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Sample records for microbial extracellular polysaccharides

  1. Iron oxyhydroxide mineralization on microbial extracellular polysaccharides

    Energy Technology Data Exchange (ETDEWEB)

    Chan, Clara S.; Fakra, Sirine C.; Edwards, David C.; Emerson, David; Banfield, Jillian F.

    2010-06-22

    Iron biominerals can form in neutral pH microaerophilic environments where microbes both catalyze iron oxidation and create polymers that localize mineral precipitation. In order to classify the microbial polymers that influence FeOOH mineralogy, we studied the organic and mineral components of biominerals using scanning transmission X-ray microscopy (STXM), micro X-ray fluorescence ({mu}XRF) microscopy, and high-resolution transmission electron microscopy (HRTEM). We focused on iron microbial mat samples from a creek and abandoned mine; these samples are dominated by iron oxyhydroxide-coated structures with sheath, stalk, and filament morphologies. In addition, we characterized the mineralized products of an iron-oxidizing, stalk-forming bacterial culture isolated from the mine. In both natural and cultured samples, microbial polymers were found to be acidic polysaccharides with carboxyl functional groups, strongly spatially correlated with iron oxyhydroxide distribution patterns. Organic fibrils collect FeOOH and control its recrystallization, in some cases resulting in oriented crystals with high aspect ratios. The impact of polymers is particularly pronounced as the materials age. Synthesis experiments designed to mimic the biomineralization processes show that the polysaccharide carboxyl groups bind dissolved iron strongly but release it as mineralization proceeds. Our results suggest that carboxyl groups of acidic polysaccharides are produced by different microorganisms to create a wide range of iron oxyhydroxide biomineral structures. The intimate and potentially long-term association controls the crystal growth, phase, and reactivity of iron oxyhydroxide nanoparticles in natural systems.

  2. Microbial short-chain fatty acid production and extracellular enzymes activities during in vitro fermentation of polysaccharides from the seeds of Plantago asiatica L. treated with microwave irradiation.

    Science.gov (United States)

    Hu, Jie-Lun; Nie, Shao-Ping; Li, Chang; Fu, Zhi-Hong; Xie, Ming-Yong

    2013-06-26

    Effects of microwave irradiation on microbial short-chain fatty acid production and the activites of extracellular enzymes during in vitro fermentation of the polysaccharide from Plantago asiatica L. were investigated in this study. It was found that the apparent viscosity, average molecular weight, and particle size of the polysaccharide decreased after microwave irradiation. Reducing sugar amount increased with molecular weight decrease, suggesting the degradation may derive from glycosidic bond rupture. The polysaccharide surface topography was changed from large flakelike structure to smaller chips. FT-IR showed that microwave irradiation did not alter the primary functional groups in the polysaccharide. However, short-chain fatty acid productions of the polysaccharide during in vitro fermentation significantly increased after microwave irradiation. Activities of microbial extracellular enzymes xylanase, arabinofuranosidase, xylosidase, and glucuronidase in fermentation cultures supplemented with microwave irradiation treated polysaccharide were also generally higher than those of untreated polysaccharide. This showed that microwave irradiation could be a promising degradation method for the production of value-added polysaccharides.

  3. Microbially produced extracellular poly-saccharidic Pu(IV)- binding ligands

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    Hung, C.C.; Roberts, K.A.; Schwehr, K.A.; Santschi, P.H. [Texas A and M University at Galveston, 5007 Ave U, Galveston, TX 77551 (United States)

    2005-07-01

    Full text of publication follows: The investigation of the Pu-binding properties of ligands for diverse extracellular polysaccharides (EPS) is of relevance for the quantitative understanding of colloidal barriers to radionuclide migration. The EPS isolated for this study were from four different bacteria species: a) two aerobic soil bacteria: Shewanella putrefaciens CN32 and Pseudomonas fluorescens Biovar II; and b) one anaerobic bacterium, Clostridium sp. BC1. EPS from these bacteria were isolated through repeated ethanol precipitations. The neutral monosaccharides in the EPS from Pseudomonas florescens Biovar II that were determined by GC-MS consisted of rhamnose, fucose, ribose, arabinose, xylose, mannose, galactose and glucose. The potentially Pu(IV) binding EPS ligands were mainly composed of carboxylic acids and other minor poly-anionic groups, e.g., sulphates and phosphates. Up to 70 % of total carbohydrates were hydrophilic uronic acids, and total carbohydrates made up 23-31% of organic carbon for P. florescens Biovar II and 9-17% of organic carbon for S. putrefaciens CN32. Besides the neutral and acidic sugars in the EPS, there were also 2-13 % of more hydrophobic proteins among these bacterial EPS. Pu binding to these exo-polymers showed log Kd values of about 5 - 6, with results strongly dependent on procedural details (e.g., removal of colloids in Pu(IV) tracer and reagent solutions). We hypothesize that the relative hydrophobicity of the EPS ligands affects the outcome in ternary sorption studies with colloidal silica. Experiments with varying relative hydrophobicities of EPS will elucidate the different sorption strengths and/or attachment potentials of the Pu-binding ligands to inorganic surfaces. (authors)

  4. Bacterial Extracellular Polysaccharides Involved in Biofilm Formation

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    Elena P. Ivanova

    2009-07-01

    Full Text Available Extracellular polymeric substances (EPS produced by microorganisms are a complex mixture of biopolymers primarily consisting of polysaccharides, as well as proteins, nucleic acids, lipids and humic substances. EPS make up the intercellular space of microbial aggregates and form the structure and architecture of the biofilm matrix. The key functions of EPS comprise the mediation of the initial attachment of cells to different substrata and protection against environmental stress and dehydration. The aim of this review is to present a summary of the current status of the research into the role of EPS in bacterial attachment followed by biofilm formation. The latter has a profound impact on an array of biomedical, biotechnology and industrial fields including pharmaceutical and surgical applications, food engineering, bioremediation and biohydrometallurgy. The diverse structural variations of EPS produced by bacteria of different taxonomic lineages, together with examples of biotechnological applications, are discussed. Finally, a range of novel techniques that can be used in studies involving biofilm-specific polysaccharides is discussed.

  5. EXTRACELLULAR POLYSACCHARIDES OF POTATO RING ROT PATHOGEN

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    Shafikova Т.N.

    2006-03-01

    Full Text Available Many bacteria, including phytopathogenic ones produce extracellular polysaccharides or exopolysaccharides which are universal molecules. Causal agent of potato ring rot, Clavibacter michiganensis subspecies sepedonicus, secretes exopolysaccharides which role in pathogenesis is poorly investigated. The aim of our research is to ascertain the composition and structure of Clavibacter michiganensis subspecies sepedonicus exopolysaccharides. Exopolysaccharides of Clavibacter michiganensis subspecies sepedonicus are determined to consist of 4-6 anionic and neutral components which have molecular weights from 700 kDa. Glucose is a major monomer of polysaccharides and arabinose, rhamnose and mannose are minor monomers. Glucose is present in α-Dglucopyranose and β-D-glucopyranose configurations. Calcium is determined to be a component of exopolysaccharides. Components of exopolysaccharides of potato ring rot pathogen are probably capableto associate via calcium ions and other ionic interactions that may result in a change of their physiological activity. Further studies of Clavibacter michiganensis subspecies sepedonicus exopolysaccharides composition and structure can serve a base for the synthesis of their chemical analogues with elicitor action.

  6. Biotechnological Aspects of Microbial Extracellular Electron Transfer

    Science.gov (United States)

    Kato, Souichiro

    2015-01-01

    Extracellular electron transfer (EET) is a type of microbial respiration that enables electron transfer between microbial cells and extracellular solid materials, including naturally-occurring metal compounds and artificial electrodes. Microorganisms harboring EET abilities have received considerable attention for their various biotechnological applications, in addition to their contribution to global energy and material cycles. In this review, current knowledge on microbial EET and its application to diverse biotechnologies, including the bioremediation of toxic metals, recovery of useful metals, biocorrosion, and microbial electrochemical systems (microbial fuel cells and microbial electrosynthesis), were introduced. Two potential biotechnologies based on microbial EET, namely the electrochemical control of microbial metabolism and electrochemical stimulation of microbial symbiotic reactions (electric syntrophy), were also discussed. PMID:26004795

  7. Immunological and biochemical characterization of extracellular polysaccharides of mucoralean moulds.

    NARCIS (Netherlands)

    Ruiter, de G.A.

    1993-01-01

    In this thesis the characterization is described of the antigenic determinants (epitopes) of the extracellular polysaccharides (EPSs) from moulds belonging to the order of Mucorales. Detailed knowledge of the structure of these epitopes allows for further development of a new generation of methods f

  8. Extracellular DNA in oral microbial biofilms.

    Science.gov (United States)

    Jakubovics, Nicholas S; Burgess, J Grant

    2015-07-01

    The extracellular matrix of microbial biofilms is critical for surface adhesion and nutrient homeostasis. Evidence is accumulating that extracellular DNA plays a number of important roles in biofilm integrity and formation on hard and soft tissues in the oral cavity. Here, we summarise recent developments in the field and consider the potential of targeting DNA for oral biofilm control.

  9. Inhibition of Streptococcus mutans biofilm formation, extracellular polysaccharide production, and virulence by an oxazole derivative.

    Science.gov (United States)

    Chen, Lulu; Ren, Zhi; Zhou, Xuedong; Zeng, Jumei; Zou, Jing; Li, Yuqing

    2016-01-01

    Dental caries, a biofilm-related oral disease, is a result of disruption of the microbial ecological balance in the oral environment. Streptococcus mutans, which is one of the primary cariogenic bacteria, produces glucosyltransferases (Gtfs) that synthesize extracellular polysaccharides (EPSs). The EPSs, especially water-insoluble glucans, contribute to the formation of dental plaque, biofilm stability, and structural integrity, by allowing bacteria to adhere to tooth surfaces and supplying the bacteria with protection against noxious stimuli and other environmental attacks. The identification of novel alternatives that selectively inhibit cariogenic organisms without suppressing oral microbial residents is required. The goal of the current study is to investigate the influence of an oxazole derivative on S. mutans biofilm formation and the development of dental caries in rats, given that oxazole and its derivatives often exhibit extensive and pharmacologically important biological activities. Our data shows that one particular oxazole derivative, named 5H6, inhibited the formation of S. mutans biofilms and prevented synthesis of extracellular polysaccharides by antagonizing Gtfs in vitro, without affecting the growth of the bacteria. In addition, topical applications with the inhibitor resulted in diminished incidence and severity of both smooth and sulcal surface caries in vivo with a lower percentage of S. mutans in the animals' dental plaque compared to the control group (P < 0.05). Our results showed that this oxazole derivative has the capacity to inhibit biofilm formation and cariogenicity of S. mutans.

  10. Pivotal roles of the outer membrane polysaccharide export and polysaccharide copolymerase protein families in export of extracellular polysaccharides in gram-negative bacteria.

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    Cuthbertson, Leslie; Mainprize, Iain L; Naismith, James H; Whitfield, Chris

    2009-03-01

    Many bacteria export extracellular polysaccharides (EPS) and capsular polysaccharides (CPS). These polymers exhibit remarkably diverse structures and play important roles in the biology of free-living, commensal, and pathogenic bacteria. EPS and CPS production represents a major challenge because these high-molecular-weight hydrophilic polymers must be assembled and exported in a process spanning the envelope, without compromising the essential barrier properties of the envelope. Emerging evidence points to the existence of molecular scaffolds that perform these critical polymer-trafficking functions. Two major pathways with different polymer biosynthesis strategies are involved in the assembly of most EPS/CPS: the Wzy-dependent and ATP-binding cassette (ABC) transporter-dependent pathways. They converge in an outer membrane export step mediated by a member of the outer membrane auxiliary (OMA) protein family. OMA proteins form outer membrane efflux channels for the polymers, and here we propose the revised name outer membrane polysaccharide export (OPX) proteins. Proteins in the polysaccharide copolymerase (PCP) family have been implicated in several aspects of polymer biogenesis, but there is unequivocal evidence for some systems that PCP and OPX proteins interact to form a trans-envelope scaffold for polymer export. Understanding of the precise functions of the OPX and PCP proteins has been advanced by recent findings from biochemistry and structural biology approaches and by parallel studies of other macromolecular trafficking events. Phylogenetic analyses reported here also contribute important new insight into the distribution, structural relationships, and function of the OPX and PCP proteins. This review is intended as an update on progress in this important area of microbial cell biology.

  11. Incorporation of bacterial extracellular polysaccharide by black fly larvae (Simuliidae)

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    Couch, C.A.; Meyer, J.L.; Hall, R.O.

    1996-01-01

    Black fly larvae (Simulium) assimilated, with high efficiency (80-90%), bacterial extracellular polysaccharide (EPS) extracted from laboratory cultures of a pseudomonad isolated from the Ogeechee River. Incorporation was traced using 13C-labelled EPS offered to larvae as a coating on a mixture of 1-??m latex beads and kaolin particles. These EPS-coated particles were used to simulate natural particles, both living and dead. Solubility, protein, and nitrogen content of the EPS suggested it was a slime rather than a capsular polysaccharide. Glycosyl composition of the EPS was glucose and galactose in ?? and ?? linkages, with pyruvate, succinate, and possibly malonate constituent groups. To evaluate the incorporation of C derived from protein associated with the EPS matrix, feeding experiments were conducted using EPS with and without proteins extracted. Black fly larvae incorporated 7.2 ??g EPS C larva-1 d-1 from EPS that did not have proteins extracted, and 19.5 ??g EPS C larva-1 d-1 from EPS with proteins extracted. Carbon in protein that is typically associated with EPS was not solely or selectively incorporated. EPS incorporation rates are similar to rates of cellular bacterial carbon incorporation previously estimated for Ogeechee River black fly larvae. If EPS is generally available as a food resource, the importance of bacteria in detrital food webs may be underestimated by studies that examine only the consumption of bacterial cells.

  12. High activity and low temperature optima of extracellular enzymes in Arctic sediments: implications for carbon cycling by heterotrophic microbial communities

    DEFF Research Database (Denmark)

    Arnosti, C.; Jørgensen, BB

    2003-01-01

    The rate of the initial step in microbial remineralization of organic carbon, extracellular enzymatic hydrolysis, was investigated as a function of temperature in permanently cold sediments from 2 fjords on the west coast of Svalbard (Arctic Ocean). We used 4 structurally distinct polysaccharides...... with recent studies of psychrophilic sulfate reducers isolated from Svalbard sediments. A calculation of potential carbon flow into the microbial food chain demonstrated that the activity of just one type of polysaccharide-hydrolyzing enzyme could in theory supply 21 to 100% of the carbon consumed via sulfate...... reduction across the temperature range investigated here. These characteristics suggest that these extracellular enzymes are well adapted to permanently cold temperatures....

