oneidensis MR-1. We found 130 genes that increased biosorption of the middle REE europium, and 112 that reduced it. We verified biosorption changes from the screen for a mixed solution of three REE (La, Eu, Yb) using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) in solution conditions with a range of ionic strengths and REE concentrations. We identified 18 gene ontologies and 13 gene operons that make up key systems that affect biosorption. We found, among other things, that disruptions of a key regulatory component of the arc system (hptA), which regulates cellular response to anoxic environments and polysaccharide biosynthesis related genes (wbpQ, wbnJ, SO_3183) consistently increase biosorption across all our solution conditions. Our largest total biosorption change comes from our SO_4685, a capsular polysaccharide (CPS) synthesis gene, disruption of which results in an up to 79% increase in biosorption; and nusA, a transcriptional termination/anti-termination protein, disruption of which results in an up to 35% decrease in biosorption. Knockouts of glnA, pyrD, and SO_3183 produce small but significant increases (≈ 1%) in relative biosorption affinity for ytterbium over lanthanum in multiple solution conditions tested, while many other genes we explored have more complex binding affinity changes. Modeling suggests that while these changes to lanthanide biosorption selectivity are small, they could already reduce the length of repeated enrichment process by up to 27%. This broad exploratory study begins to elucidate how genetics affect REE-biosorption by S. oneidensis, suggests new areas of investigation for better mechanistic understanding of the membrane chemistry involved in REE binding, and offer potential targets for improving biosorption and separation of REE by genetic engineering.

Exopolysaccharide produced by Lactiplantibacillus plantarum Y12 exhibits inhibitory effect on the Shigella flexneri genes expression related to biofilm formation

Shigella is a specific enteric pathogen in humans, causing symptoms of bacterial dysentery. The biofilm formation of S. flexneri contributes to the emergence of multidrug resistance and facilitates the establishment of persistent chronic infections. This study investigated the regulatory effects of Lactiplantibacillus plantarum Y12 exopolysaccharide (L-EPS) on gene expression and its spatial hindrance effects in inhibiting the biofilm formation of S. flexneri. The transcriptome analysis revealed a significant impact of L-EPS on the gene expression profile of S. flexneri, with a total of 968 genes showing significant changes (507 up-regulated and 461 down-regulated). The significantly down-regulated KEGG metabolic pathway enriched in phosphotransferase system, Embden-Meyerhf-Parnas, Citrate cycle, Lipopolysaccharide biosynthesis, Cationic antimicrobial peptide resistance, Two-component system. Moreover, L-EPS significantly down-regulated the gene expression levels of fimbriae synthesis (fimF), lipopolysaccharide synthesis (lptE, lptB), anchor protein repeat domain (arpA), virulence factor (lpp, yqgB), antibiotic resistance (marR, cusB, mdtL, mdlB), heavy metal resistance (zraP), and polysaccharide synthesis (mtgA, mdoB, mdoC). The expression of biofilm regulator factor (bssS) and two-component system suppressor factor (mgrB) were significantly up-regulated. The RT-qPCR results indicated that a major component of L-EPS (L-EPS 2-1) exhibited the gene regulatory effect on the S. flexneri biofilm formation. Furthermore, electrophoresis and isothermal microtitration calorimetry demonstrated that the interaction between L-EPS 2-1 and eDNA is electrostatic dependent on the change in environmental pH, disrupting the stable spatial structure of S. flexneri biofilm. In conclusion, L-EPS inhibited the biofilm formation of S. flexneri through gene regulation and spatial obstruction effects.

