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This study identified a rhamnose-containing cell wall polysaccharide (RhaCWP) in an alkaline extract prepared to analyze intracellular polysaccharides (IPS) from Streptococcus mutans biofilm. IPS was an 1,4-α-D-glucan with branchpoints introduced by 1,6-α-glucan while RhaCWP presented 1,2-α-L-and 1,3-α-L rhamnose backbone and side chains connected by 1,2-α-D-glucans, as identified by nuclear magnetic resonance (NMR) spectroscopy and methylation analyses. The MW of IPS and RhaCWP was 11,298 Da, as determined by diffusion-ordered NMR spectroscopy. Therefore, this study analyzed the chemical structure of RhaCWP and IPS from biofilm in a single fraction prepared via a convenient hot-alkali extraction method. This method could be a feasible approach to obtain such molecules and improve the comprehension of the structure-function relationships in polymers from S. mutans in future studies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The Streptococcus genus comprises both commensal and pathogenic species. Additionally, Streptococcus thermophilus is exploited in fermented foods and in probiotic preparations. The ecological and metabolic diversity of members of this genus is matched by the complex range of cell wall polysaccharides that they present on their cell surfaces. These glycopolymers facilitate their interactions and environmental adaptation. Here, current knowledge on the genetic and compositional diversity of streptococcal cell wall polysaccharides including rhamnose-glucose polysaccharides, exopolysaccharides and teichoic acids is discussed. Furthermore, the species-specific cell wall polysaccharide combinations and specifically highlighting the presence of rhamnose-glucose polysaccharides in certain species, which are replaced by teichoic acids in other species. This review highlights model pathogenic and non-pathogenic species for which there is considerable information regarding cell wall polysaccharide composition, structure and genetic information. These serve as foundations to predict and focus research efforts in other streptococcal species for which such data currently does not exist., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Monosaccharides are essential for maintaining the normal physiological functions of living organisms. Under disease states, metabolic disorders in vivo will inevitably affect the levels of monosaccharides, which brings the possibility of monosaccharides as a biomarker of some diseases. In this study, a method was developed and validated for simultaneously determining 10 monosaccharides (glucose, galactose, mannose, rhamnose, fucose, xylose, iduronic acid, glucuronic acid, N-acetylgalactosamine and N-acetylglucosamine) in SD rat plasma using liquid chromatography-tandem mass spectrometry. The method employed 1-phenyl-3-methyl-5-pyrazolone (PMP) as a derivatization reagent, considerably improved the chromatographic retention and ionization efficiency of monosaccharides. After protein precipitation of plasma samples, monosaccharides and isotope internal standards were derivatized and liquid-liquid extraction was performed to remove excess PMP. To achieve the baseline separation of several isomers, the resulting derivatives were chromatographed on a Bridged ethyl hybrid (BEH) Phenyl column using gradient elution with a total run time of 8 min. The method was linear within the range of 0.0100-5.00 μg/mL for rhamnose, 0.0500-25.0 μg/mL for fucose, xylose, iduronic acid, glucuronic acid, N-acetylgalactosamine and N-acetylglucosamine, 1.00-500 μg/mL for galactose, 10.0-5000 μg/mL for mannose, and 50.0-25,000 μg/mL for glucose. And the accuracy and precision verification of surrogate matrix samples and plasma samples met the required criteria. The method has been used successfully to study the effect of hepatic insufficiency on monosaccharide levels in rats. It was found that the concentration of glucuronic acid in SD rat plasma was abnormally increased in rats with liver injury., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Xiaoyan Chen reports financial support was provided by The High-level new R&D institute (2019B090904008) and the High-level Innovative Research Institute (2021B0909050003) of the Department of Science and Technology of Guangdong Province., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The extraction, characterization and antioxidant activity of polysaccharides from Choerospondias axillaris leaves were investigated in the present study. Two purified polysaccharide fractions, CALP-1 and CALP-2, were isolated from crude Choerospondias axillaris leaf polysaccharides (CALP) by DEAE-52 cellulose chromatography and Sephadex G-100 column chromatography. The characteristics of CAL-1 and CALP-2 were determined by using High-performance Gel Permeation Chromatography (HPGPC), High-Performance Anion-Exchange Chromatography, HPAEC (HPAEC-PAD) and Fourier transform infrared spectroscopy (FTIR). CALP-1 with molecular weight of 11.20 KDa was comprised of Rhamnose, Arabinose, Galactose, Glucose, Xylose, Mannose and galacturonic acid in a molar ratio of 5.16:2.31:5.50:27.18:1.00:0.76:1.07. CAL-2 with molecular weight of 8.03 KDa consisted of Rhamnose, Arabinose, Galactose, Glucose, and galacturonic acid at a ratio of 1.38:3.63:18.84:8.28:1.45. FTIR revealed that CALP-1 and CALP-2 were acidic polysaccharides. The antioxidant activity of crude CALP, CALP-1 and CALP-2 was evaluated in vitro. The fraction CALP-2 was demonstrated to be of polysaccharide nature containing a large percentage of Galactose but no Xylose and Mannose. The antioxidant activity assays showed that CALP-1 and CALP-2 exhibited antioxidant and scavenging activities on hydroxyl and DPPH radicals in vitro. Compared with pure polysaccharide, crude CALP exhibited stronger anti-oxidant activities. These results will provide a better understanding of Choerospondias axillaris leaf polysaccharide and promote the potential applications of Choerospondias axillaris leaf polysaccharide in the pharmacological field and as a natural antioxidant.

Although the fruit of Ficus tikoua Bur. has been consumed by montanic people in China for centuries, its chemical and biological composition was still unclear. A series of comprehensive investigations on its chemical constituents and bioactivities were carried out for the first time. As a result, six compounds were isolated and identified as the main components in this fruit. GC-MS analysis of the lipid components demonstrated that Ficus tikoua Bur. fruit contains some wholesome constituents such as fatty acids, vitamins, triterpenoids, and phytosterols. The fatty acids are mainly composed of linolenic acid (61.27%) and linoleic acid (22.79%). Furthermore, this fruit contains a relative high content of crude protein (9.41 ± 0.03%), total amino acids (9.28%), and total polyphenols (0.86 ± 0.01 g/100 g). The analysis of monosaccharide composition showed that the total polysaccharide mainly consists of glucose, glucuronic acid, xylose, arabinose, mannose, galactose, galacturonic acid, and rhamnose. The polysaccharide, polyphenol, water, ethanol, and flavonoid extracts exhibited prominent antioxidant activity determined by ABTS, DPPH, and FRAPS methods. Meanwhile, the total polysaccharide exhibited significant immunomodulatory effect by enhancing the release of cytokines and expression of iNOS and COX-2 in RAW264.7 cells, significantly decreasing the expression of c-Jun and p65 proteins in the cytoplasm; increasing the translocation of c-Jun and p65 to the nucleus; and regulating the phosphorylation level of Akt, PI3K, and PDK1 in the PI3K/AKT signaling pathway. This study proved that the fruit of F. tikoua is a reliable source of functional food.