  13. Extracellular acidic polysaccharide production by a two-membered bacterial coculture.

    Science.gov (United States)

    Kurata, Shinya; Yamada, Kazutaka; Takatsu, Kyoko; Hanada, Satoshi; Koyama, Osamu; Yokomaku, Toyokazu; Kamagata, Yoichi; Kanagawa, Takahiro; Kurane, Ryuichiro

    2003-01-01

    A two-membered coculture of strains KYM-7 and KYM-8, identified as Cellulomonas cellulans and Agrobacterium tumefaciens, respectively, produced a large amount of an extracellular polysaccharide, designated APK-78, from starch. Each strain in pure culture produced only very little amount of polysaccharide from starch; the coexistence of the two strains from the early stage of cultivation was indispensable for a large amount of polysaccharide to be produced. The polysaccharide APK-78 was acidic and composed of glucose, galactose, succinic acid, and pyruvic acid with a molar ratio of 8.1:1.0:1.7:1.0, indicating that it is a succinoglycan type of polysaccharide.

  14. Microbial Extracellular Polymeric Substances (EPSs in Ocean Systems

    Directory of Open Access Journals (Sweden)

    Alan W. Decho

    2017-05-01

    Full Text Available Microbial cells (i.e., bacteria, archaea, microeukaryotes in oceans secrete a diverse array of large molecules, collectively called extracellular polymeric substances (EPSs or simply exopolymers. These secretions facilitate attachment to surfaces that lead to the formation of structured ‘biofilm’ communities. In open-water environments, they also lead to formation of organic colloids, and larger aggregations of cells, called ‘marine snow.’ Secretion of EPS is now recognized as a fundamental microbial adaptation, occurring under many environmental conditions, and one that influences many ocean processes. This relatively recent realization has revolutionized our understanding of microbial impacts on ocean systems. EPS occur in a range of molecular sizes, conformations and physical/chemical properties, and polysaccharides, proteins, lipids, and even nucleic acids are actively secreted components. Interestingly, however, the physical ultrastructure of how individual EPS interact with each other is poorly understood. Together, the EPS matrix molecules form a three-dimensional architecture from which cells may localize extracellular activities and conduct cooperative/antagonistic interactions that cannot be accomplished efficiently by free-living cells. EPS alter optical signatures of sediments and seawater, and are involved in biogeomineral precipitation and the construction of microbial macrostructures, and horizontal-transfers of genetic information. In the water-column, they contribute to the formation of marine snow, transparent exopolymer particles (TEPs, sea-surface microlayer biofilm, and marine oil snow. Excessive production of EPS occurs during later-stages of phytoplankton blooms as an excess metabolic by product and releases a carbon pool that transitions among dissolved-, colloidal-, and gel-states. Some EPS are highly labile carbon forms, while other forms appear quite refractory to degradation. Emerging studies suggest that EPS

  15. Modulation of surgical fibrosis by microbial zwitterionic polysaccharides

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    Ruiz-Perez, Begonia; Chung, Doo R.; Sharpe, Arlene H.; Yagita, Hideo; Kalka-Moll, Wiltrud M.; Sayegh, Mohamed H.; Kasper, Dennis L.; Tzianabos, Arthur O.

    2005-11-01

    Bacterial carbohydrates have long been considered T cell-independent antigens that primarily induce humoral immune responses. Recently, it has been demonstrated that bacterial capsules that possess a zwitterionic charge motif can activate CD4+ T cells after processing and presentation by antigen-presenting cells. Here we show that these zwitterionic polysaccharides can prevent T helper 1-mediated fibrosis by signaling for the release of IL-10 from CD4+ T cells in vivo. IL-10 production by these T cells and their ability to prevent fibrosis is controlled by the inducible costimulator (ICOS)-ICOS ligand pathway. These data demonstrate that the interaction of the zwitterionic polysaccharides with T cells results in modulation of surgical fibrosis in vivo and suggest a previously undescribed approach to "harnessing" T cell function to prevent inflammatory tissue disorders in humans. IL-10 | microbial polysaccharides | inducible costimulator

  16. Extracellular Polysaccharides Matrix - An Often Forgotten Virulence Factor in Oral Biofilm Research

    Institute of Scientific and Technical Information of China (English)

    Hyun Koo; Jin Xiao; Marlise I. Klein

    2009-01-01

    @@ Oral diseases related to dental biofilms continue to afflict the majority of the world's population. Among them, dental caries continues to be the single most prevalent and costly oral infectious disease (Marsh, 2003; Dye et al., 2007). Dental caries results from the interaction of specific bacteria with constituents of the diet within a dental biofilm known as plaque (Bowen, 2002). Sucrose is considered to be the "arch criminal" from the dietary aspect because it serves as a substrate for synthesis of extracellular (EPS) and intracellular (IPS) polysaccharides in dental biofilm and is also fermentable (Bowen, 2002). However, it is important to emphasize that additional sugars and starch can certainly contribute to the pathogenesis (Bowen et al., 1980; Firestone et al., 1982; Thurnheer et al., 2008). Streptococcus mutans (S. mutans), a member of the oral microbial community, is generally regarded as the primary microbial culprit although additional microorganisms may be involved (Hamada and Slade, 1980; Loesche, 1986; Beighton, 2005). This bacterium (i) effectively utilizes dietary sucrose (and possibly starch) to synthesize large amounts of EPS through glucosyltransferases (Gtfs) and a fructosyltransferase (Ftfs), (ii) adheres tenaciously to glucan-coated surfaces, and (iii) is also acidogenic and acid-tolerant, which are critical virulence properties involved in the pathogenesis of dental caries.

  17. Effect of exogenous extracellular polysaccharides on the desiccation and freezing tolerance of rock-inhabiting phototrophic microorganisms.

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    Knowles, Emily J; Castenholz, Richard W

    2008-11-01

    Two major stresses that threaten rock-inhabiting microbial communities are desiccation and freezing; both result in a loss of liquid water in the cells. The mechanisms necessary to tolerate these extremes may be similar, but are not well understood. In both cases extracellular polysaccharides (EPS) seem to play an important role. This study examines whether the EPS released by a rock-inhabiting phototroph can have a protective effect on other members of similar and neighboring microbial communities. This interaction was modeled by adding EPS isolated from the cryptoendolithic cyanobacterium Nostoc sp. to cells of the cryptoendolithic green alga Chlorella sp. and to cells of the epilithic cyanobacterium Chroococcidiopsis sp. The cells were then subjected to desiccation and freezing and the survival rates were determined by vital staining, using membrane integrity as a measure of viability. The results clearly demonstrate the importance of exogenous EPS in the desiccation tolerance of both species, while mixed results were found for the freezing trials.

  18. Results of the Study of Mutagenic Effects of Microbial Polysaccharides

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    Natalya A. Sidorova

    2016-06-01

    Full Text Available The article presents the results of a study of mutagenic effects of Pseudomonas alcaligenеs polysaccharides. Pseudomonas genus – non-fermentative ubiquitous bacteria, having specific metabolic cycles and unique physical, chemical and biological properties was used as a producer of natural exopolysaccharides. In an experiment using the Ames test, three variants of test compounds were studied: 1. a compound of the Pseudomonas alcaligenes biofilm, 2. exopolysaccharide matrix and the microorganism cell wall compound, and 3. actually the microbial exopolysaccharide. In all cases the lack of mutagen action of polysaccharides of Pseudomonas alcaligenes is proved that make them perspective for use as nanomaterials of new generation – alternative wound coverings.

  19. Structural investigation of an extracellular polysaccharide produced by the cariogenic bacterium Streptococcus mutans strain UA159

    NARCIS (Netherlands)

    Li, Bo; Dobruchowska, Justyna M.; Hoogenkamp, Michel A.; Gerwig, Gerrit J.

    2012-01-01

    The structure of an extracellular polysaccharide EPS159 produced from sucrose by Streptococcus mutans UA159 was investigated through the main oligosaccharides obtained from partial acid hydrolysis, monosaccharide/methylation analysis, and 1D/2D H-1 NMR spectroscopy. The results showed that EPS159 co

  20. Extracellular polysaccharides as target compounds for the immunological detection of Aspergillus and Penicillium in food.

    NARCIS (Netherlands)

    Kamphuis, H.J.

    1992-01-01

    This thesis is devoted to the immunological detection of Aspergillus and Penicillium in food products. More specifically, the immunogenicity, antigenicity, production and structure of the water-soluble extracellular polysaccharides (EPS) of these moulds have been studied, and a latex-agglutination a

  1. Structure of an extracellular polysaccharide produced by Lactobacillus rhamnosus strain C83

    Energy Technology Data Exchange (ETDEWEB)

    Vanhaverbeke, C.; Bosso, C.; Colin-Morel, P.; Gey, C.; Heyraud, A. [Centre de Recherches sur les Macromolecules Vegetales, CNRS and Universite Joseph Fourier, B.P.53, F-38041 Grenoble (France); Gamar-Nourani, L.; Blondeau, K.; Simonet, J.-M. [Institut de Genetique et Microbiologie, Laboratoire de Genetique Moleculaire des Bacteries d' Interet Industriel, CNRS URA 2225, Batiment 360, Universite de Paris Sud, F-91405 Orsay (France)

    1998-12-31

    The extracellular polysaccharide produced by Lactobacillus rhamnosus strain C83 was found to be composed of d-glucose and d-galactose in a molar ratio of 2:3. The primary structure of the polysaccharide was shown by sugar analysis, methylation analysis, FABMS, partial acid hydrolysis and nuclear magnetic resonance (NMR) spectroscopy to consist of a pentasaccharide repeating unit having the following structure:-3)-{alpha}-d-Glcp-(1-2)-{beta}-d-Galf-(1-6)-{alpha}-d-Galp-(1-6) -{alpha}-d-Glcp-(1-3)-{beta}-d-Galf-(1-. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

  2. Extracellular enzyme activity assay as indicator of soil microbial functional diversity and activity

    DEFF Research Database (Denmark)

    Hendriksen, Niels Bohse; Winding, Anne

    2012-01-01

    of soil microbial functions is still needed. In soil, enzymes originate from a variety of organisms, notably fungi and bacteria and especially hydrolytic extracellular enzymes are of pivotal importance for decomposition of organic substrates and biogeochemical cycling. Their activity will reflect...... the functional diversity and activity of the microorganisms involved in decomposition processes. Their activity has been measured by the use of fluorogenic model substrates e.g. methylumbelliferyl (MUF) substrates for a number of enzymes involved in the degradation of polysacharides as cellulose, hemicellulose...... and chitin, while degradation of proteins has been followed by amino-methyl-coumaric substrates (AMC). Based on these fluorogenic substrates the Extracellular Enzyme Activity assay was optimized as a microwell based standardized assay for the activity of enzymes involved in degradation of polysaccharides...

  3. Extracellular polysaccharides are involved in the attachment of Azospirillum brasilense and Rhizobium leguminosarum to arbuscular mycorrhizal structures

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    V Bianciotto

    2009-12-01

    Full Text Available Arbuscular mycorrhizal (AM fungi, one of the most important component of the soil microbial community, establish physical interactions with naturally occurring and genetically modified bacterial biofertilizers and biopesticides, commonly referred to as plant growth-promoting rhizobacteria (PGPR. We have used a genetic approach to investigate the bacterial components possibly involved in the attachment of two PGPR (Azospirillum and Rhizobium to AM roots and AM fungal structures. Mutants affected in extracellular polysaccharides (EPS have been tested in in vitro adhesion assays and shown to be strongly impaired in the attachment to both types of surfaces as well as to quartz fibers. Anchoring of rhizobacteria to AM fungal structures may have special ecological and biotechnological significance because it may facilitate colonisation of new rhizospheres by the bacteria, and may be an essential trait for the development of mixed inocula.

  4. Extracellular polysaccharides are involved in the attachment of Azospirillum brasilense and Rhizobium leguminosarum to arbuscular mycorrhizal structures.

    Science.gov (United States)

    Bianciotto, V; Andreotti, S; Balestrini, R; Bonfante, P; Perotto, S

    2001-01-01

    Arbuscular mycorrhizal (AM) fungi, one of the most important component of the soil microbial community, establish physical interactions with naturally occurring and genetically modified bacterial biofertilizers and biopesticides, commonly referred to as plant growth-promoting rhizobacteria (PGPR). We have used a genetic approach to investigate the bacterial components possibly involved in the attachment of two PGPR (Azospirillum and Rhizobium) to AM roots and AM fungal structures. Mutants affected in extracellular polysaccharides (EPS) have been tested in in vitro adhesion assays and shown to be strongly impaired in the attachment to both types of surfaces as well as to quartz fibers. Anchoring of rhizobacteria to AM fungal structures may have special ecological and biotechnological significance because it may facilitate colonisation of new rhizospheres by the bacteria, and may be an essential trait for the development of mixed inocula.

  5. Microbial extracellular electron transfer and its relevance to iron corrosion.

    Science.gov (United States)

    Kato, Souichiro

    2016-03-01

    Extracellular electron transfer (EET) is a microbial metabolism that enables efficient electron transfer between microbial cells and extracellular solid materials. Microorganisms harbouring EET abilities have received considerable attention for their various biotechnological applications, including bioleaching and bioelectrochemical systems. On the other hand, recent research revealed that microbial EET potentially induces corrosion of iron structures. It has been well known that corrosion of iron occurring under anoxic conditions is mostly caused by microbial activities, which is termed as microbiologically influenced corrosion (MIC). Among diverse MIC mechanisms, microbial EET activity that enhances corrosion via direct uptake of electrons from metallic iron, specifically termed as electrical MIC (EMIC), has been regarded as one of the major causative factors. The EMIC-inducing microorganisms initially identified were certain sulfate-reducing bacteria and methanogenic archaea isolated from marine environments. Subsequently, abilities to induce EMIC were also demonstrated in diverse anaerobic microorganisms in freshwater environments and oil fields, including acetogenic bacteria and nitrate-reducing bacteria. Abilities of EET and EMIC are now regarded as microbial traits more widespread among diverse microbial clades than was thought previously. In this review, basic understandings of microbial EET and recent progresses in the EMIC research are introduced.