Identification of potential biomarkers of myopia based on machine learning algorithms

PurposeThis study aims to identify potential myopia biomarkers using machine learning algorithms, enhancing myopia diagnosis and prognosis prediction.MethodsGSE112155 and GSE15163 datasets from the GEO database were analyzed. We used “limma” for differential expression analysis and “GO plot” and “clusterProfiler” for functional and pathway enrichment analyses. The LASSO and SVM-RFE algorithms were employed to screen myopia-related biomarkers, followed by ROC curve analysis for diagnostic performance evaluation. Single-gene GSEA enrichment analysis was executed using GSEA 4.1.0.ResultsThe functional analysis of differentially expressed genes indicated their role in carbohydrate generation and polysaccharide synthesis. We identified 23 differentially expressed genes associated with myopia, four of which were highly effective diagnostic biomarkers. Single gene GSEA results showed these genes control the ubiquitin-mediated protein hydrolysis pathway.ConclusionOur study identifies four key myopia biomarkers, providing a foundation for future clinical and experimental validation studies.

Genome Analysis and Potential Ecological Functions of Members of the Genus Ensifer from Subsurface Environments and Description of Ensifer oleiphilus sp. nov.

The current work deals with genomic analysis, possible ecological functions, and biotechnological potential of two bacterial strains, HO-A22T and SHC 2-14, isolated from unique subsurface environments, the Cheremukhovskoe oil field (Tatarstan, Russia) and nitrate- and radionuclide-contaminated groundwater (Tomsk region, Russia), respectively. New isolates were characterized using polyphasic taxonomy approaches and genomic analysis. The genomes of the strains HO-A22T and SHC 2-14 contain the genes involved in nitrate reduction, hydrocarbon degradation, extracellular polysaccharide synthesis, and heavy metal detoxification, confirming the potential for their application in various environmental biotechnologies. Genomic data were confirmed by cultivation studies. Both strains were found to be neutrophilic, chemoorganotrophic, facultatively anaerobic bacteria, growing at 15-33 °C and 0-1.6% NaCl (w/v). The 16S rRNA gene sequences of the strains were similar to those of the type strains of the genus Ensifer (99.0-100.0%). Nevertheless, genomic characteristics of strain HO-A22T were below the thresholds for species delineation: the calculated average nucleotide identity (ANI) values were 83.7-92.4% (<95%), and digital DNA-DNA hybridization (dDDH) values were within the range of 25.4-45.9% (<70%), which supported our conclusion that HO-A22T (=VKM B-3646T = KCTC 92427T) represented a novel species of the genus Ensifer, with the proposed name Ensifer oleiphilus sp. nov. Strain SHC 2-14 was assigned to the species 'Ensifer canadensis', which has not been validly published. This study expanded the knowledge about the phenotypic diversity among members of the genus Ensifer and its potential for the biotechnologies of oil recovery and radionuclide pollution treatment.

Biological Properties and Antimicrobial Potential of Cocoa and Its Effects on Systemic and Oral Health.

Cocoa is considered a functional food because it is a natural source of macro- and micronutrients. Thus, cocoa is rich in vitamins, minerals, fiber, fatty acids, methylxanthines and flavonoids. In addition to favoring the metabolism of lipids and carbohydrates, the bioactive components of cocoa can have an antioxidant, anti-inflammatory and antimicrobial effect, providing numerous benefits for health. This literature review presents an overview of the effects of cocoa, fruit of the Theobroma cacao tree, on systemic and oral health. Several studies report that cocoa intake may contribute to the prevention of cardiovascular, neurodegenerative, immunological, inflammatory, metabolic and bone diseases, in addition to reducing the risk of vascular alterations and cognitive dysfunctions. On oral health, in vitro studies have shown that cocoa extract exerted an inhibitory effect on the growth, adherence and metabolism of cariogenic and periodontopathogenic bacteria, also inhibiting acid production, glycosyltransferase enzyme activity and the synthesis of insoluble polysaccharides. Additionally, administration of cocoa extract reduced biofilm accumulation and caries development in animals infected with cariogenic species. Clinical studies also reported that the use of mouthwashes containing cocoa extract reduced Streptococcus mutans counts in saliva and dental biofilm formation. In short, these studies highlight the nutritional value of cocoa, considering its clinical applicability, stability and economic accessibility.

Lysine lactylation regulates metabolic pathways and biofilm formation in Streptococcus mutans.