In this work we have efficiently extracted and characterized pectin from different tissues of astringent (AS) and non-astringent (NAS) persimmon fruits (peel, pulp, whole fruit) for the first time. The highest pectin extraction (≥7.2%) was carried out at 80 °C, 120 min with 1.5% sodium citrate in peel of both AS and NAS persimmon samples. All persimmon pectins showed a molecular weight and galacturonic acid content upper than 328 kDa and 78%, respectively, indicating their suitability as food ingredient. Pectin extracted from AS pulp and peel tissues exhibited an enriched structure in rhamnose and arabinose, whereas the opposite behavior was observed in NAS persimmon whole fruit samples. Remarkably, both pulp tissues (AS and NAS) presented the highest levels of glucose and mannose, non-pectic carbohydrates. In addition, techno-functional assessment (zeta potential, particle size, apparent viscosity, gelation) showed the suitability of the persimmon pectins for a broad range of industrial applications., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The aim of the present work is to isolate a series of triterpene derivatives with rhamnosyl linking acetyl groups from Glechoma longituba according to the structural characteristics of previously described triterpene saponins. The extract ion chromatography spectrum of the crude extract of G. longituba was detected and analyzed by HPLC-HR-ESI-MS to determine possible components, and these metabolites were traced and separated by combining high-resolution mass spectrometry and predicted liquid chromatography retention time. Three 11α, 12α-epoxypentacyclic oleanolic acid triterpene saponins (glechomanosides H-J) and one ursane triterpene aldehyde saponin with a C-28 aldehyde group were isolated from G. longituba. The structure of these compounds was confirmed by NMR and compared with those of previously characterized compounds. The strategy described in this report enables a rapid, reliable, and complete analysis of glycoside compounds containing different numbers of acetyl groups at different positions on the sugar., (© 2021 Wiley-VHCA AG, Zurich, Switzerland.)

An immuno-stimulatory polysaccharide (EtISPFa) was purified from water extract of the fungus Echinodontium tinctorium. EtISPFa has an estimated weight average molecular weight (M w ) of 1354 kDa and is composed of glucose (66.2 %), glucuronic acid (10.1 %), mannose (6.7 %), galactose (6.4 %), xylose (5.6 %), rhamnose (3.1 %), fucose (1.8 %), and arabinose (0.2 %). It has multiple glycosidic linkages, with 3-Glcp (19.8 %), 4-GlcpA (10.8 %), 6-Glcp (10.7 %), and 3,6-Glcp (8.7 %) being the most prominent. NMR analysis showed that EtISPFa has a backbone containing mostly of 3-substituted β-glucopyranose with 4-substituted glucopyranosyluronic acid. Short side chains consisting of an average of two β-glycopyranose residues, connected through 1→6 linkages, are attached to the 6-position of about every 4th or 5th backbone glucose residue. EtISPFa is a novel glucuronic acid-containing β-glucan capable of significantly inducing the production of cytokines IL-17, IL-16, MIP-2, G-CSF,GM-CSF, LIF, MIP-1α, MIP-1β, and RANTES in vitro. EtISPFa should be further explored for its immuno-stimulatory activity in vivo., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Plant cells are encapsulated by cell walls whose properties largely determine cell growth. We have previously identified the rol1-2 mutant, which shows defects in seedling root and shoot development. rol1-2 is affected in the Rhamnose synthase 1 ( RHM1 ) and shows alterations in the structures of Rhamnogalacturonan I (RG I) and RG II, two rhamnose-containing pectins. The data presented here shows that root tissue of the rol1-2 mutant fails to properly differentiate the cell wall in cell corners and accumulates excessive amounts of callose, both of which likely alter the physical properties of cells. A surr ( suppressor of the rol1-2 root developmental defect ) mutant was identified that alleviates the cell growth defects in rol1-2 . The cell wall differentiation defect is re-established in the rol1-2 surr mutant and callose accumulation is reduced compared to rol1-2 . The surr mutation is an allele of the cyclin-dependent kinase 8 ( CDK8 ), which encodes a component of the mediator complex that influences processes central to plant growth and development. Together, the identification of the surr mutant suggests that changes in cell wall composition and turnover in the rol1-2 mutant have a significant impact on cell growth and reveals a function of CDK8 in cell wall architecture and composition.

Purpose: Develop a quantitative LC-MS/MS method for FDG, FDG-monophosphate, glucose and glucose-monophosphate in mouse tumor models to assist in validating the use of [ 18 F]FDG-positron emission tomography (PET) imaging for anticancer therapies in a clinical setting. Methodology/results: Analytes were isolated from tumors by protein precipitation and detected on a Sciex API-5500 mass spectrometer. Improved assay robustness and selectivity were achieved through chromatographic separation of FDG-monophosphate from glucose-monophosphate, selection of a unique ion transition and incorporation of stable isotope labeled internal standards. In a mouse JIMT-1 tumor model, FDG-monophosphate levels measured by LC-MS/MS correlated with [ 18 F]FDG-PET imaging results. Conclusion: LC-MS/MS analysis of FDG-monophosphate accumulation in tumors is a cost-effective tool to gauge the translational potential of [ 18 F]FDG-PET imaging as a noninvasive biomarker in clinical studies.

Simple, rapid, and accurate detection methods for saccharides are potentially applicable to various fields such as clinical and food chemistry. However, the practical applications of on-site analytical methods are still limited. To this end, herein, we propose a 96-well microtiter plate made of paper as a paper-based chemosensor array device (PCSAD) for the simultaneous classification of 12 saccharides and the quantification of fructose and glucose among 12 saccharides. The mechanism of the saccharide detection relied on an indicator displacement assay (IDA) on the PCSAD using four types of catechol dyes, 3-nitrophenylboronic acid, and the saccharides. The design of the PCSAD and the experimental conditions for the IDA were optimized using a central composite design. The chemosensors exhibited clear color changes upon the addition of saccharides on the paper because of the competitive boronate esterification. The color changes were employed for the subsequent qualitative, semiquantitative, and quantitative analyses using an automated algorithm combined with pattern recognition for digital images. A qualitative linear discrimination analysis offered discrimination of 12 saccharides with a 100% classification rate. The semiquantitative analysis of fructose in the presence of glucose was carried out from the viewpoint of food analysis utilizing a support vector machine, resulting in clear discrimination of the various concentrations of fructose. Most importantly, the quantitative detection of fructose in two types of commercial soft drinks was also successfully carried out without sample pretreatments. Thus, the proposed PCSAD can be a powerful method for on-site food analyses that can meet the increasing demand from consumers for sensors of saccharides.

A polysaccharide (ESPS) purified from Eupolyphaga sinensis Walker by ion exchange chromatography and gel chromatography was investigated, including its structure characterization and antitumor activity. The results showed that ESPS was composed of rhamnose, fucose, arabinose, xylose, glucose, and galactose in a molar ratio of 7.4: 3.1: 13.9: 9.3: 39.7: 26.5, with the mean weight (Mw) of 2.14 × 10 4 Da; the main chain of ESPS was mainly composed of → 4) - α - D - Glcp - (1 → and → 3) - β - D - Galp - (1 →, and the side chains were connected to the main chain through the O-6 atom of glucose and O-4 and O-6 atom of galactose. In addition, ESPS promoted the lymphocyte proliferation and inhibited liver cancer cells growth through enhancing lymphocyte activity in vitro, mainly NK cells. Moreover, ESPS markedly stimulated immunity in H22-bearing mice by increasing the spleen and thymus indices and effectively inhibited H22 cell growth in vivo. These data indicated that ESPS was a polysaccharide component possessing high anti-hepatocellular carcinoma activity, representing a potential immunotherapy candidate for the treatment of liver cancer., Competing Interests: Declaration of competing interest The authors declare that no conflict of interest exists in the present study., (Copyright © 2020. Published by Elsevier B.V.)