  6. Effects of light wavelengths on extracellular and capsular polysaccharide production by Nostoc flagelliforme.

    Science.gov (United States)

    Han, Pei-pei; Sun, Ying; Jia, Shi-ru; Zhong, Cheng; Tan, Zhi-lei

    2014-05-25

    The influences of different wavelengths of light (red 660nm, yellow 590nm, green 520nm, blue 460nm, purple 400nm) and white light on extracellular polysaccharide (EPS) and capsular polysaccharide (CPS) production by Nostoc flagelliforme in liquid culture were demonstrated in this study. The results showed that, compared with white light, red and blue lights significantly increased both EPS and CPS production while yellow light reduced their production; purple and green lights stimulated EPS production but inhibited CPS formation. Nine constituent monosaccharides and one uronic acid were detected in both EPS and CPS, and their ratios showed significant differences among treatment with different light wavelengths. However, the advanced structure of EPS and CPS from various light conditions did not present obvious difference through Fourier transform infrared spectroscopy and X-ray diffraction characterization. These findings establish a basis for development of high-yielding polysaccharide production process and understanding their regulation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Extracellular polysaccharide composition of Azospirillum brasilense and its relation with cell aggregation.

    Science.gov (United States)

    Burdman, S; Jurkevitch, E; Soria-Díaz, M E; Serrano, A M; Okon, Y

    2000-08-15

    The exopolysaccharide (EPS) and capsular polysaccharide (CPS) composition of four Azospirillum brasilense strains differing in their aggregation capacity was analyzed by high performance anion exchange chromatography. When growing the different strains in an aggregation inducing medium containing a high carbon:nitrogen (C:N) ratio, both EPS and CPS showed a positive correlation between aggregation and the relative amount of arabinose. Arabinose was not detected in polysaccharides from Sp72002, a pleiotrophic Tn5 mutant strain impaired in aggregation. Arabinose was also not detected in extracellular polysaccharides of bacteria grown in a low C:N ratio, non-inducing aggregation medium, with exception for a relatively small amount found in the CPS of FAJ0204, a super-aggregating mutant strain. The only monosaccharides able to significantly inhibit aggregation at low sugar concentration when tested in a bioassay were arabinose (at a higher extent) and galactose. The possibility that residues of arabinose present in the extracellular polysaccharides are involved in the aggregation of A. brasilense is discussed.

  8. Structure of an Extracellular Polysaccharide from a Strain of Lactic Acid Bacteria

    Institute of Scientific and Technical Information of China (English)

    顾笑梅; 马桂荣; 吴厚铭

    2003-01-01

    A new extracellular polysaccharide (EPS-I) isolated and purified from Z222, a strain of Lactic acid bacteria has been investigated. Sugar composition analysis, methylation analysis and 1H NMR and 13C NMR spectroscopy reveal that the EPS-I is composed of a pentasaccharide repeating unit. The sequence of sugar residue was determined by using two-dlmensional NMR spectroscopy, including heteronudear multiple-bond correlation(HMBC) and nuclear overhauser effect spectroscopy (NOESY).

  9. Extracellular polysaccharides as target compounds for the immunological detection of Aspergillus and Penicillium in food.

    OpenAIRE

    Kamphuis, H.J.

    1992-01-01

    This thesis is devoted to the immunological detection of Aspergillus and Penicillium in food products. More specifically, the immunogenicity, antigenicity, production and structure of the water-soluble extracellular polysaccharides (EPS) of these moulds have been studied, and a latex-agglutination assay, based on the detection of EPS has been developed.For the detection of moulds many methods are available, each of them with specific advantages and disadvantages, mostly related to reliability...

  10. Effect of pH values on the extracellular polysaccharide secreted by Acidithiobacillus ferrooxidans during chalcopyrite bioleaching

    Institute of Scientific and Technical Information of China (English)

    Run-lan Yu; Jing Liu; Jian-xi Tan; Wei-min Zeng; Li-juan Shi; Guo-hua Gu; Wen-qing Qin; Guan-zhou Qiu

    2014-01-01

    The pH value plays an important role in the bioleaching of sulphide minerals. The effect of pH values on the extracellular poly-saccharide secreted by Acidithiobacillus ferrooxidans was investigated in different phases of bacterial growth during chalcopyrite bioleach-ing. It is found that extracellular polysaccharide secretion from the cells attached to chalcopyrite is more efficiently than that of the free cells in the bioleaching solution. Three factors, pH values, the concentration of soluble metal ions, and the bacterial growth and metabolism, affect extracellular polysaccharide secretion in the free cells, and are related to the bacterial growth phase. Extracellular polysaccharide secretion from the attached cells is mainly dependent on the pH value of the bacterial culture.

  11. A Polysaccharide-Degrading Marine Bacterium Flammeovirga sp.MY04 and Its Extracellular Agarase System

    Institute of Scientific and Technical Information of China (English)

    HAN Wenjun; GU Jingyan; YAN Qiujie; LI Jungang; WU Zhihong; GU Qianqun; LI Yuezhong

    2012-01-01

    Bacteria of the genus Flammeovirga can digest complex polysaccharides(CPs),but no details have been reported regarding the CP depolymerases of these bacteria.MY04,an agarolytic marine bacterium isolated from coastal sediments,has been identified as a new member of the genus Flammeovirga.The MY04 strain is able to utilize multiple CPs as a sole carbon source and grows well on agarose,mannan,or xylan.This strain produces high concentrations of extracellular proteins (490mgL-1± 18.2 mgL-1liquid culture)that exhibit efficient and extensive degradation activities on various polysaccharides,especially agarose.These proteins have an activity of 310 U mg-1± 9.6 Umg-1 proteins.The extracellular agarase system(EAS)in the crude extracellular enzymes contains at least four agarose depolymerases,which are with molecular masses of approximately 30-70 kDa.The EAS is stable at a wide range of pH values(6.0-11.0),temperatures(0-50℃),and sodium chloride(NaCl)concentrations(0-0.9mol L-1).Two major degradation products generated from agarose by the EAS are identified to be neoagarotetraose and neoagarohexaose,suggesting that β-agarases are the major constituents of the MY04 EAS.These results suggest that the Flammeovirga strain MY04 and its polysaccharide-degradation system hold great promise in industrial applications.

  12. Imaging hydrated microbial extracellular polymers: Comparative analysis by electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Dohnalkova, A.C.; Marshall, M. J.; Arey, B. W.; Williams, K. H.; Buck, E. C.; Fredrickson, J. K.

    2011-01-01

    Microbe-mineral and -metal interactions represent a major intersection between the biosphere and geosphere but require high-resolution imaging and analytical tools for investigating microscale associations. Electron microscopy has been used extensively for geomicrobial investigations and although used bona fide, the traditional methods of sample preparation do not preserve the native morphology of microbiological components, especially extracellular polymers. Herein, we present a direct comparative analysis of microbial interactions using conventional electron microscopy approaches of imaging at room temperature and a suite of cryogenic electron microscopy methods providing imaging in the close-to-natural hydrated state. In situ, we observed an irreversible transformation of the hydrated bacterial extracellular polymers during the traditional dehydration-based sample preparation that resulted in their collapse into filamentous structures. Dehydration-induced polymer collapse can lead to inaccurate spatial relationships and hence could subsequently affect conclusions regarding nature of interactions between microbial extracellular polymers and their environment.

  13. On the chemical dynamics of extracellular polysaccharides in the high Arctic surface microlayer

    Science.gov (United States)

    Gao, Q.; Leck, C.; Rauschenberg, C.; Matrai, P. A.

    2012-07-01

    The surface microlayer (SML) represents a unique system of which the physicochemical characteristics may differ from those of the underlying subsurface seawater (SSW). Within the Arctic pack ice area, the SML has been characterized as enriched in small colloids of biological origin, resulting from extracellular polymeric secretions (EPS). During the Arctic Summer Cloud Ocean Study (ASCOS) in August 2008, particulate organic matter (POM, with size range > 0.22 μm) and dissolved organic matter (DOM, fucose and glucose. The mean concentrations of total hydrolysable neutral sugars in SSW were 94.9 ± 37.5 nM in high molecular weight (HMW) DOM (> 5 kDa) and 64.4 ± 14.5 nM in POM. The enrichment of polysaccharides in the SML appeared to be a common feature, with EFs ranging from 1.7 to 7.0 for particulate polysaccharides and 3.5 to 12.1 for polysaccharides in the HMW DOM fraction. A calculated monosaccharide yield suggests that polymers in the HMW DOM fraction were scavenged, without substantial degradation, into the SML. Bubble scavenging experiments showed that newly aggregated particles could be formed abiotically by coagulation of low molecular weight nanometer-sized gels. Aerosol particles, artificially generated by bubbling experiments, were enriched in polysaccharides by factors of 22-70, relative to the source seawater. We propose that bubble scavenging of surface-active polysaccharides could be one of the possible mechanisms for the enrichment of polysaccharides in the high Arctic open lead SML.

  14. On the chemical dynamics of extracellular polysaccharides in the high Arctic surface microlayer

    Directory of Open Access Journals (Sweden)

    Q. Gao

    2012-01-01

    Full Text Available The surface microlayer (SML represents a unique system of which the physicochemical characteristics may differ from those of the underlying subsurface seawater (SSW. Within the Arctic pack ice area, the SML has been characterized as enriched in small colloids of biological origin, resulting from extracellular polymeric secretions (EPS. During the Arctic Summer Cloud-Ocean Study (ASCOS in August 2008, particulate and dissolved organic matter (POM, DOM samples were collected and chemically characterized from the SML and the corresponding SSW at an open lead centered at 87.5° N and 5° E. Total organic carbon was persistently enriched in the SML with a mean enrichment factor (EF of 1.45 ± 0.41, whereas sporadic depletions of dissolved carbohydrates and amino acids were observed. Monosaccharide compositional analysis reveals that EPS in the Arctic lead was formed mainly of distinctive heteropolysaccharides, enriched in xylose, fucose and glucose. The mean concentrations of total hydrolysable neutral sugars in SSW were 94.9 ± 37.5 nM in high molecular weight (HMW DOM and 64.4 ± 14.5 nM in POM. The enrichment of polysaccharides in the SML appeared to be a common feature, with EFs ranging from 1.7 to 7.0 for particulate polysaccharides and 3.5 to 12.1 for polysaccharides in the HMW DOM fraction. A calculated monosaccharide yield suggests that polymers in the HMW DOM fraction were scavenged, without substantial degradation, into the SML. Bubble scavenging experiments showed that newly aggregated particles could be formed abiotically by coagulation of low molecular weight nanometer-sized gels. Experimentally-generated aerosol particles were enriched in polysaccharides by factors of 22–70, relative to the source seawater. We propose that bubble scavenging of surface-active polysaccharides was one of the possible mechanisms for the enrichment of polysaccharides in the SML.

  15. Extracellular polysaccharides mediate pilus retraction during social motility of Myxococcus xanthus.

    Science.gov (United States)

    Li, Yinuo; Sun, Hong; Ma, Xiaoyuan; Lu, Ann; Lux, Renate; Zusman, David; Shi, Wenyuan

    2003-04-29

    Myxococcus xanthus is a Gram-negative bacterium with a complex life cycle that includes vegetative swarming and fruiting-body formation. Social (S)-motility (coordinated movement of large cell groups) requires both type IV pili and fibrils (extracellular matrix material consisting of polysaccharides and protein). Little is known about the role of this extracellular matrix, or fibril material, in pilus-dependent motility. In this study, mutants lacking fibril material and, therefore, S-motility were found to be hyperpiliated. We demonstrated that addition of fibril material resulted in pilus retraction and rescued this phenotype. The fibril material was further examined to determine the component(s) that were responsible for triggering pilus retraction. Protein-free fibril material was found to be highly active in correcting hyperpiliation. However, the amine sugars present in hydrolyzed fibril material, e.g., glucosamine and N-acetylglucosamine (GlcNAc) had no effect on fibril(-) mutants, but, interestingly, cause hyperpiliation in wild-type cells. In contrast, chitin, a natural GlcNAc polymer, was found to restore pilus retraction in hyperpiliated mutants, indicating that a polysaccharide containing amine sugars is likely required for pilus retraction. These data suggest that the interaction of type IV pili with amine-containing polysaccharides on cell and slime-trail surfaces may trigger pilus retraction, resulting in S-motility and slime-trailing behaviors.

  16. Characterization of an extracellular polysaccharide produced by a Pseudomonas strain grown on glycerol.

    Science.gov (United States)

    Freitas, Filomena; Alves, Vitor D; Pais, Joana; Costa, Nuno; Oliveira, Cristina; Mafra, Luís; Hilliou, Loic; Oliveira, Rui; Reis, Maria A M

    2009-01-01

    A new extracellular charged polysaccharide composed mainly by galactose, with lower amounts of mannose, glucose and rhamnose, was produced by the cultivation of Pseudomonas oleovorans NRRL B-14682 using glycerol as the sole carbon source. Thermal and solid-state NMR analysis showed that this polymer is essentially amorphous, with a glass transition temperature of 155.7 degrees C. The exopolysaccharide aqueous solutions have viscoelastic properties similar to that of Guar gum, but with affinity to salts as a result of its polyelectrolyte character. In addition, the exopolysaccharide has demonstrated good flocculating and emulsifying properties and film-forming capacity. These properties make this polymer a good alternative to more expensive natural polysaccharides, such as Guar gum, in several applications in the food, pharmaceutical, cosmetic, textile, paper and petroleum industries.

  17. Membrane biofouling by extracellular polymeric substances or soluble microbial products from membrane bioreactor sludge.

    Science.gov (United States)

    Ramesh, A; Lee, D J; Lai, J Y

    2007-03-01

    This study extracted the soluble microbial products and loosely bound and tightly bound extracellular polymeric substances (EPS) from suspended sludge from a membrane bioreactor, original and aerobically/anaerobically digested, and compared their fouling potentials on a microfiltration membrane. The resistance of cake layer accounts for 95-98% of the total filtration resistances when filtering the whole sludges, with anaerobically digested sludge presenting the highest resistance among the three tested sludges. The tightly bound EPS has the highest potential to foul the membrane; however, the loosely bound EPS contribute most of the filtration resistances of the whole sludges. The foulants corresponding to the irreversible fouling have chemical fingerprints similar to those from loosely bound EPS, which have a greater predilection to proteins and humic substances than to polysaccharides.