In eukaryotes, lactate produced during glycolysis is involved in regulating multiple metabolic processes through lysine lactylation (Kla). To explore the potential link between metabolism and Kla in prokaryotes, we investigated the distribution of Kla in the cariogenic bacterium Streptococcus mutans during planktonic growth in low-sugar conditions and in biofilm-promoting, high-sugar conditions. We identified 1869 Kla sites in 469 proteins under these two conditions, with the biofilm growth state showing a greater number of lactylated sites and proteins. Although high sugar increased Kla globally, it reduced lactylation of RNA polymerase subunit α (RpoA) at Lys173. Lactylation at this residue inhibited the synthesis of extracellular polysaccharides, a major constituent of the cariogenic biofilm. The Gcn5-related N-acetyltransferase (GNAT) superfamily enzyme GNAT13 exhibited lysine lactyltransferase activity in cells and lactylated Lys173 in RpoA in vitro. Either GNAT13 overexpression or lactylation of Lys173 in RpoA inhibited biofilm formation. These results provide an overview of the distribution and potential functions of Kla and improve our understanding of the role of lactate in the metabolic regulation of prokaryotes.

Necessity of Different Lignocellulose on Exopolysaccharide Synthesis and Its Hypoglycemic Activity In Vitro of Inonotus obliquus.

Current work proposes elm sawdust, poplar sawdust, pine sawdust, and cotton straw with different lignocellulose compositions and structures as the research objects to investigate the relationship between the hypoglycemic activity of mycelium polysaccharides from Inonotus obliquus and lignocellulose biodegradation. Four kinds of lignocellulosic materials could significantly increase the exopolysaccharide content and α-glucosidase inhibition rate and advance the occurrence time of α-glucosidase inhibition activity. Among all groups, the polysaccharide synthesis promotion effect of the cotton straw group was the best, which exopolysaccharide yield was 92.05% higher than that of the control group after 11-day fermentation. Meanwhile, the highest α-glucosidase inhibitory activity was found in the elm sawdust group on the 11th day (30.99%, which was 137.47% higher than control), and the exopolysaccharide in the elm sawdust group showed its effectiveness on glucose consumption of insulin resistant HepG2 cells at the concentration of 20 µg/mL, significantly higher than that of the metformin group (P < 0.05). The cellulose in the non-crystalline region of elm and pine and the hemicellulose of poplar were mainly used in the fermentation of I. obliquus, while the cellulose in the crystalline zone and amorphous zone of cotton straw was degraded to improve the exopolysaccharide content of I. obliquus. This paper revealed the necessity of different kinds of lignocellulose for the synthesis of active polysaccharide from I. obliquus and provided a new idea for the regulation of polysaccharide synthesis pathway.

General nitrogen regulation protein NtrC regulates the expression of nitrile hydratase and the synthesis of extracellular polysaccharide in Ensifer adhaerens CGMCC 6315

Ensifer adhaerens CGMCC 6315 contains two nitrile hydratases, CnhA and PnhA, which both convert the neonicotinoid insecticide acetamiprid to its amide metabolite. Here, we report that upregulation of PnhA and CnhA expression is related to nitrogen metabolism. Proteomic analysis showed that general nitrogen regulation protein NtrC was upregulated under nutrient deprivation. We thus constructed a ΔntrC mutant strain of E. adhaerens CGMCC 6315. In the ΔntrC strain, when cells were cultured in minimal salts medium supplemented with ammonium chloride, acetamiprid degradation and the expression of cnhA and pnhA were enhanced relative to those in wild-type E. adhaerens CGMCC 6315. In addition, NtrC negatively regulates synthesis of extracellular polysaccharides in this bacterium. The germination rate of soybean seeds was improved by inoculation with the ΔntrC strain of E. adhaerens CGMCC 6315 compared with the wild-type. This study is the first to report regulation of the expression of nitrile hydratase by NtrC, which is important for the practical remediation of pesticide pollution.