In this study, a novel water-soluble polysaccharide (PVLP-1) was extracted and purified from Sacha inchi (Plukenetia volubilis L.) seeds and the structure, antioxidant and immunomodulatory activity of PVLP-1 were investigated. PVLP-1 (144 kDa) consisted of glucose (69.76%), mannose (14.86%), arabinose (10.53%), galactose (2.42%), ribose (1.23%), rhamnose (0.27%) and xylose (0.93%). PVLP-1 displayed characteristic polysaccharide bands in Fourier transform NMR spectra and infrared. The primary structure of PVLP-1 was a heteropolysaccharide with a backbone of (1 → 6)-linked glucose, sidechains of (1 → 4)-linked mannose, (1 → 4)-linked glucose and (1 → 3, 6)-linked mannose and a residue unit of →1)-linked arabinose as revealed the methylation analysis. PVLP-1 possessed good water-holding capacity (WHC), oil-holding capacity (OHC) and antioxidant capacities. Besides, PVLP-1 induced the proliferation of RAW264.7 cell and enhanced the expression of inflammatory cytokines IL-6, TNF-alpha(TNF-α) and IL-1 beta (IL-1β). The present study indicated that PVLP-1 possessed immune-enhancing bioactivities and could be functional food or adjuvant drug to improve biological immunity of immunodeficiency diseases and hypoimmunity., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

An efficient enzymatic hydrolysis method was developed and optimized for the degradation of auricularia auricula polysaccharide (AAP) and the degradation product of AAP was characterized. Cellulase was used for the degradation of AAP. The yield of reducing sugar and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging rate were used as indices to optimize the enzymatic hydrolysis of AAP, based on response surface methodology (RSM). The resulting optimal enzymolysis conditions were as follows: enzyme dosage, 13,500 U/g; enzymolysis temperature, 50 °C; and pH, 4.2. Under these conditions, the actual yield of reducing sugar was 16.50 mg/mL and the DPPH radical scavenging rate was 87.97%. The degradation product of AAP (C-EAAP) was homogeneous and contained alpha and beta glycoside bonds, but did not contain protein or nucleic acid. The molecular weight of the degradation product was 5.94 × 10 5  Da. Monosaccharide composition analysis revealed that C-EAAP was composed of mannose (57.1%), glucuronic acid (10.0%), rhamnose (0.4%), glucose (22.5%), galactose (2.9%), xylose (6.0%), and fucose (1.1%). The antioxidant activity of the polysaccharide indicated that C-EAAP had better antioxidant activity than AAP. The scavenging rates of C-EAAP for hydroxyl radicals (·OH) and superoxide anion radicals (O 2 - ·) were 1.65 and 1.90 times those of AAP., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The Opuntia ficus indica (L.) (OFI) is used as a nutritional and pharmaceutical agent in various dietary and value added products. This study underlines the possible use of native prickly pear cladode powder as a functional ingredient for health-promoting food production. To summarise, chemical characterization of polyphenols, minerals and soluble dietary fibre was performed; furthermore, the antioxidant activity and bioaccessibility of polyphenols and minerals were assessed. Eleven compounds between phenolic acids and flavonoids were identified, with piscidic acid and isorhamnetin derivatives being the most abundant. Opuntia's dietary fibre was mainly constituted of mucilage and pectin, and was composed of arabinose, galactose, glucose, mannose, rhamnose, and xylose sugars. The polyphenols' bioaccessibility was very high: piscidic acid at 200%, eucomic and ferulic acids >110% and flavonoids from 89% to 100%. The prickly pear cladode powder is also a source of minerals, as cations (calcium, sodium, potassium and magnesium) and anions (sulphate and chloride), with high magnesium bioaccessibilty (93%). OFI powder showed good capacity of radical scavenging measured by DPPH and ABTS methods, with 740 and 775 μmol Trolox/100 g OFI, respectively. Finally, the presented results allow the consideration of this natural product as a source of several essential nutrients, with a possible use in the food industry as a functional ingredient.

We obtained a new acidic soy hull polysaccharide (SHP-1) with a molecular weight (Mw) of 4.81 × 10 5  g/mol through ammonium oxalate and microwave assisted extraction. SHP-1 was mainly composed of galacturonic acid, galactose, rhamnose and arabinose (molar ratio = 46.59%:17.95%:14.77%:13.97%) with small amounts of fucose, glucose, mannose and xylose. The chemical structure was presumed to be of pectin-I type, consisting of 2/3 HGA and 1/3 RG-I. Furthermore, the rheological information and the chain morphology of SHP-1 were different in five solvents. Surfactant, salt and alkali solutions enhanced the solubility and flexibility of the polysaccharide, but the polysaccharide showed decreased fluidity under acidic conditions. The addition of ions and alkali increased the consistency coefficient of the solution, but the effect was far less than that of the cross-linking morphology. The structural and morphological information of purified SHP should aid in further study of its structure-function relationships and applications., Competing Interests: Declaration of competing interest None., (Copyright © 2020. Published by Elsevier B.V.)

The incidence of prostate cancer malignancy along with other cancer types is increasing worldwide, resulting in high mortality rate due to lack of effective medications. Moringa oleifera has been used for the treatment of communicable and non-communicable ailments across tropical countries, yet, little has been documented regarding its effect on prostate cancer. We evaluated the acute toxicity and apoptosis inducing effect of glucomoringin-isothiocyanate rich soluble extracts (GMG-ITC-RSE) from M. oleifera in vivo and in vitro, respectively. Glucomoringin was isolated, identified, and characterized using fundamental analytical chemistry tools where Sprague-Dawley (SD) rats, murine fibroblast (3T3), and human prostate adenocarcinoma cells (PC-3) were used for acute toxicity and bioassays experiments. GMG-ITC-RSE did not instigate adverse toxic reactions to the animals even at high doses (2000 mg/kg body weight) and affected none of the vital organs in the rats. The extract exhibited high levels of safety in 3T3 cells, where more than 90% of the cells appeared viable when treated with the extract in a time-dependent manner even at high dose (250 µg/mL). GMG-ITC-RSE significantly triggered morphological aberrations distinctive to apoptosis observed under microscope. These findings obviously revealed the putative safety of GMG-ITC-RSE in vivo and in vitro, in addition to its anti-proliferative effect on PC-3 cells.

Husk tomato (Physalis ixocarpa Brot. var. Rendidora) waste was evaluated as a source of specialized pectin, and pectin extracted from this waste was characterized physicochemically. Fruit was blanched for 10 or 15 min and extracted in 0.1 N HCl for 15 to 25 min. Extracted pectin was subjected to physicochemical analysis. For all extraction conditions, the percentage of anhydrogalacturonic acid exceeded 60%, indicating that husk tomato was a good source of pectin. The degree of esterification of pectin molecules was 63% to 91%. The amount of extracted pectin decreased with increasing extraction time. The apparent viscosity of husk tomato pectin showed the characteristic behavior of pseudoplastic fluids. Neutral sugars were identified, and the amounts of 6 sugars (fucose, rhamnose, arabinose, galactose, glucose, and xylose) were quantified. Sugars identified in husk tomato pectin and present in the Rhamnogalacturonan I region, arabinose, galactose, and rhamnose suggest a highly branched structure, which will influence its future applications. Molecular weight values were 542 to 699 kDa, exceeding molecular weight values reported for commercial citrus pectins from 134 to 480 kDa. The extraction process significantly (P < 0.05) influenced the physicochemical properties of pectin. Up to 19.8% from the total amount of pectin in the husk tomato was extracted by 10 min of blanching and 20 min of a more heat treatment. Our findings indicate that husk tomato can be a good alternative source of pectin having highly distinctive physicochemical characteristics., (© 2017 Institute of Food Technologists®.)