  18. Sulfation of extracellular polysaccharides of red microalgae: preparation, characterization and properties.

    Science.gov (United States)

    Geresh, Shimona; Mamontov, Anna; Weinstein, Jacob

    2002-01-04

    Polysaccharides are natural polymers with a variety of properties that may be translated into significant commercial applications. A program of chemical modifications of the extracellular polysaccharides of red microalgae, such as Porphyridium sp. and Rhodella reticulata, has been undertaken by our group in order to tailor new properties and hence to broaden the spectrum of potential applications. These algal biopolymers are anionic in nature due to the presence of uronic acids (about 10%) and sulfate half esters (about 7%). In the current study, the sulfate content of these biopolymers was increased to 35-40% by means of sulfation agents such as pyridine SO(3), DMF.SO(3) and ClSO(3)H. Reaction conditions were optimized in a model system based on potato starch as the model polysaccharide (type of reagent, temperature and time of reaction). After work-up procedures, the highest sulfate content was obtained by sulfation of the polysaccharide of Porphyridium sp. with a mixture of ClSO(3)H and pyridine at 70 degrees C for 1 h. The sulfated products were characterized by chemical and rheological analyses, IR spectroscopy, and GPC-HPLC chromatography. "Oversulfated" polymers (having sulfate contents exceeding 20%) with high molecular weights were found to inhibit mammalian cell growth when used at certain concentrations; for example, over 80% inhibition was obtained when oversulfated polymers at a concentration of 200 microg/ml were tested on T-cell lymphoma line 24-1. These preliminary results indicate that the modified polysaccharides do indeed exhibit potential therapeutic properties.

  19. Production and composition of extracellular polysaccharide synthesized by a Rhizobium isolate of Vigna mungo (L.) Hepper.

    Science.gov (United States)

    Mandal, Santi Mohan; Ray, Bimalendu; Dey, Satyahari; Pati, Bikas Ranjan

    2007-08-01

    An extracellular polysaccharide (EPS) was produced by a Rhizobium sp. isolated from the root nodules of Vigna mungo (L.) Hepper. Maximum EPS production (346 mg l(-1)) was when the yeast extract basal medium was supplemented with mannitol (1%), biotin (1.5 mg l(-1)) and asparagine (0.3%). Ribose (53%) and mannose (47%) were the principle monomers of the EPS. Chemical, chromatographic and spectroscopic analysis showed that this polymer, which has Man(4)Rib(1) as an oligomeric subunit, has an apparent molecular mass of 750 kDa.

  20. On the chemical dynamics of extracellular polysaccharides in the high Arctic surface microlayer

    Directory of Open Access Journals (Sweden)

    Q. Gao

    2012-07-01

    Full Text Available The surface microlayer (SML represents a unique system of which the physicochemical characteristics may differ from those of the underlying subsurface seawater (SSW. Within the Arctic pack ice area, the SML has been characterized as enriched in small colloids of biological origin, resulting from extracellular polymeric secretions (EPS. During the Arctic Summer Cloud Ocean Study (ASCOS in August 2008, particulate organic matter (POM, with size range > 0.22 μm and dissolved organic matter (DOM, < 0.22 μm, obtained after filtration samples were collected and chemically characterized from the SML and the corresponding SSW at an open lead centered at 87.5° N and 5° E. Total organic carbon was persistently enriched in the SML with a mean enrichment factor (EF of 1.45 ± 0.41, whereas sporadic depletions of dissolved carbohydrates and amino acids were observed. Monosaccharide compositional analysis reveals that EPS in the Arctic lead was formed mainly of distinctive heteropolysaccharides, enriched in xylose, fucose and glucose. The mean concentrations of total hydrolysable neutral sugars in SSW were 94.9 ± 37.5 nM in high molecular weight (HMW DOM (> 5 kDa and 64.4 ± 14.5 nM in POM. The enrichment of polysaccharides in the SML appeared to be a common feature, with EFs ranging from 1.7 to 7.0 for particulate polysaccharides and 3.5 to 12.1 for polysaccharides in the HMW DOM fraction. A calculated monosaccharide yield suggests that polymers in the HMW DOM fraction were scavenged, without substantial degradation, into the SML. Bubble scavenging experiments showed that newly aggregated particles could be formed abiotically by coagulation of low molecular weight nanometer-sized gels. Aerosol particles, artificially generated by bubbling experiments, were enriched in polysaccharides by factors of 22–70, relative to the source seawater. We propose that bubble scavenging of surface-active polysaccharides could be one of the possible mechanisms for the

  1. Polysaccharides of St. John's Wort Herb Stimulate NHDF Proliferation and NEHK Differentiation via Influence on Extracellular Structures and Signal Pathways.

    Science.gov (United States)

    Abakuks, S; Deters, A M

    2012-01-01

    St. John's Wort herb extracts often contain undesirable or volitional polysaccharides. As polysaccharides exhibit structure-dependent biological functions in the present study water-soluble polysaccharides were extracted from herb material, fractionated by anion exchange chromatography into four main polysaccharide fractions (denominated as Hp1, Hp2, Hp3 and Hp4) and characterized by HPAEC-PAD, CE, IR and GC-MS. Biological activity on human skin keratinocytes and fibroblasts was assessed by investigation of their effect on proliferation, metabolism, cytotoxicity, apoptosis and differentiation. The underlying mechanisms were investigated in gene expression studies. Polysaccharide fraction Hp1 was mainly composed of β-D-glucose. Hp2, Hp3 and Hp4 contained pectic structures and arabinogalactan proteins varying in composition and quantity. Polysaccharides of Hp1 induced the keratinocyte differentiation by inhibiting the gene expression of the epidermal growth factor and insulin receptor. While the collagen secretion of fibroblasts was stimulated by each polysaccharide fraction only Hp1 stimulated the synthesis. The fibroblast proliferation was reduced by Hp1 and increased by Hp4. This effect was related to the influence on genes that referred to oxidative stress, metabolism, transcription processes and extracellular proteins. In conclusion polysaccharides have been shown as biologically active ingredients of aqueous St. John's Wort extracts with a relation between their structural characteristics and function.

  2. Highly virulent strains of Pseudomonas solanacearum that are defective in extracellular-polysaccharide production

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Peilin; Iwata, Michiaki; Sequeira, L. (Dept. of Agriculture (USA)); Leong, S. (Univ. of Wisconsin, Madison (USA))

    1990-07-01

    Extracellular polysaccharide (EPS) has long been regarded as one of the mos important factors involved in wilting of plants by Pseudomonas solanacearum. By means of transposon Tn5 mutagensis, the authors have isolated a class of mutants that have an afluidal colony morphology but retain the ability to cause severe wilting and death of tobacco plants. One such mutant, KD700, was studied in detail. By marker exchange mutagenesis, the altered colony morphology was shown to be the result of a single Tn5 insertion in a 14.3-kilobase EcoRI fragment. This defect could be corrected by introducing a homologous clone from a cosmid library of the wild-type, parental strain K60. The Tn5-containing fragment was introduced into other P. solanacearum wild-type strains by marker exchange, and these altered strains had the same afluidal phenotype as KD700. N-Acetylgalactosamine (GalNac), the major constituent of EPS of all wild-type strains of P. solanacearum, was not detected by gas chromatography-mass spectrometry analysis of vascular fluids from wilting plants infected by KD700. In contrast, GalNac was readily detected in similar fluids of plants infected by K60. Polysaccharides extracted from culture filtrates of KD700 contained approximately one-fifth of the GalNac present in polysaccharides from K60. No differences in growth rates in culture or in planta between the mutant and the parental strains were observed. Since strains that are deficient in EPS production can remain highly virulent to tobacco, they conclude that EPS, or at least its GalNac-containing component, may not be required for disease development by P. solanacearum.

  3. Cyclic-di-GMP signalling meets extracellular polysaccharide synthesis in Bacillus subtilis.

    Science.gov (United States)

    Kampf, Jan; Stülke, Jörg

    2017-06-01

    In order to resist harmful environmental conditions, many bacteria form multicellular aggregates called biofilms. In these biofilms, they protect themselves in a self-produced matrix consisting of extracellular polysaccharides, proteins and DNA. In many bacteria, biofilm formation is stimulated in the presence of the second messenger cyclic di-GMP. In this issue of Environmental Microbiology Reports, Bedrunka and Graumann have studied matrix production by the proteins encoded in the Bacillus subtilis ydaJKLMN operon. For the first time, they were able to provide a link between c-di-GMP signalling and matrix production in this bacterium. The work demonstrates that the c-di-GMP receptor protein YdaK forms a membrane-bound complex with the YdaM and YdaN proteins, and that this interaction with YdaK is required for polysaccharide production by YdaL, YdaM and YdaN. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  4. Regulation of oxidative response and extracellular polysaccharide synthesis by a diadenylate cyclase in Streptococcus mutans.

    Science.gov (United States)

    Cheng, Xingqun; Zheng, Xin; Zhou, Xuedong; Zeng, Jumei; Ren, Zhi; Xu, Xin; Cheng, Lei; Li, Mingyun; Li, Jiyao; Li, Yuqing

    2016-03-01

    Cyclic diadenosine monophosphate (c-di-AMP) has been implicated in the control of many important bacterial activities. However, the function of this molecule in Streptococcus mutans, the primary aetiological agent of human dental caries, is unknown. In this study, we identified and characterized a diadenylate cyclase, named CdaA, in S. mutans. Furthermore, we showed that in-frame deletion of the cdaA gene in S. mutans causes decreased c-di-AMP levels, increased sensitivity to hydrogen peroxide and increased production of extracellular polysaccharides. Global gene expression profiling revealed that more than 200 genes were significantly upregulated or downregulated (> 2.0-fold) in the cdaA mutant. Interestingly, genes with increased or decreased expression were clustered in cellular polysaccharide biosynthetic processes and oxidoreductase activity respectively. Notably, the expression of several genomic islands, such as GTF-B/C, TnSmu, CRISPR1-Cas and CRISPR2-Cas, was found to be altered in the cdaA mutant, indicating a possible link between these genomic islands and c-di-AMP signalling. Collectively, the results reported here show that CdaA is an important global modulator in S. mutans and is required for optimal growth and environmental adaption. This report also paves the way to unveil further the roles of c-di-AMP signalling networks in the biology and pathogenicity of S. mutans.

  5. An electrodynamics-based model for ion diffusion in microbial polysaccharides.

    Science.gov (United States)

    Liu, Chongxuan; Zachara, John M; Felmy, Andrew; Gorby, Yuri

    2004-10-10

    An electrodynamics-based model was formulated for simulation of ion diffusion in microbial polysaccharides. The fixed charges and electrostatic double layers that may associate with microbial polysaccharides and their effects on ion diffusion were explicitly built into the model. The model extends a common multicomponent ion diffusion formulation that is based on irreversible thermodynamics under a zero ionic charge flux condition, which is only applicable to the regions without fixed charges and electrostatic double layers. An efficient numerical procedure was presented to solve the differential equations in the model. The model well described key features of experimental observations of ion diffusion in negatively charged microbial polysaccharides including accelerated diffusive transport of cations, exclusion of anions, and increased rate of cation transport with increasing negative charge density. The simulated diffusive fluxes of cations and anions were consistent with a cation exchange diffusion concept in negatively charged polysaccharides at the interface of plant roots and soils; and the developed model allows to mathematically study such diffusion phenomena. An illustrative example was also provided to simulate dynamic behavior of ionic current during ion diffusion within a charged bacterial cell wall polysaccharide and the effects of the ionic current on the compression or expansion of the bacterial electrostatic double layer at the interface of the cell wall and bulk solution.

  6. Sequestration of Reactive Blue 4 by free and immobilized Bacillus subtilis cells and its extracellular polysaccharides.

    Science.gov (United States)

    Binupriya, Arthur Raj; Sathishkumar, Muthuswamy; Ku, Chang Sub; Yun, Soon-Il

    2010-03-01

    Bacillus subtilis a gram positive bacteria and its extracellular polysaccharide were used in free form as well as immobilized form as biosorbent for sequestration of an anionic dye, Reactive Blue 4 (RB) in aqueous phase. The dye uptake enhanced with decrease in pH. Extracellular polymeric substances (EPS) and free cells were found to be better adsorbents when compared to alginate immobilized cells (IC) and EPS (IEPS). The presence of functional groups in free cells and EPS was confirmed by FT-IR analysis. Immobilization resulted in poor adsorption performance due to increase in mass transfer resistance by the polymeric matrix. High Q(max) and b values were noted in the case of free cells and free EPS in contrast to IC and IEPS. From the kinetic experiments, the adsorption system was found to be a pseudo-first-order reaction at low dye concentration. Desorption of RB was found to be 100% in 1N NaOH. However, the alginate beads were found to be unstable under high alkaline conditions of NaOH.

  7. Effects of sand burial on biomass, chlorophyll fluorescence and extracellular polysaccharides of man-made cyanobacterial crusts under experimental conditions

    Institute of Scientific and Technical Information of China (English)

    WANG WeiBo; YANG CuiYun; TANG DongShan; LI DunHai; LIU YongDing; HU ChunXiang

    2007-01-01

    Soil cyanobacterial crusts occur throughout the world, especially in the semiarid and arid regions. It always encounters sand burial, which is an important feature of mobile sand dunes. A greenhouse study was conducted to determine the effects of sand burial on biomass, chlorophyll fluorescence andextracellular polysaccharides of man-made cyanobacterial crusts in six periods of time (0, 5, 10, 15, 20 and 30 d after burying) and at five depths (0, 0.2, 0.5, 1 and 2cm). The results indicated that with the increase of the burial time and burial depth extracellular polysaccharides content and Fv/Fm decreased correspondingly and there were no significant differences between 20 and 30 burial days under different burial depths. The degradation of chlorophyll a content appeared only at 20 and 30 burial days and there was also no significant difference between them under different burial depths. It was also observed a simultaneous decrease of the values of the Fv/Fm and the content of extracellular polysaccharides happened in the crusted cyanobacterium Microcoleus vaginatus Gom. It may suggest that there exists a relationship between extracellular polysaccharides and recovery of the activity of photosystem Ⅱ (PS Ⅱ) after rehydration.

  8. Comparison of two chemical extraction methods for proteins and polysaccharides of Spirogyra fluviatilis in extracellular polymeric substances

    Science.gov (United States)

    Chang, Shuiping; Lee, Yichao

    2017-05-01

    Under natural circumstances, Spirogyra fluviatilis excretes large amounts of extracellular polymeric substances (EPS) and forms a mucilage layer. This study used two algal EPS extraction methods such as NH4OH and formaldehyde-NaOH to compare the extraction yields of EPS and the levels of its primary components of polysaccharides and proteins. The results indicated that the extraction yields of EPS using formaldehyde-NaOH and NH4OH are 8.05 mg EPS/g-fresh algae and 5.26 mg EPS/g-fresh algae, respectively. The ratio of proteins to polysaccharides in EPS is around 0.24 to 0.34, which is not a significant difference between the two extraction methods. In other words, the level of polysaccharides is always higher than the level of proteins in EPS extraction with both two methods

  9. Extracellular polysaccharides produced by Ganoderma formosanum stimulate macrophage activation via multiple pattern-recognition receptors

    Directory of Open Access Journals (Sweden)

    Wang Cheng-Li

    2012-08-01

    Full Text Available Abstract Background The fungus of Ganoderma is a traditional medicine in Asia with a variety of pharmacological functions including anti-cancer activities. We have purified an extracellular heteropolysaccharide fraction, PS-F2, from the submerged mycelia culture of G. formosanum and shown that PS-F2 exhibits immunostimulatory activities. In this study, we investigated the molecular mechanisms of immunostimulation by PS-F2. Results PS-F2-stimulated TNF-α production in macrophages was significantly reduced in the presence of blocking antibodies for Dectin-1 and complement receptor 3 (CR3, laminarin, or piceatannol (a spleen tyrosine kinase inhibitor, suggesting that PS-F2 recognition by macrophages is mediated by Dectin-1 and CR3 receptors. In addition, the stimulatory effect of PS-F2 was attenuated in the bone marrow-derived macrophages from C3H/HeJ mice which lack functional Toll-like receptor 4 (TLR4. PS-F2 stimulation triggered the phosphorylation of mitogen-activated protein kinases JNK, p38, and ERK, as well as the nuclear translocation of NF-κB, which all played essential roles in activating TNF-α expression. Conclusions Our results indicate that the extracellular polysaccharides produced by G. formosanum stimulate macrophages via the engagement of multiple pattern-recognition receptors including Dectin-1, CR3 and TLR4, resulting in the activation of Syk, JNK, p38, ERK, and NK-κB and the production of TNF-α.