Transcriptomic analysis of biofilm formation by Bacillus cereus under different carbon source conditions

Abstract Background Previous studies found differences in the utilization of different carbon sources during biofilm formation by Bacillus cereus. Illumina HiSeq high-throughput sequencing technology was used to investigate the changes in gene transcript levels in Bacillus cereus biofilm bacteria under different carbon source conditions. Results Compared with the control group, the number of differentially expressed genes in the glucose, maltose, lactose, and skim milk-supplemented groups was 351, 1136, 133, and 487, respectively. The results showed that the pathways involved in the differentially expressed genes were mainly distributed in glycolysis and pentose phosphate pathway, tricarboxylic acid cycle, amino acid metabolism, and fatty acid metabolism. The gene expression of enzymes related to acetoin synthesis from pyruvate was mostly upregulated in the glucose-supplemented group. The gene expression of enzymes related to pyruvate synthesis of branched-chain amino acids in the maltose-supplemented group was mostly upregulated. In the lactose-supplemented group, the gene expression of acetoin biosynthesis from pyruvate was upregulated. Pyruvate production through glycolysis pathway increased in the skim milk-supplemented group, but the metabolic capacity of the tricarboxylic acid cycle did not change significantly. Conclusion The content of pyruvate stored by Bacillus cereus biofilm bacteria through glycolysis or pentose phosphate pathway increased, but the carbon flux into the tricarboxylic acid cycle did not increase, which suggested that carbon fluxes in the extracellular polysaccharide synthesis pathway of the biofilm may be increased, resulting in increased biofilm biomass formation.

Molecular Epidemiology genes detection of Klebsiella pneumoniae Clinical Isolates from the Adult Patients with Comorbidities in Baghdad hospitals

Background Klebsiella species is one of the most worldwide opportunistic pathogens in the world that cause infections at multiple sites like lung, urinary tract, bloodstream, wound or surgical site, and brain. Klebsiella pneumoniae has many types of virulence factors that give the bacteria the ability to invade the host and cause infections. Methods A total of (105) human clinical samples were collected from different patients with different cases. Klebsiella pneumonia were isolated from 30 samples. The bacteria were identified by biochemical tests and certified by using VITEK 2 system and genetically by amplify using PCR for 16 rRNA gene. One of the Klebsiella pneumoniae virulence factors is the capsule that is responsible for the macrophage resistance and its also responsible for serotype. So to confirm the strains isolated, PCR Gene detection of, MagA, K2A, RmpA, and Kfu, where (magA, k2A, rmpA, kfu) gens are responsible for capsule synthesis of Klebsiella pneumonia have been done. Results There were 30 strains of Klebsiella pneumoniae were identified. Epidemiological studies show that infections are preceded by gastrointestinal colonization, with the gastrointestinal tract being the most important reservoir for transmission. The primary bacterial isolates were identified based on their cultural, microscopic, and biochemical traits. The API system (API 20E) and VITEK 2 systems were used to confirm the identity of the bacterial isolates. Colony morphology revealed that Klebsiella pneumonia was Gram-negative, non-motile, microscopic straight rods grouped singly or in pairs. Biochemical tests showed that neutral end products prevailed over acidic end products, with Klebsiella producing negative results for indole. The Kligler Iron Agar (KIA) test showed that Klebsiella isolates changed the color of the slant and butt, producing an acidic slant and acid butt, along with gas production. The urease test for Klebsiella distinguishes between Enterobacter and Klebsiella isolates, as Klebsiella can manufacture the urease enzyme. The indole test distinguishes K. pneumonia from K. pneumonia from K. mobiliz. The API 20E system was used for biochemical testing, and the VITEK 2 system was used for identification. The VITEK 2 system was used to test for biochemical and antibiotic susceptibility, and the results supported the findings from morphological and biochemical analyses. PCR amplification was used for molecular detection, and all nine isolates tested positive for the magA and k2A genes. The ferric iron uptake system gene (kfu) and the extracapsular polysaccharide synthesis regulator gene (rmpA) were amplified using specific primer pairs. The VITEK 2 system demonstrated promising outcomes for K. pneumoniae identification. However, standard diagnostic methods are costly and time-consuming. Conclusions Navigating the complex landscape of infectious diseases, collaborative research projects have become indispensable in unraveling the mysteries surrounding pathogens. The project in Baghdad, having stood as a shining example of such collaboration, provided valuable insights into the identification, characterization, and molecular aspects of Klebsiella pneumoniae strains. The findings, have not only contributed to the scientific community's understanding of bacterial infections but also paved the way for enhanced strategies in diagnosis and treatment.