A major purpose of exploratory metabolic profiling is for the identification of molecular species that are statistically associated with specific biological or medical outcomes; unfortunately, the structure elucidation process of unknowns is often a major bottleneck in this process. We present here new holistic strategies that combine different statistical spectroscopic and analytical techniques to improve and simplify the process of metabolite identification. We exemplify these strategies using study data collected as part of a dietary intervention to improve health and which elicits a relatively subtle suite of changes from complex molecular profiles. We identify three new dietary biomarkers related to the consumption of peas (N-methyl nicotinic acid), apples (rhamnitol), and onions (N-acetyl-S-(1Z)-propenyl-cysteine-sulfoxide) that can be used to enhance dietary assessment and assess adherence to diet. As part of the strategy, we introduce a new probabilistic statistical spectroscopy tool, RED-STORM (Resolution EnhanceD SubseT Optimization by Reference Matching), that uses 2D J-resolved 1 H NMR spectra for enhanced information recovery using the Bayesian paradigm to extract a subset of spectra with similar spectral signatures to a reference. RED-STORM provided new information for subsequent experiments (e.g., 2D-NMR spectroscopy, solid-phase extraction, liquid chromatography prefaced mass spectrometry) used to ultimately identify an unknown compound. In summary, we illustrate the benefit of acquiring J-resolved experiments alongside conventional 1D 1 H NMR as part of routine metabolic profiling in large data sets and show that application of complementary statistical and analytical techniques for the identification of unknown metabolites can be used to save valuable time and resources.

The aim of this study was to investigate the effects of three Lactobacillus plantarum strains on in-vivo small intestinal barrier function and gut mucosal gene transcription in human subjects. The strains were selected for their differential effects on TLR signalling and tight junction protein rearrangement, which may lead to beneficial effects in a stressed human gut mucosa. Ten healthy volunteers participated in four different intervention periods: 7-day oral intake of either L. plantarum WCFS1, CIP104448, TIFN101 or placebo, proceeded by a 4 weeks wash-out period. Lactulose-rhamnose ratio (an indicator of small intestinal permeability) increased after intake of indomethacin, which was given as an artificial stressor of the gut mucosal barrier (mean ratio 0.06 ± 0.04 to 0.10 ± 0.06, p = 0.001), but was not significantly affected by the bacterial interventions. However, analysis in small intestinal biopsies, obtained by gastroduodenoscopy, demonstrated that particularly L. plantarum TIFN101 modulated gene transcription pathways related to cell-cell adhesion with high turnover of genes involved in tight- and adhesion junction protein synthesis and degradation (e.g. actinin alpha-4, metalloproteinase-2). These effects were less pronounced for L. plantarum WCFS1 and CIP104448. In conclusion, L. plantarum TIFN101 induced the most pronounced probiotic properties with specific gene transcriptional effects on repair processes in the compromised intestine of healthy subjects., Competing Interests: The authors declare no competing financial interests.

Lactobacillus casei is a Gram positive lactic acid bacterium used in dairy fermentations and present in the normal human gut microbiota. Certain strains are recognized as probiotics with beneficial effects on human and animal health. L. casei BL23 is a potential probiotic strain endowed with anti-inflammatory properties and a model strain widely used in genetic, physiological and biochemical studies. A number of bacterial cell surface polysaccharides have been shown to play a role in the immune modulation activities observed for probiotic lactic acid bacteria. In the present work, we purified the most abundant carbohydrate polymer of L. casei BL23 cell wall, a neutral wall polysaccharide (WPS) and established its chemical structure by periodate oxidation, methylation analysis and 2D NMR spectroscopy. The WPS of L. casei BL23 was shown to contain α-Rha, α-Glc, β-GlcNAc and β-GalNAc forming a branched heptasaccharide repeating unit (variant 1) with an additional partial substitution with α-Glc (variant 2). A modified non-reducing end octasaccharide, corresponding to a terminal unit of the WPS (variant 3), was also identified and allowed to define the biological repeating unit of the WPS. To our knowledge, this is the first report of the identification of a biological repeating unit based on a chemical evidence, in a cell wall polysaccharide of a Gram positive bacterial species., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

A structural characterization of polysaccharides from edible tropical fruit named starfruit (Averrhoa carambola L.) was carried out. After the purification steps, two homogeneous fractions were obtained. Fraction 50R was composed of rhamnose, arabinose, galactose and uronic acid in 4.3:56.2:37.4:2M ratio, respectively and fraction 10R was composed of rhamnose, arabinose, galactose and uronic acid in 2.8:65.8:28.5:3M ratio, respectively. Methylation and NMR spectroscopy analyses showed that these fractions are formed by pectic arabinogalactans, which contain (1→3), (1→6) and (1→3,6)-linked Galp units. The side chains have 3-O-, 5-O- and 3,5-di-O-linked α-Araf and nonreducing end-units of α-Araf, Arap, β-Galp and α-GlcpA. These arabinogalactans were linked to type I rhamnogalacturonans., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

A water-soluble polysaccharide, designated as ABPS-21, was isolated from the stem barks of Acanthopanax brachypus by hot-water extraction and purified by Cellulose DEAE-52 and Sephacryl S-300 HR gel-filtration chromatography, respectively. The homogeneity and molecular weight were determined using HPGPC. The structure was elucidated based on monosaccharide composition and methylation analysis, partial acid hydrolysis, periodate oxidation, Smith degradation, IR and NMR spectroscopy. The result showed that ABPS-21 was a homogeneous heteropolysaccharide including galactose, glucose, rhamnose and galacturonic acid in a molar ratio of 3.0:2.0:2.0:1.0, respectively, with an average molecular weight of 1.06 × 10(5)Da. The main chain of ABPS-21 was made up of → 4)-β-D-Galp-(1 → 4)-β-D-Glcp-(1 → 4)-β-D-Galp-(1 → 4)-β-D-Galp-(1 → 2)-α-L-Rhap-(1 →, and two side chains α-l-Rhap-(1 → 4)-β-D-Glcp-(1 → and α-D-galpA-(1 → were attached to the backbone chain at O-6 position of 1,4,6-linked β-D-Glcp and 1,4,6-linked β-D-Galp, respectively. ABPS-21, an acidic branched polysaccharide from A. brachypus, was not described up to now., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Pseudomonas stutzeri AS22, when grown on media containing starch and yeast extract and incubated at 30 °C and 200 rpm for 24h, was found to produce an acidic and high-molecular mass exopolysaccharide (EPS22). The EPS22 was purified and a yield of 1.3g/l was achieved. The average molecular mass of the EPS22 was determined by high-performance size-exclusion chromatography (HPSEC) and showed an average molecular mass of 9.9 × 10(5)Da and a polydispersity index Mw/Mn (Mw, weight-average and Mn, number-average) of 1.197 ± 0.015. Structural data of this EPS22 were determined using a combination approach including monosaccharide composition (HPAEC-PAD and GLC), methylation analysis (GC-MS) and NMR spectroscopy analysis. EPS22 was found to be a complex heteropolysaccharide with a repeating unit mainly composed of glucose, mannose and lactyl rhamnose in a molar ratio of 1:1.1:0.7. The acidic nature of the polysaccharide is due to the presence of three non-osidic substituents consisting of a lactyl, acetyl, and pyruvyl groups., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In this study, a new bioflocculant (ZZ-3) is isolated and evaluated. This novel flocculant was derived Klebsiella, which was identified by 16S rDNA sequencing as well as biochemical and physiological analyses. The composition of ZZ-3 was found to be 84.6% polysaccharides and 6.1% protein. More specifically, the amount (moles) of the polysaccharides rhamnose, mannose, and galactose were found to be 6.48, 2.47, and 1.74 greater than glucose, respectively. Results show ZZ-3 has a relatively high molecular weight (603-1820 kDa) and contains many functional groups (hydroxyl, amide, carboxyl, and methoxyl) that likely contribute to flocculation. The results of microscopic observation, zeta potential measurements, and ZZ-3 bioflocculant structure suggested that bridging was the main mechanism for flocculation with kaolin. Based on a high flocculation efficiency, thermal stability, pH tolerance and the ability to flocculate without additional cations, ZZ-3 shows potential for industrial application., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In this study, two polysaccharides (Elaeagnus angustifolia L. polysaccharide-1 (PEA-1) and PEA-2) were prepared from Elaeagnus angustifolia L. Then, the preliminary structure and antioxidant activities of all the samples were investigated. The results showed that the average molecular weights for PEA-1 and PEA-2 were 9113 and 5020 Da, respectively. And, PEA-1 was mainly composed of rhamnose, xylose, mannose, glucose, and galactose, respectively. The components of PEA-2 were rhamnose, mannose, glucose, and galactose, respectively. Moreover, the Antioxidant assays demonstrated that PEA-1 possessed of strong free radicals scavenging activity and hydroxyl radicals scavenging activities, suggesting that PEA-1 could potentially be used as natural antioxidant.