  10. Nicotine promotes Streptococcus mutans extracellular polysaccharide synthesis, cell aggregation and overall lactate dehydrogenase activity.

    Science.gov (United States)

    Huang, R; Li, M; Gregory, R L

    2015-08-01

    Several epidemiology studies have reported a positive relationship between smoking and dental caries. Nicotine, an alkaloid component of tobacco, has been demonstrated to stimulate biofilm formation and metabolic activity of Streptococcus mutans, one of the most important pathogens of dental caries. The first aim of the present study was to explore the possible mechanisms leading to increased biofilm by nicotine treatment from three aspects, extracellular polysaccharides (EPS) synthesis, glucosyltransferase (Gtf) synthesis and glucan-binding protein (Gbp) synthesis at the mRNA and protein levels. The second aim was to investigate how nicotine affects S. mutans virulence, particular in lactate dehydrogenase (LDH) activity. Confocal laser scanning microscopy results demonstrated that both biofilm bacterial cell numbers and EPS were increased by nicotine. Gtf and GbpA protein expression of S. mutans planktonic cells were upregulated while GbpB protein expression of biofilm cells were downregulated by nicotine. The mRNA expression trends of those genes were mostly consistent with results on protein level but not statistically significant, and gtfD and gbpD of biofilm cells were inhibited. Nicotine was not directly involved in S. mutans LDH activity. However, since it increases the total number of bacterial cells in biofilm, the overall LDH activity of S. mutans biofilm is increased. In conclusion, nicotine stimulates S. mutans planktonic cell Gtf and Gbp expression. This leads to more planktonic cells attaching to the dental biofilm. Increased cell numbers within biofilm results in higher overall LDH activity. This contributes to caries development in smokers.

  11. Production of extracellular polysaccharide by Bacillus megaterium RB-05 using jute as substrate.

    Science.gov (United States)

    Chowdhury, Sougata Roy; Basak, Ratan Kumar; Sen, Ramkrishna; Adhikari, Basudam

    2011-06-01

    Bacillus megaterium RB-05 was grown on glucose and on "tossa-daisee" (Corchorus olitorius)-derived jute, and production and composition of extracellular polysaccharide (EPS) were monitored. An EPS yield of 0.065 ± 0.013 and of 0.297 g ± 0.054 g(-1) substrate after 72 h was obtained for glucose and jute, respectively. EPS production in the presence of jute paralleled bacterial cellulase activity. High performance liquid chromatography (HPLC), matrix assisted LASER desorption/ionization-time of flight (MALDI-ToF) mass spectroscopy, and fourier transform infrared (FT-IR) spectroscopy demonstrated that the EPS synthesized in jute culture (JC) differed from that synthesized in glucose mineral salts medium (GMSM). While fucose was only a minor constituent (4.9 wt.%) of EPS from GMSM, it a major component (41.9 wt.%) of EPS synthesized in JC. This study establishes jute as an effective fermentation substrate for EPS production by a cellulase-producing bacterium. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. Crucial Role of Extracellular Polysaccharides in Desiccation and Freezing Tolerance in the Terrestrial Cyanobacterium Nostoc commune

    Science.gov (United States)

    Tamaru, Yoshiyuki; Takani, Yayoi; Yoshida, Takayuki; Sakamoto, Toshio

    2005-01-01

    The cyanobacterium Nostoc commune is adapted to the terrestrial environment and has a cosmopolitan distribution. In this study, the role of extracellular polysaccharides (EPS) in the desiccation tolerance of photosynthesis in N. commune was examined. Although photosynthetic O2 evolution was not detected in desiccated colonies, the ability of the cells to evolve O2 rapidly recovered after rehydration. The air-dried colonies contained approximately 10% (wt/wt) water, and field-isolated, natural colonies with EPS were highly water absorbent and were rapidly hydrated by atmospheric moisture. The cells embedded in EPS in Nostoc colonies were highly desiccation tolerant, and O2 evolution was not damaged by air drying. Although N. commune was determined to be a mesophilic cyanobacterium, the cells with EPS were heat tolerant in a desiccated state. EPS could be removed from cells by homogenizing colonies with a blender and filtering with coarse filter paper. This treatment to remove EPS did not damage Nostoc cells or their ability to evolve O2, but O2 evolution was significantly damaged by desiccation treatment of the EPS-depleted cells. Similar to the EPS-depleted cells, the laboratory culture strain KU002 had only small amount of EPS and was highly sensitive to desiccation. In the EPS-depleted cells, O2 evolution was also sensitive to freeze-thaw treatment. These results strongly suggest that EPS of N. commune is crucial for the stress tolerance of photosynthesis during desiccation and during freezing and thawing. PMID:16269775

  13. Effects of lead(II) on the extracellular polysaccharide (EPS) production and colony formation of cultured Microcystis aeruginosa.

    Science.gov (United States)

    Bi, Xiang-dong; Zhang, Shu-lin; Dai, Wei; Xing, Ke-zhing; Yang, Fan

    2013-01-01

    To investigate the effects of lead(II) on the production of extracellular polysaccharides (EPS), including bound extracellular polysaccharides (bEPS) and soluble extracellular polysaccharides (sEPS), and the colony formation of Microcystis aeruginosa, cultures of M. aeruginosa were exposed to four concentrations (5.0, 10.0, 20.0 and 40.0 mg/L) of lead(II) for 10 d under controlled laboratory conditions. The results showed that 5.0 and 10.0 mg/L lead(II) stimulated M. aeruginosa growth throughout the experiment while 20.0 and 40.0 mg/L lead(II) inhibited M. aeruginosa growth in the first 2 d exposure and then stimulated it. As compared to the control group, significant increases in the bEPS and sEPS production were observed in 20.0 and 40.0 mg/L lead(II) treatments (P bEPS production, which conversely promoted colony formation, suggesting that heavy metals might be contributing to the bloom-forming of M. aeruginosa in natural conditions.

  14. The characteristics of extracellular polymeric substances and soluble microbial products in moving bed biofilm reactor-membrane bioreactor.

    Science.gov (United States)

    Duan, Liang; Jiang, Wei; Song, Yonghui; Xia, Siqing; Hermanowicz, Slawomir W

    2013-11-01

    The characteristics of extracellular polymeric substances (EPS) and soluble microbial products (SMP) in conventional membrane bioreactor (MBR) and in moving bed biofilm reactor-membrane bioreactors (MBBR-MBR) were investigated in long-term (170 days) experiments. The results showed that all reactors had high removal efficiency of ammonium and COD, despite very different fouling conditions. The MBBR-MBR with media fill ratio of 26.7% had much lower total membrane resistance and no obvious fouling were detected during the whole operation. In contrast, MBR and MBBR-MBR with lower and higher media fill experienced more significant fouling. Low fouling at optimum fill ratio may be due to the higher percentage of small molecular size (100 kDa) of EPS and SMP in the reactor. The composition of EPS and SMP affected fouling due to different O-H bonds in hydroxyl functional groups, and less polysaccharides and lipids.

  15. Characterization of Co-Cultivation of Cyanobacteria on Growth, Productions of Polysaccharides and Extracellular Proteins, Nitrogenase Activity, and Photosynthetic Activity.

    Science.gov (United States)

    Xue, Chuizhao; Wang, Libo; Wu, Tong; Zhang, Shiping; Tang, Tao; Wang, Liang; Zhao, Quanyu; Sun, Yuhan

    2017-01-01

    Cyanobacteria as biofertilizers are benefit to reduce the use of chemical fertilizers and reestablish the ecological system in soil. In general, several strains of cyanobacteria were involved in the biofertilizers. The co-cultivation of cyanobacteria were characterized on growth profile, production of polysaccharides and extracellular proteins, nitrogenase activity, and photosynthetic activity for three selected N2-fixing cyanobacteria, Anabaena cylindrica (B1611 and F243) and Nostoc sp. (F280). After eight-day culture, the highest dry weights were obtained in F280 pure culture and co-cultivation of B1611 and F280. Higher production of extracellular proteins and cell-bonding polysaccharides (CPS) were observed in co-cultivations compared with pure culture. The highest released polysaccharides (RPS) contents were obtained in pure culture of F280 and co-cultivation of F280 and F243. Galactose and glucose were major components of CPS and RPS in all samples. Trehalose was a specific component of RPS in F280 pure culture. Based on the monosaccharide contents of CPS and RPS, F280 was the dominant species in the related treatments of co-cultivation. The nitrogenase activities in all treatments exhibited a sharp rise at the late stage while a significant decrease existed when three cyanobacteria strains were mixed. Photosynthetic activities for all treatments were determined with rapid light curve, and the related parameters were estimated.

  16. Molecular and chemical features of the excreted extracellular polysaccharides in Induced Biological Soil Crusts of different ages

    Science.gov (United States)

    Rossi, Federico; Lanzhou, Chen; Liu, Yongding; Adessi, Alessandra; De Philippis, Roberto

    2014-05-01

    , assessing the activity of two key enzymes for sugar degradation: dehydrogenase and sucrase. The results obtained demonstrated a high complexity in terms of monosaccharidic composition and molecular weight, the latter resulting differently distributed between the two fractions. Enzymatic activity resulted mainly directed to the more soluble, low - molecular weight carbohydrates. The data presented represent a first study of the biochemical processes involving carbon from EPS released by IBSCs on bare substrates after the colonization of soils by the inoculated cyanobacteria. Bowker MA (2007) Biological soil crusts rehabilitation in theory and practice: an underexploited opportunity. Restoration Ecology 15(1): 13 - 23. Mager DM, Thomas AD (2011) Extracellular polysaccharides from cyanobacterial soil crusts: A review of their role in dryland soil processes. Journal of Arid Environments 75: 91 - 97. Rossi F, Diels L, Olguin E, De Philippis R (2013) Microbial fixation of CO2 in water bodies and in drylands to combat climate change, soil loss and desertification. New Biotechnology. DOI: . http://dx.doi.org/10.1016/j.nbt.2013.12.002.

  17. Microbial Decomposition of Extracellular DNA in Clay Soils

    Science.gov (United States)

    Morrissey, E. M.; McHugh, T. A.; Schwartz, E.; Preteska, L.; Hayer, M.; Hungate, B. A.

    2014-12-01

    Genomic analysis of soil communities can only be useful in predicting ecosystem processes if the genetic data gathered is representative of the microbial community. Consequently, extracellular DNA (eDNA) represents a pool of unexpressed genetic information that may skew genomic analyses. To date, our understanding of the representation of eDNA in metagenomic data and its decomposition in soil is very limited. To address this deficit, we performed a laboratory experiment wherein soils were amended with eDNA and/or clay minerals in a full factorial design. Specifically, the decomposition of 13C labeled E. coli DNA was monitored over a 30-day period in control, Kaolinite-amended, and Montmorillonite-amended soils. The amount of added eDNA carbon (C) remaining in the soil declined exponentially over time, with the majority of decomposition occurring in the first two weeks. Kaolinite significantly decreased eDNA decomposition rates and retained a higher fraction of eDNA-C (~70% remaining) than unamended and Montmorillonite-soils (~40% remaining) after 30 days. Phylogenetic (16S rRNA) sequencing of DNA extracted over the course of the incubation period enabled detection of the added eDNA. The relative abundance of added E. coli DNA decreased ~10-100 fold over 30 days. These results indicate that while a significant fraction of eDNA-C remained in the soil, this carbon was likely no longer in the form of intact strands of DNA amenable to sequencing. In addition, the eDNA affected the composition of the bacterial community. Specifically, the relative abundance of Planctomycetes and TM7 were elevated in soils that received eDNA regardless of clay addition, suggesting these phyla may be particularly effective at degrading eDNA and using it for growth. In conclusion these results indicate that the representation of eDNA in metagenomic sequence data declines rapidly, likely due to fragmentation. However, a fraction of eDNA material was resistant to decomposition, suggesting a

  18. Photorhabdus adhesion modification protein (Pam) binds extracellular polysaccharide and alters bacterial attachment

    LENUS (Irish Health Repository)

    Jones, Robert T

    2010-05-12

    Abstract Background Photorhabdus are Gram-negative nematode-symbiotic and insect-pathogenic bacteria. The species Photorhabdus asymbiotica is able to infect humans as well as insects. We investigated the secreted proteome of a clinical isolate of P. asymbiotica at different temperatures in order to identify proteins relevant to the infection of the two different hosts. Results A comparison of the proteins secreted by a clinical isolate of P. asymbiotica at simulated insect (28°C) and human (37°C) temperatures led to the identification of a small and highly abundant protein, designated Pam, that is only secreted at the lower temperature. The pam gene is present in all Photorhabdus strains tested and shows a high level of conservation across the whole genus, suggesting it is both ancestral to the genus and probably important to the biology of the bacterium. The Pam protein shows limited sequence similarity to the 13.6 kDa component of a binary toxin of Bacillus thuringiensis. Nevertheless, injection or feeding of heterologously produced Pam showed no insecticidal activity to either Galleria mellonella or Manduca sexta larvae. In bacterial colonies, Pam is associated with an extracellular polysaccharide (EPS)-like matrix, and modifies the ability of wild-type cells to attach to an artificial surface. Interestingly, Surface Plasmon Resonance (SPR) binding studies revealed that the Pam protein itself has adhesive properties. Although Pam is produced throughout insect infection, genetic knockout does not affect either insect virulence or the ability of P. luminescens to form a symbiotic association with its host nematode, Heterorhabditis bacteriophora. Conclusions We studied a highly abundant protein, Pam, which is secreted in a temperature-dependent manner in P. asymbiotica. Our findings indicate that Pam plays an important role in enhancing surface attachment in insect blood. Its association with exopolysaccharide suggests it may exert its effect through mediation of