Antibacterial mechanism of areca nut essential oils against Streptococcus mutans by targeting the biofilm and the cell membrane.

Dental caries is one of the most common and costly biofilm-dependent oral diseases in the world. Streptococcus mutans is the major cariogenic pathogen of dental caries. S. mutans synthesizes extracellular polysaccharides by autologous glucosyltransferases, which then promotes bacterial adhesion and cariogenic biofilm formation. The S. mutans biofilm is the principal target for caries treatment. This study was designed to explore the antibacterial activity and mechanisms of areca nut essential oil (ANEO) against S. mutans. The ANEOs were separated by negative pressure hydro-distillation. The Kirby-Bauer method and broth microdilution method were carried out to evaluate the antibacterial activity of different ANEOs. The antibacterial mechanism was revealed by crystal violet staining, XTT reduction, microbial adhesion to hydrocarbon test, extracellular polysaccharide production assay, glucosyltransferase activity assay, lactate dehydrogenase leaking, propidium iodide staining and scanning electron microscopy (SEM). The cytotoxicity of ANEOs was determine by MTT assay. The ANEOs separated at different temperatures exhibited different levels of antibacterial activity against S. mutans, and the ANEO separated at 70°C showed the most prominent bacteriostatic activity. Anti-biofilm experiments showed that the ANEOs attenuated the adhesion ability of S. mutans by decreasing the surface hydrophobicity of the bacteria, prevented S. mutans biofilm formation by inhibiting glucosyltransferase activity, reducing extracellular polysaccharide synthesis, and reducing the total biofilm biomass and activity. SEM further demonstrated the destructive effects of the ANEOs on the S. mutans biofilm. Cell membrane-related experiments indicated that the ANEOs destroyed the integrity of the cell membrane, resulting in the leakage of lactic dehydrogenase and nucleic acids. SEM imaging of S. mutans cell showed the disruption of the cellular morphology by the ANEOs. The cytotoxicity assay suggested that ANEO was non-toxic towards normal oral epithelial cells. This study displayed that ANEOs exerted antibacterial activity against S. mutans primarily by affecting the biofilm and disrupting the integrity of the cell membrane. ANEOs has the potential to be developed as an antibacterial agent for preventing dental caries. Additionally, a new method for the separation of essential oil components is presented.

Pseudomonas composti isolate from oyster digestive tissue specifically binds with norovirus GII.6 via Psl extracellular polysaccharide

Oysters are recognized as important vectors for human norovirus transmission in the environment. Whether norovirus binds to bacteria in oyster digestive tissues (ODTs) remains unknown. To shed light on this concern, ODT-54 and ODT-32, positive for histo-blood group antigen (HBGA) -like substances, were isolated from ODTs and identified as Pseudomonas composti and Enterobacter cloacae, respectively. The binding of noroviruses (GII.4 and GII.6 P domains) to bacterial cells (ODT-32 and ODT-54; in situ assay) as well as extracted extracellular polysaccharides (EPSs; in vitro assay) was analyzed by flow cytometry, confocal laser scanning microscopy, ELISA, and gene knock-out mutants. ODT-32 bound to neither GII.4 nor GII.6 P domains, while ODT-54 specifically binds with GII.6 P domain through Psl, an exopolysaccharide encoded by the polysaccharide synthesis locus (psl), identified based on gene annotation, gene transcription, Psl specific staining, and ELISAs. These findings attest that ODT bacteria specifically bind with certain norovirus genotypes in a strain-dependent manner, contributing to a better understanding of the transmission and enrichment of noroviruses in the environment.