Background: Parts I and II of this series of papers identified several associations between the ease of milling and the chemical compositions of different chickpea seed fractions. Non-starch polysaccharides were implicated; hence, this study examines the free sugars and sugar residues., Results: Difficult milling is associated with: (1) lower glucose and xylose residues (less cellulose and xyloglucans) and more arabinose, rhamnose and uronic acid in the seed coat, suggesting a more flexible seed coat that resists cracking and decortication; (2) a higher content of soluble and insoluble non-starch polysaccharide fractions in the cotyledon periphery, supporting a pectic polysaccharide mechanism comprising arabinogalacturonan, homogalacturonan, rhamnogalalcturonan, and glucuronan backbone structures; (3) higher glucose and mannose residues in the cotyledon periphery, supporting a lectin-mediated mechanism of adhesion; and (4) higher arabinose and glucose residues in the cotyledon periphery, supporting a mechanism involving arabinogalactan-proteins., Conclusion: This series has shown that the chemical composition of chickpea does vary in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour. Seed coat strength and flexibility, pectic polysaccharide binding, lectins and arabinogalactan-proteins have been implicated. Increased understanding in these mechanisms will allow breeding programmes to optimise milling performance in new cultivars., (© 2013 Society of Chemical Industry.)

Background: A lot of interesting research has been undertaken to enhance the yield of exopolysaccharides (EPS) produced by lactic acid bacteria (LAB). The objective of this study was to determine the influence of casein hydrolysates (CH) with molecular weight less than 3 kDa on cell viability, EPS synthesis and the enzyme activity involved in EPS synthesis during the co-culturing of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus in MRS broth for 72 h at 37 ± 0.1 °C., Results: The highest EPS yield (150.1 mg L⁻¹) was obtained on CH prepared with papain (CHP) at 48 h. At 24 h, EPS were composed of galactose, glucose and rhamnose in a molar ratio of 1.0:2.4:1.5. The monosaccharide composition changed with extension of the fermentation time. The activities of α-phosphoglucomutase, uridine 5'-diphosphate (UDP)-glucose pyrophosphorylase and UDP-galactose 4-epimerase were associated with EPS synthesis. Moreover, the activities of β-phosphoglucomutase and deoxythymadine 5'-diphosphate (dTDP)-glucose pyrophosphorylase involved in rhamnose synthesis were very low at the exponential growth phase and could not be detected during other given periods., Conclusion: The influence of different CH (<3 kDa) on LAB viability, EPS production, EPS monomeric composition and activity levels of key metabolic enzymes was distinct. Besides, their influence was related to the distribution of amino acids., (© 2013 Society of Chemical Industry.)

Cyanobacteria are aquatic and photosynthetic microorganisms, which contribute up to 30% of the yearly oxygen production on the earth. They have the distinction of being the oldest known fossils, more than 3.5 billion years old, and are one of the largest and most important groups of bacteria on earth. Cyanobacteria are an emerging source of potentially pharmacologically active products and, among these, there are the lipopolysaccharides. Despite their significant and well documented activity, very little is known about the cyanobacteria lipopolysaccharides (LPS) structure. The aim of this work is to investigate the structure of the highly TLR4-antagonist lipopolysaccharide from the cyanobacterium Oscillatoria plankthotrix FP1. The LPS was purified and analysed by means of chemical analysis and 1H and 13C NMR spectroscopy. The LPS was then degraded by Smith degradation, HF and acetic acid hydrolyses. All the obtained products were investigated in detail by chemical analysis, NMR spectroscopy and by mass spectrometry. The LPS consists of a high molecular mass and very complex molecule lacking Kdo and heptose residues, where the polysaccharide chain is mainly constituted by a backbone of 3-substituted α-l-rhamnose units. The core region is rich in galacturonic acid and mannose residues. Moreover a glycolipid portion, similar to Gram-negative lipid A, was identified. This was built up of a non phosphorylated (1'→6) linked glucosamine disaccharide, acylated with 3-hydroxylated fatty acids. In particular 3-hydroxypentadecanoic and 3-hydroxyesadecanoic acids were found, together with esadecanoic and tetradecanoic ones. Finally the presence of a galacturonic acid residue at 6-position of the distal glucosamine in place of the Kdo residue is suggested., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Four water-soluble polysaccharides, FCp-1, FCp-2, FCp-3, and FCp-4 were obtained from finger citron fruits (Citrus medica L. var. sarcodactylis) by hot-water extraction and ethanol precipitation, followed by routine separation procedure. Based on the calibration curve, molecular weights of them were estimated to be 113.9, 32.6, 140.3, and 177.1 kDa respectively. The acid hydrolysis, methylation, IR, GC-MS, and NMR experiments were used for composition analysis. FCp-1 was a heteropolysaccharide composed of arabinose, galactose, glucose, rhamnose, and xylose, with a molar ratio of 3.0:7.0:4.1:1.0:1.5. FCp-2 and FCp-4 were →4)-α-D-GalpA(1→ linking galacturonan differ in molecular weights. FCp-3 was a →6)-α-D-Glcp(1→ linking glucan. According to the results of in vitro assays, FCp-3 showed significantly and moderately enhancing capacities toward the proliferation of splenocytes and thymocytes respectively. Thus, FCp-3 or analogs may have further use as immunomodulatory agents., (Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.)