  19. Photorhabdus adhesion modification protein (Pam binds extracellular polysaccharide and alters bacterial attachment

    Directory of Open Access Journals (Sweden)

    Joyce Susan A

    2010-05-01

    Full Text Available Abstract Background Photorhabdus are Gram-negative nematode-symbiotic and insect-pathogenic bacteria. The species Photorhabdus asymbiotica is able to infect humans as well as insects. We investigated the secreted proteome of a clinical isolate of P. asymbiotica at different temperatures in order to identify proteins relevant to the infection of the two different hosts. Results A comparison of the proteins secreted by a clinical isolate of P. asymbiotica at simulated insect (28°C and human (37°C temperatures led to the identification of a small and highly abundant protein, designated Pam, that is only secreted at the lower temperature. The pam gene is present in all Photorhabdus strains tested and shows a high level of conservation across the whole genus, suggesting it is both ancestral to the genus and probably important to the biology of the bacterium. The Pam protein shows limited sequence similarity to the 13.6 kDa component of a binary toxin of Bacillus thuringiensis. Nevertheless, injection or feeding of heterologously produced Pam showed no insecticidal activity to either Galleria mellonella or Manduca sexta larvae. In bacterial colonies, Pam is associated with an extracellular polysaccharide (EPS-like matrix, and modifies the ability of wild-type cells to attach to an artificial surface. Interestingly, Surface Plasmon Resonance (SPR binding studies revealed that the Pam protein itself has adhesive properties. Although Pam is produced throughout insect infection, genetic knockout does not affect either insect virulence or the ability of P. luminescens to form a symbiotic association with its host nematode, Heterorhabditis bacteriophora. Conclusions We studied a highly abundant protein, Pam, which is secreted in a temperature-dependent manner in P. asymbiotica. Our findings indicate that Pam plays an important role in enhancing surface attachment in insect blood. Its association with exopolysaccharide suggests it may exert its effect

  20. Effects of light intensity on components and topographical structures of extracellular polysaccharides from the cyanobacteria Nostoc sp.

    Science.gov (United States)

    Ge, Hongmei; Xia, Ling; Zhou, Xuping; Zhang, Delu; Hu, Chunxiang

    2014-02-01

    A study on the effects of light intensity (40 and 80 μE/m(2)/sec) on the components and topographical structures of extracellular polysaccharides (EPS) was carried out in cyanobacteria Nostoc sp.. EPS yield increased with light intensity. However, light intensity did not significantly affect the EPS fractions and monosaccharide composition. Higher light intensity generally resulted in higher protein content of EPS in similar fractions. The topographical structure of EPS, investigated by atomic force microscopy, appeared as spherical lumps, chains and networks. The long chains were observed at higher light intensity. Thus, light intensity affected the yield and nature of EPS.

  1. Extracellular polysaccharides do not inhibit the reaction between Streptococcus mutans and its specific immunoglobulin G (IgG) or penetration of the IgG through S. mutans biofilm.

    Science.gov (United States)

    Zhu, M; Takenaka, S; Sato, M; Hoshino, E

    2001-02-01

    The present study investigated whether extracellular polysaccharides inhibit reaction between Streptococcus mutans and its specific immunoglobulin G (IgG) and penetration of the IgG through S. mutans biofilm. The planktonic organisms with or without extracellular polysaccharides were prepared, incubated with rabbit IgG against whole cell of S. mutans and fluorescein isothiocyanate (FITC)-conjugated goat affinity purified antibody to rabbit IgG. Biofilms with or without extracellular polysaccharides were formed on cover glasses and incubated with rabbit IgG against S. mutans and FITC-conjugated goat antibody to rabbit IgG. Then, biofilms were stained with propidium iodide. The amount of specific IgG binding on S. mutans was determined by FITC intensity with a fluorescence microplate reader. The penetration of IgG through biofilms was determined by confocal laser scanning microscopy. The results showed that the fluorescence intensity of FITC in planktonic organisms with extracellular polysaccharides was similar to that in planktonic organisms without extracellular polysaccharides, indicating that extracellular polysaccharides did not inhibit the reaction between S. mutans and its specific IgG. Although biofilms of S. mutans with extracellular polysaccharides were much thicker and denser than those without extracellular polysaccharides, the speed with which IgG penetrated through both of the biofilms did not differ significantly, suggesting that penetration of IgG through S. mutans biofilm was not affected by extracellular polysaccharides.

  2. Extracellular Enzyme Activity assay as indicator of soil microbial functional diversity and activity

    DEFF Research Database (Denmark)

    Hendriksen, Niels Bohse; Winding, Anne

    2012-01-01

    and especially hydrolytic extracellular enzymes are of pivotal importance for decomposition of organic substrates and biogeochemical cycling. Their activity reflects the functional diversity and activity of the microorganisms involved in decomposition processes which are essential processes for soil functioning...... and soil ecosystem services. The soil enzyme activity has been measured by the use of fluorogenic model substrates e.g. methylumbelliferyl (MUF) substrates for a number of enzymes involved in the degradation of polysaccharides as cellulose, hemicellulose and chitin, while degradation of proteins has been...... followed by amino-methyl-coumaric substrates (AMC). The already developed Extracellular Enzyme Activity (EEA) assay was further optimized as a microwell based assay for the activity of enzymes involved in degradation of polysaccharides and proteins. Using specific MUF and AMC substrates on European soils...

  3. Withania somnifera attenuates acid production, acid tolerance and extra-cellular polysaccharide formation of Streptococcus mutans biofilms.

    Science.gov (United States)

    Pandit, Santosh; Song, Kwang-Yeob; Jeon, Jae-Gyu

    2014-01-01

    Withania somnifera (Ashwagandha) is a plant of the Solanaceae family. It has been widely used as a remedy for a variety of ailments in India and Nepal. The plant has also been used as a controlling agent for dental diseases. The aim of the present study was to evaluate the activity of the methanol extract of W. somnifera against the physiological ability of cariogenic biofilms and to identify the components of the extract. To determine the activity of the extract, assays for sucrose-dependent bacterial adherence, glycolytic acid production, acid tolerance, and extracellular polysaccharide formation were performed using Streptococcus mutans biofilms. The viability change of S. mutans biofilms cells was also determined. A phytochemical analysis of the extract was performed using TLC and LC/MS/MS. The extract showed inhibitory effects on sucrose-dependent bacterial adherence (≥ 100 μg/ml), glycolytic acid production (≥ 300 μg/ml), acid tolerance (≥ 300 μg/ml), and extracellular polysaccharide formation (≥ 300 μg/ml) of S. mutans biofilms. However, the extract did not alter the viability of S. mutans biofilms cells in all concentrations tested. Based on the phytochemical analysis, the activity of the extract may be related to the presence of alkaloids, anthrones, coumarines, anthraquinones, terpenoids, flavonoids, and steroid lactones (withanolide A, withaferin A, withanolide B, withanoside IV, and 12-deoxy withastramonolide). These data indicate that W. somnifera may be a potential agent for restraining the physiological ability of cariogenic biofilms.

  4. Structural characterization and anti-aging activity of a novel extracellular polysaccharide from fungus Phellinus sp. in a mammalian system.

    Science.gov (United States)

    Ma, Xiao-Kui; Guo, Dan Dan; Peterson, Eric Charles; Dun, Ying; Li, Dan Yang

    2016-08-10

    Little is known about the chemical structure of purified extracellular polysaccharides from Phellinus sp., a fungal species with known medicinal properties. A combination of IR spectroscopy, methylation analysis and NMR were performed for the structural analysis of a purified extracellular polysaccharide derived from Phellinus sp. culture, denoted as SHP-1, along with an evaluation of the anti-aging effect in vivo of the polysaccharide supplementation. The structure of SHP-1 was established, with a backbone composed of →2,4)-α-d-glucopyranose-(1→ and →2)-β-d-mannopyranose-(1→ and two terminal glucopyranose branches. Biochemical analysis from mammalian animal experiments demonstrated that SHP-1 possesses the ability to enhance antioxidant enzyme activities, such as catalase (CAT) and superoxide dismutase (SOD) activities, Trolox equivalent antioxidant capacity (TEAC) in serum of d-galactose-aged mice, while reducing lipofuscin levels, another indicator of cell aging, indicating a potential association with anti-aging activities in a dose dependent manner. This compound had a favourable influence on immune organ indices, and a marked amelioration ability of histopathological hepatic lesions such as necrosis, karyolysis and reduced inflammation and apoptosis in mouse hepatocytes. These results suggest that SHP-1 has strong antioxidant activities and a significant protective effect against oxidative stress or hepatotoxicity induced by d-galactose in mice and it could be developed as a food ingredient or a pharmaceutical to prevent many age-associated diseases such as major depressive disorder and hepatotoxicity. To our knowledge, this is the first report on the antioxidant effects of a novel purified exopolysaccharide derived from Phellinus sp.

  5. Polysaccharides of St. John’s Wort Herb Stimulate NHDF Proliferation and NEHK Differentiation via Influence on Extracellular Structures and Signal Pathways

    Directory of Open Access Journals (Sweden)

    S. Abakuks

    2012-01-01

    Full Text Available St. John's Wort herb extracts often contain undesirable or volitional polysaccharides. As polysaccharides exhibit structure-dependent biological functions in the present study water-soluble polysaccharides were extracted from herb material, fractionated by anion exchange chromatography into four main polysaccharide fractions (denominated as Hp1, Hp2, Hp3 and Hp4 and characterized by HPAEC-PAD, CE, IR and GC-MS. Biological activity on human skin keratinocytes and fibroblasts was assessed by investigation of their effect on proliferation, metabolism, cytotoxicity, apoptosis and differentiation. The underlying mechanisms were investigated in gene expression studies. Polysaccharide fraction Hp1 was mainly composed of β-D-glucose. Hp2, Hp3 and Hp4 contained pectic structures and arabinogalactan proteins varying in composition and quantity. Polysaccharides of Hp1 induced the keratinocyte differentiation by inhibiting the gene expression of the epidermal growth factor and insulin receptor. While the collagen secretion of fibroblasts was stimulated by each polysaccharide fraction only Hp1 stimulated the synthesis. The fibroblast proliferation was reduced by Hp1 and increased by Hp4. This effect was related to the influence on genes that referred to oxidative stress, metabolism, transcription processes and extracellular proteins. In conclusion polysaccharides have been shown as biologically active ingredients of aqueous St. John's Wort extracts with a relation between their structural characteristics and function.

  6. Microbial respiration and extracellular enzyme activity in sediments from the Gulf of Mexico hypoxic zone

    Science.gov (United States)

    This study explores the relationship between sediment chemistry (TC, TN, TP) and microbial respiration (DHA) and extracellular enzyme activity (EEA) across the Gulf of Mexico (GOM) hypoxic zone. TC, TN, and TP were all positively correlated with each other (r=0.19-0.68). DHA was ...

  7. Microbial respiration and extracellular enzyme activity in sediments from the Gulf of Mexico hypoxic zone

    Science.gov (United States)

    This study explores the relationship between sediment chemistry (TC, TN, TP) and microbial respiration (DHA) and extracellular enzyme activity (EEA) across the Gulf of Mexico (GOM) hypoxic zone. TC, TN, and TP were all positively correlated with each other (r=0.19-0.68). DHA was ...

  8. Abiotic factors in colony formation: effects of nutrition and light on extracellular polysaccharide production and cell aggregates of Microcystis aeruginosa

    Science.gov (United States)

    Yang, Zhen; Kong, Fanxiang

    2013-07-01

    Colony morphology is important for Microcystis to sustain a competitive advantage in eutrophic lakes. The mechanism of colony formation in Microcystis is currently unclear. Extracellular polysaccharide (EPS) has been reported to play an important role in cell aggregate formation of some phytoplankton. Microcystis aeruginosa was cultivated under varied abiotic conditions, including different nutrient, light, and temperature conditions, to investigate their effects on EPS production and morphological change. The results show that nutrient concentration and light intensity have great effects on EPS productionin M. aeruginosa. There was a considerable increase in EPS production after M. aeruginosa was cultivated in adjusted culture conditions similar to those present in the field (28.9 mg C/L, 1.98 mg N/L, 0.65 mg P/L, light intensity: 100 μmol/(m2 · s)). These results indicate that abiotic factors might be one of the triggers for colony formation in Microcystis.

  9. Effects of sand burial on biomass, chlorophyll fluores-cence and extracellular polysaccharides of man-made cyanobacterial crusts under experimental conditions

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Soil cyanobacterial crusts occur throughout the world, especially in the semiarid and arid regions. It always encounters sand burial, which is an important feature of mobile sand dunes. A greenhouse study was conducted to determine the effects of sand burial on biomass, chlorophyll fluorescence and extracellular polysaccharides of man-made cyanobacterial crusts in six periods of time (0, 5, 10, 15, 20 and 30 d after burying) and at five depths (0, 0.2, 0.5, 1 and 2cm). The results indicated that with the increase of the burial time and burial depth extracellular polysaccharides content and Fv/Fm decreased correspondingly and there were no significant differences between 20 and 30 burial days under dif-ferent burial depths. The degradation of chlorophyll a content appeared only at 20 and 30 burial days and there was also no significant difference between them under different burial depths. It was also observed a simultaneous decrease of the values of the Fv/Fm and the content of extracellular poly-saccharides happened in the crusted cyanobacterium Microcoleus vaginatus Gom. It may suggest that there exists a relationship between extracellular polysaccharides and recovery of the activity of pho-tosystem II (PS II) after rehydration.

  10. The Azospirillum brasilense Sp7 noeJ and noeL genes are involved in extracellular polysaccharide biosynthesis.

    Science.gov (United States)

    Lerner, Anat; Castro-Sowinski, Susana; Valverde, Angel; Lerner, Hadas; Dror, Rachel; Okon, Yaacov; Burdman, Saul

    2009-12-01

    Azospirillum brasilense is a plant root-colonizing bacterium that exerts beneficial effects on the growth of many agricultural crops. Extracellular polysaccharides of the bacterium play an important role in its interactions with plant roots. The pRhico plasmid of A. brasilense Sp7, also named p90, carries several genes involved in synthesis and export of cell surface polysaccharides. We generated two Sp7 mutants impaired in two pRhico-located genes, noeJ and noeL, encoding mannose-6-phosphate isomerase and GDP-mannose 4,6-dehydratase, respectively. Our results demonstrate that in A. brasilense Sp7, noeJ and noeL are involved in lipopolysaccharide and exopolysaccharide synthesis. noeJ and noeL mutant strains were significantly altered in their outer membrane and cytoplasmic/periplasmic protein profiles relative to the wild-type strain. Moreover, both noeJ and noeL mutations significantly affected the bacterial responses to several stresses and antimicrobial compounds. Disruption of noeL, but not noeJ, affected the ability of the A. brasilense Sp7 to form biofilms. The pleiotropic alterations observed in the mutants could be due, at least partially, to their altered lipopolysaccharides and exopolysaccharides relative to the wild-type.