Mutations in type II Golgi-localized proton pyrophosphatase AVP2;1/VHP2;1 affect pectic polysaccharide rhamnogalacturonan-II and alter root growth under low boron condition in Arabidopsis thaliana.

The essential plant nutrient boron is required for the crosslinking of the pectin polysaccharide, rhamnogalacturonan II (RG-II). The synthesis of the pectic polysaccharides takes place in the Golgi apparatus, acidified by proton pumps. AVP2;1/VHP2;1 is a type II proton pyrophosphatase localized in the Golgi apparatus, which possesses proton pumping activity coupled with pyrophosphate hydrolysis. Its activity and expression patterns have been previously revealed but its role in plants remains unknown. The aim of the present work therefore was to explore the physiological role of AVP2;1 in Arabidopsis thaliana. In the screening of mutants under low boron, a mutant carrying a missense mutation in AVP2;1 was isolated. This mutant showed increased primary root growth under low boron conditions but no significant difference under normal boron condition compared to wild type plants. T-DNA insertion caused similar growth, suggesting that reduced function of AVP2;1 was responsible. Root cell observation revealed an increase in meristematic zone length, cell number in meristem and length of matured cell in avp2;1 mutants compared to wild type under low boron. Calcium concentration was reduced in mutant root cell wall under low boron. RG-II specific sugars also tended to be decreased in mutant root cell wall under low and normal boron conditions. These results suggest that changes in cell wall component by mutations in AVP2;1 may possibly explain the increased root length of mutants under low boron. This supports the idea that AVP2;1 plays a role in pH homoeostasis in Golgi apparatus for pectin synthesis.

Development of silver nanoparticles based on the method using quince seed mucilage for ascorbic acid determination.

Nanoparticles are used in various fields such as chemistry, pharmacy, biotechnology, and food science since they provide higher sensitivity than traditional optical detection methods. Recently, synthesis of nanomaterials using green chemistry has become popular. Many phytochemical components are used in the synthesis of nanoparticles, including vitamins, proteins, polysaccharides, glycosides, essential oils and phenolic compounds. A novel green nanotechnology-based method using quince seed mucilage (QSM) was designed for the determination of ascorbic acid in pharmaceutical preparations. QSM, a natural polysaccharide, was used as a bioreducing and stabilizing reagent in the proposed silver nanoparticle (SNP)-based method. In the first stage of the developed method, silver(I) is reduced to silver(0) via QSM and spherical, homogeneous SNPs were prepared (QSM-SNPs). In the second stage of the developed method, SNPs nuclei were enlarged with the addition of ascorbic acid. The developed method was validated by performance parameters (linearity, recovery, and precision). Ascorbic acid determination was performed by measuring increase in absorbance at 420 nm. The limit of detection and limit of quantification for ascorbic acid were, respectively, found to be at 0.27 and 0.90 μM. The QSM-SNP-based method was successfully applied to effervescent tablets containing ascorbic acid. The standards of the excipients frequently used in pharmaceutical preparations did not interfere with the developed method. The developed QSM-SNP-based method satisfies the requirements of green nanotechnology. The developed QSM-SNP-based method is simple, fast, eco-friendly and low-cost.

The mechanism of polysaccharide synthesis of Sanghuangporus sanghuang based on multi-omic analyses and feedback inhibition

S. sanghuang polysaccharide has various biological roles in promoting human health, however, the underlying mechanism of polysaccharide synthesis in S. sanghuang remain elusive. In the present study, the molecular structure of novel polysaccharide in the mutant S. sanghuang strain A130 with high yield of polysaccharide was characterized. The critical genes/proteins and pathways involved in polysaccharide synthesis were investigated via comparative transcriptomic, proteomic, and integrative analysis between wildtype strain SH-1 and A130. An integrated analysis of transcriptomic and proteomic results was also performed to locate potential regulators in the production of polysaccharides. The genes of cellobiohydrolase1 (CBH1) and MutS Homolog 6 (MSH6) related to glycolysis/gluconeogenesis were differentially expressed between A130 and SH-1, suggesting the potential involvement of these genes in regulating the production of polysaccharide. Proteomic analysis revealed that the abundance of Tyrosinase (TYR) and Trehalase (TREH) were substantially different between A130 and SH-1. The potential involvement of TYR in polysaccharide production was confirmed by transcriptomic-proteomic integrated analysis. The biological role of TYR and TREH in polysaccharide production was further verified by feedback inhibition of kojic acid and validamycin A, respectively. Overall, our study provides critical insights for the polysaccharide synthesis and high yield of polysaccharide through genes/pathways regulating in S. sanghuang.