Several reports have focused on utilization of post-harvest residues of crops, while neglecting those residues produced by mango processing. These residues represent a waste of nutrients and a source of environmental contaminants. Such by-products could be valuable sources of dietary fiber (DF), antioxidant compounds, and single carbohydrates. The aim of this study was to evaluate some functional properties (FP), and the content of DF and polyphenols (PP) of the peel and coarse material obtained from residues during the industrial processing of Ataulfo and Tommy Atkins mangoes. The total dietary fiber (TDF) content was about 225 mg/g and 387 mg/g (dry weight) for the coarse material and the peel, respectively, from which soluble dietary fiber represented 23 and 42%, respectively. The main neutral sugar identified was rhamnose, especially in peels; the klason lignin (KL) content was 92 mg/g, which highlights the Ataulfo peel (Ataulfo-P) and the Tommy Atkins peel (Tommy Atkins-P). The extractable PP content in Ataulfo-P was higher than in Tommy-Atkins-P, and interesting data for non-extractable PP were obtained in the residues. FP as swelling, water holding, oil holding, and glucose absorption in the residues was studied, obtaining better functional properties when compared to cellulose fiber. The results show that mango industrial by-products, mainly from the Ataulfo-P variety, could be used as ingredients in food products because of their functional properties as well as their DF and PP content.

The cell wall polysaccharides of Regina and Sunburst cherry varieties at two developmental stages were extracted sequentially, and their changes in monosaccharide composition and functional properties were studied. The loosely-attached pectins presented a lower d-galacturonic acid/rhamnose ratio than ionically-bound pectins, as well as lower thickening effects of their respective 2% aqueous solution: the lowest Newtonian viscosity and shear rate dependence during the pseudoplastic phase. The main constituents of the cell wall matrix were covalently bound pectins (probably through diferulate cross-linkings), with long arabinan side chains at the RG-I cores. This pectin domain was also anchored into the XG-cellulose elastic network. Ripening occurred with a decrease in the proportion of HGs, water extractable GGM and xylogalacturonan, and with a concomitant increase in neutral sugars. Ripening was also associated with higher viscosities and thickening effects, and to larger distribution of molecular weights. The highest firmness and compactness of Regina cherry may be associated with its higher proportion of calcium-bound HGs localized in the middle lamellae of cell walls, as well as to some higher molar proportion of NS (Rha and Ara) in covalently bound pectins. These pectins showed significantly better hydration properties than hemicellulose and cellulose network. Chemical composition and functional properties of cell wall polymers were dependent on cherry variety and ripening stage, and helped explain the contrasting firmness of Regina and Sunburst varieties., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Results of monosaccharide analysis of Ralstonia solanacearum lipopolysaccharides of the strains of different geographical origin are shown. Such monosaccharides as glucose, rhamnose and glucosamine prevail. Following the results of analysis, strains with supposedly truncated O-polysaccharide are revealed. As to their quantitative and monosaccharide composition of LPS the strains have demonstrated high level of heterogeneity.

A water-soluble arabinogalactan-protein (AGP), designated LRGP3, was isolated from the fruits of Lycium ruthenicum Murr. Its average molecular weight was 75.6 kDa. Monosaccharide composition analysis revealed that it was composed of rhamnose, arabinose, and galactose in a molar ratio of 1.0:14.9:10.4. Protein accounted for 1.7% of the AGP and was rich in hydroxyproline. On the basis of monosaccharide composition, partial acid hydrolysis, methylation analysis, ESI-MS, and NMR spectroscopy, LRGP3 was characterized as a highly branched polysaccharide with a backbone of (1→3)-linked β-D-galactopyranosyl residues, many of which were substituted at the O-6 position by galactosyl or arabinosyl groups. The branches were composed of (1→5)-linked arabinose, (1→2)-linked arabinose, (1→6)-linked galactose, (1→3)-linked galactose, and (1→2,4)-linked rhamnose, and the major nonreducing termini were α-L-arabinofuranosyl residues.

Calcein (R) could not only emit strong and stable room temperature phosphorescence (RTP) on filter paper using I(-) as perturber, but also could be oxidized by H(2)O(2) to form a non-phosphorescence compound (R'), resulting in the quenching of RTP signal of R. Moreover, the ortho-hydrogen of phenolic hydroxyl in R took condensation reaction with rhamnose (Rha) to produce non-phosphorescence compound (R-Rha) causing the RTP signal of R to further quench, and R-Rha was oxidized by H(2)O(2) to form R' and Rha, bringing about the sharp RTP signal quenching of R. Thus, a new solid substrate room temperature phosphorimetry (SSRTP) for the determination of trace Rha based on its strong catalytic effect on H(2)O(2) oxidizing R has been established, with the detection limit (LD) of 7.8zgspot(-1) (corresponding concentration: 2.0×10(-17) gm l(-1), sample volume: 0.40 μl spot(-1)). This method has been applied to determine trace Rha in cigarettes and jujubes, with the results coinciding well with those determined by a high performance liquid chromatography (HPLC). The component of R-Rha also was analyzed by means of HPLC, mass spectrometer and nuclear magnetic resonance (NMR) measurements. The mechanism of catalytic SSRTP for the determination of trace Rha was discussed., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Background: Cooking time decreases when beans are soaked first. However, the molecular basis of this decrease remains unclear. To determine the mechanisms involved, changes in both pectic polysaccharides and cell wall enzymes were monitored during soaking. Two cultivars and one breeding line were studied., Results: Soaking increased the activity of the cell wall enzymes rhamnogalacturonase, galactanase and polygalacturonase. Their activity in the cell wall was detected as changes in chemical composition of pectic polysaccharides. Rhamnose content decreased but galactose and uronic acid contents increased in the polysaccharides of soaked beans. A decrease in the average molecular weight of the pectin fraction was induced during soaking. The decrease in rhamnose and the polygalacturonase activity were associated (r = 0.933, P = 0.01, and r = 0.725, P = 0.01, respectively) with shorter cooking time after soaking., Conclusion: Pectic cell wall enzymes are responsible for the changes in rhamnogalacturonan I and polygalacturonan induced during soaking and constitute the biochemical factors that give bean cell walls new polysaccharide arrangements. Rhamnogalacturonan I is dispersed throughout the entire cell wall and interacts with cellulose and hemicellulose fibres, resulting in a higher rate of pectic polysaccharide thermosolubility and, therefore, a shorter cooking time., (Copyright © 2011 Society of Chemical Industry.)

A novel endophytic actinobacterium, designated strain EUM 378(T), was isolated from the surface-sterilized root tissue of Eucalyptus microcarpa, a eucalyptus tree known as Grey Box. Phylogenetic evaluation based on 16S rRNA gene sequence analysis, including alignment with taxon-specific 16S rRNA gene signature nucleotides, placed this isolate as a member of the family Nocardioidaceae. Strain EUM 378(T) showed >5.5 % 16S rRNA gene sequence divergence from other members of this family and was related most closely to Actinopolymorpha alba YIM 48868(T) (94.2 %) and Actinopolymorpha singaporensis IM 7744(T) (94.4 %). This Gram-positive, aerobic actinobacterium has well-developed substrate mycelia that fragment into small rods. Chemotaxonomic data revealed that the cell wall contains LL-diaminopimelic acid, ribose, glucose and rhamnose. MK-10(H₆) is the predominant menaquinone. Chemotaxonomic and phylogenetic evidence confirmed that strain EUM 378(T) represents a novel species of a new genus, for which the name Flindersiella endophytica gen. nov., sp. nov. is proposed. The type strain is EUM 378(T) ( = DSM 45355(T) = ACM 5289(T)).