  11. Extracellular polysaccharide production by a strain of Pleurotus djamor isolated in the south of Brazil and antitumor activity on Sarcoma 180.

    Science.gov (United States)

    Borges, Gisele Martini; De Barba, Fabiana Figueredo Molin; Schiebelbein, Ana Paula; Pereira, Bruna Parmezzani; Chaves, Mariane Bonatti; Silveira, Marcia Luciane Lange; Pinho, Mauro Souza Leite; Furlan, Sandra Aparecida; Wisbeck, Elisabeth

    2013-12-01

    Polysaccharides with medicinal properties can be obtained from fruiting bodies, mycelium and culture broth of several fungus species. This work was carried out in batch culture using a stirred tank reactor with two different initial glucose concentrations (40-50 g/L) and pH values (3.0-4.0) to enhance extracellular polysaccharides production by Pleurotus djamor UNIVILLE 001 and evaluate antitumor effect of intraperitonial administration of Pleurotus djamor extract on sarcoma 180 animal model. According to factorial design, the low pH value (pH 3.0) led to a gain of 1.6 g/L on the extracellular polysaccharide concentration, while glucose concentration in the tested range had no significant effect on the concentration of polysaccharide. With 40 g/L initial glucose concentration and pH 3.0, it was observed that yield factor of extracellular polysaccharide on substrate (YP/S = 0.072) and maximum extracellular polysaccharide productivity (Q(Pmax) = 11.26 mg/L.h) were about 188% and 321% respectively higher than those obtained in the experiment performed at pH 4.0. Under these conditions, the highest values of the yield factor of biomass on substrate (YX/S = 0.24) and maximal biomass productivity (Q(Xmax) = 32.2 mg/L.h) were also reached. In tumor response study, mean tumor volume on the 21th day was 35.3 cm(3) in untreated group and 1.6 cm(3) in treated group (p = 0.05) with a tumor inhibition rate of 94%. These impressive results suggests an inhibitory effect of P.djamor extract on cancer cells.

  12. Extracellular polysaccharide production by a strain of Pleurotus djamor isolated in the south of Brazil and antitumor activity on Sarcoma 180

    Directory of Open Access Journals (Sweden)

    Gisele Martini Borges

    2013-12-01

    Full Text Available Polysaccharides with medicinal properties can be obtained from fruiting bodies, mycelium and culture broth of several fungus species. This work was carried out in batch culture using a stirred tank reactor with two different initial glucose concentrations (40-50 g/L and pH values (3.0-4.0 to enhance extracellular polysaccharides production by Pleurotus djamor UNIVILLE 001 and evaluate antitumor effect of intraperitonial administration of Pleurotus djamor extract on sarcoma 180 animal model. According to factorial design, the low pH value (pH 3.0 led to a gain of 1.6 g/L on the extracellular polysaccharide concentration, while glucose concentration in the tested range had no significant effect on the concentration of polysaccharide. With 40 g/L initial glucose concentration and pH 3.0, it was observed that yield factor of extracellular polysaccharide on substrate (Y P/S = 0.072 and maximum extracellular polysaccharide productivity (Q Pmax = 11.26 mg/L.h were about 188% and 321% respectively higher than those obtained in the experiment performed at pH 4.0. Under these conditions, the highest values of the yield factor of biomass on substrate (Y X/S = 0.24 and maximal biomass productivity (Q Xmax = 32.2 mg/L.h were also reached. In tumor response study, mean tumor volume on the 21th day was 35.3 cm³ in untreated group and 1.6 cm³ in treated group (p = 0.05 with a tumor inhibition rate of 94%. These impressive results suggests an inhibitory effect of P.djamor extract on cancer cells.

  13. Extracellular Polysaccharide Production in a Scytonemin-Deficient Mutant of Nostoc punctiforme Under UVA and Oxidative Stress.

    Science.gov (United States)

    Soule, Tanya; Shipe, Dexter; Lothamer, Justin

    2016-10-01

    Some cyanobacteria can protect themselves from ultraviolet radiation by producing sunscreen pigments. In particular, the sheath pigment scytonemin protects cells against long-wavelength UVA radiation and is only found in cyanobacteria which are capable of extracellular polysaccharide (EPS) production. The presence of a putative glycosyltransferase encoded within the scytonemin gene cluster, along with the localization of scytonemin and EPS to the extracellular sheath, prompted us to investigate the relationship between scytonemin and EPS production under UVA stress. In this study, it was hypothesized that there would be a relationship between the biosynthesis of scytonemin and EPS under both UVA and oxidative stress, since the latter is a by-product of UVA radiation. EPS production was measured following exposure of wild-type Nostoc punctiforme and the non-scytonemin-producing strain SCY59 to UVA and oxidative stress. Under UVA, SCY59 produced significantly more EPS than the unstressed controls and the wild type, while both strains produced more EPS under oxidative stress compared to the controls. The results suggest that EPS secretion occurs in response to the oxidative stress by-product of UVA rather than as a direct response to UVA radiation.

  14. Isolation and characterization of extracellular polysaccharides from the epipelic diatoms Cylindrotheca closterium and Navicula salinarum

    NARCIS (Netherlands)

    Staats, N.; de Winder, B.; Stal, L.J.; Mur, L.R.

    1999-01-01

    The production and composition of extracellular polymeric substances (EPS) in axenic batch cultures of the benthic marine epipelic diatoms Navicula salinarum and Cylindrotheca closterium were investigated. EPS was secreted into the medium and the bulk was loosely associated with the cells. Neither N

  15. Analytical methods for soluble microbial products (SMP) and extracellular polymers (ECP) in wastewater treatment systems: a review.

    Science.gov (United States)

    Kunacheva, Chinagarn; Stuckey, David C

    2014-09-15

    Effluents from biological processes contain a wide range of complex organic compounds, including soluble microbial products (SMP) and extracellular polymers (ECP), released during bacteria metabolism in mixed culture in bioreactors. It is important to clearly identify the primary components of SMPs and ECPs in order to understand the fundamental mechanisms of biological activity that create these compounds, and how to reduce these compounds in the effluent. In addition, these compounds constitute the main foulants in membrane bioreactors which are being used more widely around the world. A review on the extraction of ECP, characterization, and identification of SMPs and ECPs is presented, and we summarize up-to-date pretreatments and analytical methods for SMPs. Most researchers have focused more on the overall properties of SMPs and ECPs such as their concentrations, molecular weight distribution, aromaticity, hydrophobic and hydrophilic properties, biodegradability, and toxicity characteristics. Many studies on the identification of effluent SMPs show that most of these compounds were not present in the influent, such as humic acids, polysaccharides, proteins, nucleic acids, organic acids, amino acids, exocellular enzymes, structural components of cells and products of energy metabolism. A few groups of researchers have been working on the identification of compounds in SMPs using advanced analytical techniques such as GC-MS, LC-IT-TOF-MS and MALDI-TOF-MS. However, there is still considerably more work needed to be done analytically to fully understand the chemical characteristics of SMPs and ECPs.

  16. Characterization of an extracellular polysaccharide produced by Bacillus sp.RL-2

    Institute of Scientific and Technical Information of China (English)

    LUO Ping; LUO Gu-yuan; JI Fang-ying; CAI Jiang-wei

    2005-01-01

    A strain secreting a strongly acidic polysaccharide flocculating agent was isolated from activated sludge, and identified as Bacillus brevis. The bioflocculant was produced by RL-2 during the late logarithmic growth in the batch culture and was recovered from supernatant by ethanol precipitation. The bioflocculant is thermo-stable as its activity remains stable after heated at 100℃ for 45 min. Its flocculating activity with kaolin suspensions was stimulated by the addition of Ca2+, Al3+ and Cu2+. The flocculant consists of glucose, mannose, and galacturonic acid. Its average molecular mass was estimated to be approximately 2.86×105 by the method of viscosity. The flocculant aggregates various inorganic and organic compounds in solution.

  17. Riboflavin-shuttled extracellular electron transfer from Enterococcus faecalis to electrodes in microbial fuel cells.

    Science.gov (United States)

    Zhang, Enren; Cai, Yamin; Luo, Yue; Piao, Zhe

    2014-11-01

    Great attention has been focused on Gram-negative bacteria in the application of microbial fuel cells. In this study, the Gram-positive bacterium Enterococcus faecalis was employed in microbial fuel cells. Bacterial biofilms formed by E. faecalis ZER6 were investigated with respect to electricity production through the riboflavin-shuttled extracellular electron transfer. Trace riboflavin was shown to be essential for transferring electrons derived from the oxidation of glucose outside the peptidoglycan layer in the cell wall of E. faecalis biofilms formed on the surface of electrodes, in the absence of other potential electron mediators (e.g., yeast extract).

  18. Extracellular enzyme activity assay as indicator of soil microbial functional diversity and activity

    DEFF Research Database (Denmark)

    Hendriksen, Niels Bohse; Winding, Anne

    2012-01-01

    Extracellular enzyme activity assay as indicator of soil microbial functional diversity and activity Niels Bohse Hendriksen, Anne Winding. Department of Environmental Science, Aarhus University, 4000 Roskilde, Denmark Soils provide numerous essential ecosystem services such as carbon cycling...... the functional diversity and activity of the microorganisms involved in decomposition processes. Their activity has been measured by the use of fluorogenic model substrates e.g. methylumbelliferyl (MUF) substrates for a number of enzymes involved in the degradation of polysacharides as cellulose, hemicellulose...

  19. Structure of Plant Cell Walls : XVIII. An Analysis of the Extracellular Polysaccharides of Suspension-Cultured Sycamore Cells.

    Science.gov (United States)

    Stevenson, T T; McNeil, M; Darvill, A G; Albersheim, P

    1986-04-01

    The water-soluble polysaccharides (SEPS) secreted into the medium by suspension-cultured sycamore cells were examined to determine whether the polysaccharides were the same as those present in the walls of sycamore cells. The SEPS were made more amenable to fractionation by treatment with a highly purified alpha-1,4-endopolygalacturonase (EPG). The EPG-treated SEPS were fractionated by anion-exchange and gelpermeation chromatography. The following polysaccharides were found: xyloglucan, arabinoxylan, at least two arabinogalactans, a rhamnogalacturonan-II-like polysaccharide, and a polygalacturonic acid-rich polysaccharide. The oligogalacturonide fragments expected from EPG-digested homogalacturonan were also identified. Evidence was obtained for the presence of a rhamnogalacturonan-I-like polysaccharide. All of the above polysaccharides have been isolated from or are believed to be present in sycamore cell walls. Furthermore, all of the noncellulosic polysaccharides known to be present in sycamore cell-walls appear to be present in the SEPS.

  20. Abiotic factors in colony formation: effects of nutrition and light on extracellular polysaccharide production and cell aggregates of Microcystis aeruginosa

    Institute of Scientific and Technical Information of China (English)

    YANG Zhen; KONG Fanxiang

    2013-01-01

    Colony morphology is important for Microcystis to sustain a competitive advantage in eutrophic lakes.The mechanism of colony formation in Microcystis is currently unclear.Extracellular polysaccharide (EPS) has been reported to play an important role in cell aggregate formation of some phytoplankton.Microcystis aeruginosa was cultivated under varied abiotic conditions,including different nutrient,light,and temperature conditions,to investigate their effects on EPS production and morphological change.The results show that nutrient concentration and light intensity have great effects on EPS production in M.aeruginosa.There was a considerable increase in EPS production after M.aeruginosa was cultivated in adjusted culture conditions similar to those present in the field (28.9 mg C/L,1.98 mg N/L,0.65 mg P/L,light intensity:100 μmol/(m2·s)).These results indicate that abiotic factors might be one of the triggers for colony formation in Microcystis.

  1. Conversion of cheese whey into a fucose- and glucuronic acid-rich extracellular polysaccharide by Enterobacter A47.

    Science.gov (United States)

    Antunes, Sílvia; Freitas, Filomena; Alves, Vítor D; Grandfils, Christian; Reis, Maria A M

    2015-09-20

    Cheese whey was used as the sole substrate for the production of extracellular polysaccharides (EPS) by Enterobacter A47. An EPS concentration of 6.40 g L(-1) was reached within 3.2 days of cultivation, corresponding to a volumetric productivity of 2.00 g L(-1) d(-1). The produced EPS was mainly composed of glucuronic acid (29 mol%) and fucose (29 mol%), with lower contents of glucose and galactose (21 mol% each) and a total acyl groups content of 32 wt.%. The polymer had an average molecular weight of 1.8×10(6) Da, with a polydispersity index of 1.2, and an intrinsic viscosity of 8.0 dL g(-1). EPS aqueous solutions (1.0 wt.% in 0.01 M NaCl, at pH 8.0) presented a shear thinning behavior with a viscosity of the first Newtonian plateau approaching 0.1 Pas. This novel glucuronic acid-rich polymer possesses interesting rheological properties, which, together with its high content of glucuronic acid and fucose, two bioactive sugar monomers, confers it a great potential for use in high-value applications, such as cosmetics and pharmaceuticals.

  2. Glyceraldehyde-3-phosphate dehydrogenase of Xanthomonas campestris pv. campestris is required for extracellular polysaccharide production and full virulence.

    Science.gov (United States)

    Lu, Guang-Tao; Xie, Jia-Ri; Chen, Lei; Hu, Jiang-Ru; An, Shi-Qi; Su, Hui-Zhao; Feng, Jia-Xun; He, Yong-Qiang; Jiang, Bo-Le; Tang, Dong-Jie; Tang, Ji-Liang

    2009-05-01

    Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) plays an important role in glucose catabolism, converting glyceraldehyde 3-phosphates to 1,3-bisphosphoglycerates. Open reading frame (ORF) XC_0972 in the genome of Xanthomonas campestris pv. campestris (Xcc) strain 8004 is the only ORF in this strain annotated to encode a GAPDH. In this work, we have demonstrated genetically that this ORF encodes a unique GAPDH in Xcc strain 8004, which seems to be constitutively expressed. A GAPDH-deficient mutant could still grow in medium with glucose or other sugars as the sole carbon source, and no phosphofructokinase activity was detectable in strain 8004. These facts suggest that Xcc may employ the Entner-Doudoroff pathway, but not glycolysis, to utilize glucose. The mutant could not utilize pyruvate as sole carbon source, whereas the wild-type could, implying that the GAPDH of Xcc is involved in gluconeogenesis. Furthermore, inactivation of the Xcc GAPDH resulted in impairment of bacterial growth and virulence in the host plant, and reduction of intracellular ATP and extracellular polysaccharide (EPS). This reveals that GAPDH is required for EPS production and full pathogenicity of Xcc.