Development of lipid nanoparticles(LNP)- RNA vaccine against Streptococcus pneumonae gene cluster facilitating the synthesis of the capsular polysaccharide in Egypt.

Development of lipid nanoparticles(LNP)- RNA vaccine against Streptococcus pneumonae gene cluster facilitating the synthesis of the capsular polysaccharide in Egypt.

Alterations in Carbohydrate Metabolism and Modulation of Thermo-Tolerance in Tomato under Heat Stress

Heat stress affects the source and sink activities, growth and development, economic yield and harvest index of plants negatively. Heat stress has got severe effects on the biochemical reactions of photosynthesis which included irreversible damages of RuBisCO, oxygen evolving complexes, PSII reaction centres and disruptions of chloroplast ultrastructure and thylakoid membrane. Sugars are products of photosynthesis in plants which serve as substrates for energy metabolism and are required for synthesis of cellulose and starch polysaccharides. Glucose, fructose, sucrose, mannose, trehalose, maltose etc. are the sugars present in plants. Abiotic stresses like temperature, drought, flood and salinity causes crucial differences in the carbohydrate metabolism. Sugars acts as primary signaling molecules and regulate signals that control the expression of many genes and enzymes involved in carbohydrate metabolism. Decrease in crop productivity under heat stress is chiefly associated with alterations in carbohydrate accumulation and resulting defects in assimilation partitioning from source to sink. It is essential to understand the responses and acclimatization processes to extreme temperatures in plants, such that the mechanisms underlying can be used for developing heat-tolerant varieties. Sugars such as glucose, sucrose, fructan, raffinose and trehalose not only acts structural component and metabolic resources also regulate genes associated with heat stress tolerance and reduce the chances of crop loss. The external application of compounds like osmoregulators, phytohormones, signaling molecules, etc., have exhibited positive responses in stress tolerance due to its growth promoting and antioxidant activities.

Modified Lactoperoxidase System as a Promising Anticaries Agent: In Vitro Studies on Streptococcus mutans Biofilms.

The lactoperoxidase (LPO) system shows promise in the prevention of dental caries, a common chronic disease. This system has antimicrobial properties and is part of the non-specific antimicrobial immune system. Understanding the efficacy of the LPO system in the fight against biofilms could provide information on alternative strategies for the prevention and treatment of caries. In this study, the enzymatic system was modified using four different (pseudo)halide substrates (thiocyanate, thiocyanate-iodide mixture, selenocyanate, and iodide). The study evaluated the metabolic effects of applying such modifications to Streptococcus mutans; in particular: (1) biofilm formation, (2) synthesis of insoluble polysaccharides, (3) lactate synthesis, (4) glucose and sucrose consumption, (5) intracellular NAD+ and NADH concentrations, and (6) transmembrane glucose transport efficiency (PTS activity). The results showed that the LPO-iodide system had the strongest inhibitory effect on biofilm growth and lactate synthesis (complete inhibition). This was associated with an increase in the NAD+/NADH ratio and an inhibition of glucose PTS activity. The LPO-selenocyanate system showed a moderate inhibitory effect on biofilm biomass growth and lactate synthesis. The other systems showed relatively small inhibition of lactate synthesis and glucose PTS but no effect on the growth of biofilm biomass. This study provides a basis for further research on the use of alternative substrates with the LPO system, particularly the LPO-iodide system, in the prevention and control of biofilm-related diseases.