The role of the BclA domains of B. cereus ATCC 14579 was investigated in order to understand the phenomena involved in the interfacial processes occurring between spores and inert surfaces. This was done by (i) creating deletions in the collagen-like region (CLR) and the C-terminal domain (CTD) of BclA, (ii) building BclA proteins with various lengths in the CLR and (iii) modifying the hydrophobic upper surface in the CTD. First, it was demonstrated that the CLR was substituted by three residues already reported in the CLR of B. anthracis, viz. rhamnose, 3-O-methyl-rhamnose, and GalNH(2) residues, while the CTD was also substituted by two additional glycosyl residues, viz. 2-O-methyl-rhamnose and 2,4-O-methyl-rhamnose. Regarding the properties of the spores, both CLR and CTD contributed to the adhesion of the spores, which was estimated by measuring the resistance to detachment of spores adhered to stainless steel plates). CLR and CTD also impacted the hydrophobic character and isoelectric point of the spores. It was then shown that the resistance to detachment of the spores was not affected by the physicochemical properties, but by the CLR length and the presence of hydrophobic amino acids on the CTD.

A Gram-stain-positive, non-spore-forming actinobacterium, designated 15-Je-017(T), was isolated from wall material of an indoor environment. The isolate formed a rudimentary substrate mycelium that fragmented into rod-shaped cells. On the basis of 16S rRNA gene sequence analysis, strain 15-Je-017(T) was shown to belong to the genus Jiangella and was most closely related to Jiangella alba YIM 61503(T) (99.7 % 16S rRNA gene sequence similarity), Jiangella alkaliphila D8-87(T) (99.0 %) and Jiangella gansuensis YIM 002(T) (99.0 %). The predominant menaquinone was MK-9(H(4)). Whole-cell hydrolysates contained ll-diaminopimelic acid as the diagnostic diamino acid in the cell wall and rhamnose and glucose as the main sugars. Mycolic acids were absent. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and seven unknown phospholipids. The fatty acid profile contained major amounts (>5 %) of anteiso-C(15 : 0), iso-C(16 : 0), iso-C(15 : 0), iso-C(17 : 0), anteiso-C(17 : 0) and C(17 : 1)ω8c, which supported the affiliation of strain 15-Je-017(T) to the genus Jiangella. DNA-DNA relatedness and physiological and biochemical tests allowed the differentiation of strain 15-Je-017(T) from the type strains of the three known Jiangella species. Strain 15-Je-017(T) represents a novel Jiangella species, for which we propose the name Jiangella muralis sp. nov., with type strain 15-Je-017(T) (=DSM 45357(T) =CCM 7680(T)).

A tangerine-coloured, Gram-positive actinobacterial strain, designated F10(T), was isolated from the abdominal epidermis of a sea cucumber, Holothuria edulis, collected in seawater off the coast of Japan. A 16S rRNA gene sequence analysis indicated that strain F10(T) was a member of the class Actinobacteria and was most closely related to Nitriliruptor alkaliphilus ANL-iso2(T) (87.4 % sequence similarity). Phylogenetic analyses showed that strain F10(T) represented a novel, deep-rooted, and distinct phylogenetic lineage within the class Actinobacteria and clustered with N. alkaliphilus and uncultured bacteria. The organism had meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and rhamnose and galactose as the diagnostic cell-wall sugars. Strain F10(T) contained C₁₆ :₀ω7c, C₁₆:₀ and C₁₇:₁ω8c as the major cellular fatty acids. The predominant isoprenoid quinone was MK-9 (H₄).The G+C content of the DNA was 68.3 mol%. Based on data from the current polyphasic study, it is proposed that the new marine isolate be placed in a novel genus and be considered a novel species designated Euzebya tangerina gen. nov., sp. nov. within the new family, order and subclass Euzebyaceae fam. nov., Euzebyales ord. nov. and Nitriliruptoridae subclassis nov. in the class Actinobacteria. The type strain of Euzebya tangerina is F10(T) (=NBRC 105439(T) =KCTC 19736(T)).

The bioflocculant ZS-7 was purified to homogeneity by ethanol precipitation, dialysis and gel permeation chromatography (GPC). About 0.98 g of the purified bioflocculant could be recovered from 1L of fermentation broth. The purified bioflocculant was identified as a glycoprotein consisting of polysaccharide (91.5%) and protein (8.4%), with an approximate molecular weight of 6.89 x 10(4). The major component of ZS-7 is an acid polysaccharide including uronic (16.4%), pyruvic (7.1%) and acetic acids (0.5%). It consists of galactose, glucose, mannose and rhamnose in a molar ratio of 142 :2.2 : 4.5 : 3.4. X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) and NMR spectrum of the bioflocculant indicate the presence of carboxyl, hydroxyl, amide, amino, methoxyl and sulfate groups.

A new HPLC-UV technique for the separation and analysis of 10 monosaccharides achieved within 13.5 min using 1-phenyl-3-methyl-5-pyrazolone (PMP) as the labelling molecule of the reductive monosaccharides has been established by combining common high performance liquid chromatography-UV and C18 column. The established technique was applied to the quantification of the monosaccharide components in extract of Silybum marianum. The results showed that the tested 10 monosaccharides as PMP derivatives were baseline separated under the HPLC conditions proposed. It was confirmed that Silybum marianum extract was composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose and arabinose with the molar ratio of 0.66:0.84:0.58:1.0:1.6:0.69:2.7:4.8. Quantitative recoveries of the compositional monosaccharides separated from the extract were in the range of 92.4%-104.0%, and the RSD values fell within 0.68%-3.81%. The results demonstrated that the proposed HPLC method was simple, rapid, convenient, and precise, and it was applicable to the analysis of the compositional monosaccharides of Silybum marianum extract.

Mucilage is thought to play a role in salinity tolerance in certain halophytic species by regulating water ascent and ion transport. The localization and composition of mucilage in the halophyte Kosteletzkya virginica was therefore investigated. Plants were grown in a hydroponic system in the presence or absence of 100mM NaCl and regularly harvested for growth parameter assessment and mucilage analysis with the gas liquid chromatography method. NaCl treatment stimulated shoot growth and biomass accumulation, had little effect on shoot and root water content, and reduced leaf water potential (Psi(w)), osmotic potential (Psi(s)) as well as stomatal conductance (g(s)). Mucilage increased in shoot, stems and roots in response to salt stress. Furthermore, changes were also observed in neutral monosaccharide components. Levels of rhamnose and uronic acid increased with salinity. Staining with a 0.5% alcian blue solution revealed the presence of mucopolyssacharides in xylem vessels and salt-induced mucilaginous precipitates on the leaf abaxial surface. Determination of ion concentrations showed that a significant increase of Na(+) and a decrease of K(+) and Ca(2+) simultaneously occurred in tissues and in mucilage under salt stress. Considering the high proportion of rhamnose and uronic acid in stem mucilage, we suggest that the pectic polysaccharide could be involved in Na(+) fixation, though only a minor fraction of accumulated sodium appeared to be firmly bound to mucilage., (Copyright 2009 Elsevier GmbH. All rights reserved.)

Objective: To study the structure of the polysaccharides constituents in the fruits of Siraitia grosvenorii (Swingle) C. Jeffrey., Methods: SGPS2 was purified by DEAE-cellulose and Sephadex G-200 column chromatography, the component and structure of SGPS2 were analyzed on the basis of spectral and chemical studies by HPLC, IR analysis, partial hydrolysis with acid, methylation analysis, GC and 13C-NMR., Results: The molecular weight of SGPS2 was 650 000. Polysaccharide was composed of L- made up of (1 --> 2, 4) linked rhamnose, (1 --> 4) linked rhamnose residues in main chain; and (1 --> 2) linked rhamnose, (1 --> 3) linked rhamnose in side chains. Terminal residues attached main chain to rhamnose., Conclusion: SGPS2 was composed of rhamnose and curonic acid.

In many Gram-negative bacterial species, rough strains producing truncated lipopolysaccharide (LPS) generally exhibit defects in motility compared with smooth strains. However, the role that LPS plays in bacterial motility is not well understood. The goal of this study was to examine the relationship between LPS defects and motility of Pseudomonas aeruginosa. P. aeruginosa wild-type strain PAO1 and three isogenic mutants with defects in the rmlC, migA and wapR genes and producing truncated core oligosaccharide were investigated in terms of motility, attachment to glass and flagella expression. Compared with the wild-type, the three mutants showed significant retardation in both swarming motility on 0.5 % soft-agar plates and swimming motility on 0.3 % soft-agar plates. Moreover, attachment to abiotic surfaces was observed to be stronger in these mutants. The assembly of flagella appeared to be intact in these strains and the ability of individual cells to swim was unaffected. Flagellin proteins prepared from mutants rmlC and rmd, defective in the production of TDP-l-rhamnose and GDP-d-rhamnose, respectively, were compared and a change in molecular mass was observed only in the rmlC mutant. These data indicated that l-rhamnose, and not its enantiomer, d-rhamnose, is incorporated into the flagellin glycan of P. aeruginosa PAO1. The nucleotide-activated sugar precursor TDP-l-rhamnose is therefore shared between LPS biosynthesis and flagellin glycosylation in P. aeruginosa PAO1. Our results suggest that although biochemical precursors are shared by LPS and flagellin glycan biosynthesis, LPS truncations probably alter flagella-mediated motility in P. aeruginosa by modulating cell-surface attachment but not flagella synthesis.

Enzyme-resistant pectin or modified hairy regions were subjected to size exclusion (HPSEC) and weak anion exchange (WAX) chromatography. Fractions collected after separation were tested for the presence of different pectic epitopes using the monoclonal antibodies LM2, LM5, LM6, and JIM7. Separation by HPSEC showed that based on molecular weight the different epitopes were restricted to distinct molecular weight populations. WAX chromatography resulted in an even better separation of the different pectic epitopes present. A clear separation between arabino galactan type II epitopes and the RG I side chains, (1,5)-alpha-l-arabinan and (1,4)-beta-d-galactan, could be established. Arabinogalactan type II was found in the first populations eluting off the WAX column. The observations made within the ELISA assays of the collected fractions could be confirmed by determination of the sugar composition of the individual populations obtained. The sugar composition of the AGII positive populations eluting off the WAX column shows the presence of significant amounts of rhamnose and galacturonic acid. Together with the delay on an anion exchanger, this observation indicates a possible linkage between RGI and AGII. The volume of the individual fractions collected provides enough material for a maximum of 20 different antibodies to be tested from one analytical separation.

The okra plant, Abelmoschus esculentus (L.) Moench, a native plant from Africa, is now cultivated in many other areas such as Asia, Africa, Middle East, and the southern states of the USA. Okra pods are used as vegetables and as traditional medicines. Sequential extraction showed that the Hot Buffer Soluble Solids (HBSS) extract of okra consists of highly branched rhamnogalacturonan (RG) I containing high levels of acetyl groups and short galactose side chains. In contrast, the CHelating agent Soluble Solids (CHSS) extract contained pectin with less RG I regions and slightly longer galactose side chains. Both pectic populations were incubated with homogeneous and well characterized rhamnogalacturonan hydrolase (RGH), endo-polygalacturonase (PG), and endo-galactanase (endo-Gal), monitoring both high and low molecular weight fragments. RGH is able to degrade saponified HBSS and, to some extent, also non-saponified HBSS, while PG and endo-Gal are hardly able to degrade either HBSS or saponified HBSS. In contrast, PG is successful in degrading CHSS, while RGH and endo-Gal are hardly able to degrade the CHSS structure. These results point to a much higher homogalacturonan (HG) ratio for CHSS when compared to HBSS. In addition, the CHSS contained slightly longer galactan side chains within its RG I region than HBSS. Matrix-assisted laser desorption ionization-time of flight mass spectrometry indicated the presence of acetylated RG oligomers in the HBSS and CHSS enzyme digests and electron spray ionization-ion trap-mass spectrum showed that not only galacturonosyl residues but also rhamnosyl residues in RG I oligomers were O-acetylated. NMR spectroscopy showed that all rhamnose residues in a 20kDa HBSS population were O-acetylated at position O-3. Surprisingly, the NMR data also showed that terminal alpha-linked galactosyl groups were present as neutral side chain substituents. Taken together, these results demonstrate that okra contained RG I structures which have not been reported before for pectic RG I.

Lipopolysaccharide (LPS) is the first defense against changing environmental factors for many bacteria. Here, we report the first structure of the LPS from cyanobacteria based on two strains of marine Synechococcus, WH8102 and CC9311. While enteric LPS contains some of the most complex carbohydrate residues in nature, the full-length versions of these cyanobacterial LPSs have neither heptose nor 3-deoxy-D-manno-octulosonic acid (Kdo) but instead 4-linked glucose as their main saccharide component, with low levels of glucosamine and galacturonic acid also present. Matrix-assisted laser desorption ionization mass spectrometry of the intact minimal core LPS reveals triacylated and tetraacylated structures having a heterogeneous mix of both hydroxylated and nonhydroxylated fatty acids connected to the diglucosamine backbone and a predominantly glucose outer core-like region for both strains. WH8102 incorporated rhamnose in this region as well, contributing to differences in sugar composition and possibly nutritional differences between the strains. In contrast to enteric lipid A, which can be liberated from LPS by mild acid hydrolysis, lipid A from these organisms could be produced by only two novel procedures: triethylamine-assisted periodate oxidation and acetolysis. The lipid A contains odd-chain hydroxylated fatty acids, lacks phosphate, and contains a single galacturonic acid. The LPS lacks any limulus amoebocyte lysate gelation activity. The highly simplified nature of LPSs from these organisms leads us to believe that they may represent either a primordial structure or an adaptation to the relatively higher salt and potentially growth-limiting phosphate levels in marine environments.

Objective: To establish precolumn derivation HPLC method to determine the monosaccharides in Heterosmilax japonica polysaccharide., Methods: Heterosmilax japonica sample was hydrolyzed with trifluoroacetic (TFA)and derivated by 1-phenyl-3-methyl-5-pyrazolone (PMP). The monosaccharide composition of Heterosmilax japonica polysaccharide was carried out by reversed-phase technique on a Kromasil C18 column with a mobile phase composed of 100 mmol/L ammonium acetate buffer (pH5.5) and acetonitrile in the ratio of 77: 23. The detection was carried out at 245 nm., Results: The Heterosmilax japonica polysaccharide composed of mannose, rhamnose, glucose, galactose and xylose with amolar ratio of 66: 1:7590: 452: 528., Conclusion: The HPLC method with precolumn derivatization is appropriate for the analysis of monosaccharide composition of Heterosmilax japonica polysaccharide and the method is simple, quick and accurate.