  3. N-acetyl-L-cysteine affects growth, extracellular polysaccharide production, and bacterial biofilm formation on solid surfaces.

    Science.gov (United States)

    Olofsson, Ann-Cathrin; Hermansson, Malte; Elwing, Hans

    2003-08-01

    N-Acetyl-L-cysteine (NAC) is used in medical treatment of patients with chronic bronchitis. The positive effects of NAC treatment have primarily been attributed to the mucus-dissolving properties of NAC, as well as its ability to decrease biofilm formation, which reduces bacterial infections. Our results suggest that NAC also may be an interesting candidate for use as an agent to reduce and prevent biofilm formation on stainless steel surfaces in environments typical of paper mill plants. Using 10 different bacterial strains isolated from a paper mill, we found that the mode of action of NAC is chemical, as well as biological, in the case of bacterial adhesion to stainless steel surfaces. The initial adhesion of bacteria is dependent on the wettability of the substratum. NAC was shown to bind to stainless steel, increasing the wettability of the surface. Moreover, NAC decreased bacterial adhesion and even detached bacteria that were adhering to stainless steel surfaces. Growth of various bacteria, as monocultures or in a multispecies community, was inhibited at different concentrations of NAC. We also found that there was no detectable degradation of extracellular polysaccharides (EPS) by NAC, indicating that NAC reduced the production of EPS, in most bacteria tested, even at concentrations at which growth was not affected. Altogether, the presence of NAC changes the texture of the biofilm formed and makes NAC an interesting candidate for use as a general inhibitor of formation of bacterial biofilms on stainless steel surfaces.

  4. Mutation breeding of extracellular polysaccharide-producing microalga Crypthecodinium cohnii by a novel mutagenesis with atmospheric and room temperature plasma.

    Science.gov (United States)

    Liu, Bin; Sun, Zheng; Ma, Xiaonian; Yang, Bo; Jiang, Yue; Wei, Dong; Chen, Feng

    2015-04-13

    Extracellular polysaccharides (EPS) produced by marine microalgae have the potential to be used as antioxidants, antiviral agents, immunomodulators, and anti-inflammatory agents. Although the marine microalga Crypthecodinium cohnii releases EPS during the process of docosahexaenoic acid (DHA) production, the yield of EPS remains relatively low. To improve the EPS production, a novel mutagenesis of C. cohnii was conducted by atmospheric and room temperature plasma (ARTP). Of the 12 mutants obtained, 10 mutants exhibited significantly enhanced EPS yield on biomass as compared with the wild type strain. Among them, mutant M7 was the best as it could produce an EPS volumetric yield of 1.02 g/L, EPS yield on biomass of 0.39 g/g and EPS yield on glucose of 94 mg/g, which were 33.85%, 85.35% and 57.17% higher than that of the wild type strain, respectively. Results of the present study indicated that mutagenesis of the marine microalga C. cohnii by ARTP was highly effective leading to the high-yield production of EPS.

  5. Ralstonia solanacearum extracellular polysaccharide is a specific elicitor of defense responses in wilt-resistant tomato plants.

    Directory of Open Access Journals (Sweden)

    Annett Milling

    Full Text Available Ralstonia solanacearum, which causes bacterial wilt of diverse plants, produces copious extracellular polysaccharide (EPS, a major virulence factor. The function of EPS in wilt disease is uncertain. Leading hypotheses are that EPS physically obstructs plant water transport, or that EPS cloaks the bacterium from host plant recognition and subsequent defense. Tomato plants infected with R. solanacearum race 3 biovar 2 strain UW551 and tropical strain GMI1000 upregulated genes in both the ethylene (ET and salicylic acid (SA defense signal transduction pathways. The horizontally wilt-resistant tomato line Hawaii7996 activated expression of these defense genes faster and to a greater degree in response to R. solanacearum infection than did susceptible cultivar Bonny Best. However, EPS played different roles in resistant and susceptible host responses to R. solanacearum. In susceptible plants the wild-type and eps(- mutant strains induced generally similar defense responses. But in resistant Hawaii7996 tomato plants, the wild-type pathogens induced significantly greater defense responses than the eps(- mutants, suggesting that the resistant host recognizes R. solanacearum EPS. Consistent with this idea, purified EPS triggered significant SA pathway defense gene expression in resistant, but not in susceptible, tomato plants. In addition, the eps(- mutant triggered noticeably less production of defense-associated reactive oxygen species in resistant tomato stems and leaves, despite attaining similar cell densities in planta. Collectively, these data suggest that bacterial wilt-resistant plants can specifically recognize EPS from R. solanacearum.

  6. Production and characterization of an extracellular polysaccharide of antarctic marine bacteria Pseudoalteromonas sp. S-15-13

    Institute of Scientific and Technical Information of China (English)

    LI Jiang; CHEN Kaoshan; LIN Xuezheng; HE Peiqing; LI Guangyou

    2006-01-01

    Twenty-seven antarctic bacteria producing extracellular polysaccharide (EPS) were selected from 57 strains by staining technology. The effects of major environmental factors on the growth and EPS production of Pseudoalteromonas sp. S-15-13 were investigated, and the EPS was separated and purified for characterization analysis. The results showed that the optimal conditions for the EPS production were culture period, 56 h; growth temperature, 8 ℃; carbon source, 1.0% glucose; NaCl concentration, 3.0%; pH 6.0~7.0. The EPS was purified by cold ethanol precipitation, proteins removal, ion exchange chromatography and gel chromatography technology. The molecular mass of EPS-II was 62 kDa as determined by the high performance gel permeation chromatography. Its sugar composition was a homopolymer of mannose analyzed by gas chromatograph spectroscopy. After repeated freezing and thawing of the bacteria biomass in the presence of EPS, the bacterial growth was much higher than that observed after freezing in the absence of EPS and the difference augmented with the increase of freeze-thaw cycles. It is hypothesized that the adaptation of Pseudoalteromonas sp. S-15-13 to the antarctic marine conditions, characterized by low temperature, high NaCl concentration and repeated freeze-thaw cycles, might be related to the EPS production ability.

  7. Patterns of extracellular enzyme activities and microbial metabolism in an Arctic fjord of Svalbard and in the northern Gulf of Mexico: contrasts in carbon processing by pelagic microbial communities

    Directory of Open Access Journals (Sweden)

    Carol eArnosti

    2013-10-01

    Full Text Available The microbial community composition of polar and temperate ocean waters differs substantially, but the potential functional consequences of these differences are largely unexplored. We measured bacterial production, glucose metabolism, and the abilities of microbial communities to hydrolyze a range of polysaccharides in an Arctic fjord of Svalbard (Smeerenburgfjord, and thus to initiate remineralization of high-molecular weight organic matter. We compared these data with similar measurements previously carried out in the northern Gulf of Mexico in order to investigate whether differences in the spectrum of enzyme activities measurable in Arctic and temperate environments are reflected in ‘downstream’ aspects of microbial metabolism (metabolism of monomers and biomass production. Only 4 of 6 polysaccharide substrates were hydrolyzed in Smeerenburgfjord; all were hydrolyzed in the upper water column of the Gulf. These patterns are consistent on an interannual basis. Bacterial protein production was comparable at both locations, but the pathways of glucose utilization differed. Glucose incorporation rate constants were comparatively higher in Svalbard, but glucose respiration rate constants were higher in surface waters of the Gulf. As a result, at the time of sampling ca. 75% of the glucose was incorporated into biomass in Svalbard, but in the northern Gulf of Mexico most of the glucose was respired to CO2. A limited range of enzyme activities is therefore not a sign of a dormant community or one unable to further process substrates resulting from extracellular enzymatic hydrolysis. The ultimate fate of carbohydrates in marine waters, however, is strongly dependent upon the specific capabilities of heterotrophic microbial communities in these disparate environments.

  8. Patterns of extracellular enzyme activities and microbial metabolism in an Arctic fjord of Svalbard and in the northern Gulf of Mexico: contrasts in carbon processing by pelagic microbial communities.

    Science.gov (United States)

    Arnosti, Carol; Steen, Andrew D

    2013-01-01

    The microbial community composition of polar and temperate ocean waters differs substantially, but the potential functional consequences of these differences are largely unexplored. We measured bacterial production, glucose metabolism, and the abilities of microbial communities to hydrolyze a range of polysaccharides in an Arctic fjord of Svalbard (Smeerenburg Fjord), and thus to initiate remineralization of high-molecular weight organic matter. We compared these data with similar measurements previously carried out in the northern Gulf of Mexico in order to investigate whether differences in the spectrum of enzyme activities measurable in Arctic and temperate environments are reflected in "downstream" aspects of microbial metabolism (metabolism of monomers and biomass production). Only four of six polysaccharide substrates were hydrolyzed in Smeerenburg Fjord; all were hydrolyzed in the upper water column of the Gulf. These patterns are consistent on an interannual basis. Bacterial protein production was comparable at both locations, but the pathways of glucose utilization differed. Glucose incorporation rate constants were comparatively higher in Svalbard, but glucose respiration rate constants were higher in surface waters of the Gulf. As a result, at the time of sampling ca. 75% of the glucose was incorporated into biomass in Svalbard, but in the northern Gulf of Mexico most of the glucose was respired to CO2. A limited range of enzyme activities is therefore not a sign of a dormant community or one unable to further process substrates resulting from extracellular enzymatic hydrolysis. The ultimate fate of carbohydrates in marine waters, however, is strongly dependent upon the specific capabilities of heterotrophic microbial communities in these disparate environments.

  9. Solid-state cultivation of Grifola frondosa (Dicks: Fr) S.F. Gray biomass and immunostimulatory effects of fungal intra- and extracellular beta-polysaccharides.

    Science.gov (United States)

    Svagelj, Mirjan; Berovic, Marin; Boh, Bojana; Menard, Anja; Simcic, Sasa; Wraber, Branka

    2008-01-01

    Grifola frondosa strain GF3, was cultivated on solid-state substrate consisting of milled whole corn plant (Zea mays) and olive press cake supplemented with mineral additives and olive oil. Maintenance of the moisture content in the solid substrate is of crucial importance. Moistures higher than 70% promote growth of G. frondosa mycelium and polysaccharide production. Four fractions of pure extracellular beta-D-glucans with total mass 127.2mg and four fractions of intracellular polysaccharides with total mass 47.2mg were isolated. Polysaccharides were further separated by ion-exchange, gel and affinity chromatography. Isolated polysaccharide fractions from fungal mycelium proved to induce moderate amounts of TNF-alpha in PBMC cells in vitro. The extent of TNF-alpha induction was up to 322pgmL(-1) at a polysaccharide concentration of 200microgmL(-1) for the intracellular fraction. The TNF-alpha inducing activity is comparable to romurtide, which has been used as a supporting therapy in cancer patients treated with radiotherapy and/or chemotherapy.

  10. THE COMPLEX EXTRACELLULAR POLYSACCHARIDES OF MAINLY CHAIN-FORMING FRESHWATER DIATOM SPECIES FROM EPILITHIC BIOFILMS(1).

    Science.gov (United States)

    Bahulikar, Rahul A; Kroth, Peter G

    2008-12-01

    Diatoms are dominant organisms in phototrophic biofilms in aquatic habitats. They produce copious amounts of extracellular polymeric substances (EPS), which mainly consist of carbohydrates and traces of proteins and glycoproteins. This study focuses on the characterization of EPS from a total of 14 diatoms belonging to the six genera Achnanthes, Cymbella, Fragilaria, Punctastriata, Staurosira, and Pseudostaurosira, all of which were isolated from epilithic biofilms of the littoral zone of Lake Constance. EPS from all isolates were extracted by a sequential extraction procedure resulting in five different fractions. The monosaccharide composition of each fraction was analyzed by HPLC equipped with a pulse amperiometric detector, yielding results similar to those obtained by probing the EPS structures with monomer-specific fluorophore-linked lectins. Significant differences in carbohydrate composition occurred in the different fractions of single isolates. Most of the diatom isolates in our study form chain-like colonies in which the cells are attached to each other by intercellular pads. Here we demonstrate that these pads can be dissolved in hot bicarbonate and that they show a heterogeneous composition of monosaccharides in contrast to other fractions, which mostly were dominated by one or two monosaccharides. Principal component analysis indicates a correlation between carbohydrate composition of EPS fractions and the phylogenetic relationship of the respective species, indicating that EPS analyses under defined culture conditions may support taxonomic analyses.

  11. Roles of Extracellular Polysaccharides and Biofilm Formation in Heavy Metal Resistance of Rhizobia

    Directory of Open Access Journals (Sweden)

    Natalia Nocelli

    2016-05-01

    Full Text Available Bacterial surface components and extracellular compounds, particularly flagella, lipopolysaccharides (LPSs, and exopolysaccharides (EPSs, in combination with environmental signals and quorum-sensing signals, play crucial roles in bacterial autoaggregation, biofilm development, survival, and host colonization. The nitrogen-fixing species Sinorhizobium meliloti (S. meliloti produces two symbiosis-promoting EPSs: succinoglycan (or EPS I and galactoglucan (or EPS II. Studies of the S. meliloti/alfalfa symbiosis model system have revealed numerous biological functions of EPSs, including host specificity, participation in early stages of host plant infection, signaling molecule during plant development, and (most importantly protection from environmental stresses. We evaluated functions of EPSs in bacterial resistance to heavy metals and metalloids, which are known to affect various biological processes. Heavy metal resistance, biofilm production, and co-culture were tested in the context of previous studies by our group. A range of mercury (Hg II and arsenic (As III concentrations were applied to S. meliloti wild type strain and to mutant strains defective in EPS I and EPS II. The EPS production mutants were generally most sensitive to the metals. Our findings suggest that EPSs are necessary for the protection of bacteria from either Hg (II or As (III stress. Previous studies have described a pump in S. meliloti that causes efflux of arsenic from cells to surrounding culture medium, thereby protecting them from this type of chemical stress. The presence of heavy metals or metalloids in culture medium had no apparent effect on formation of biofilm, in contrast to previous reports that biofilm formation helps protect various microorganism species from adverse environmental conditions. In co-culture experiments, EPS-producing heavy metal resistant strains exerted a protective effect on AEPS-non-producing, heavy metal-sensitive strains; a phenomenon

  12. Extracellular Enzyme Activity assay as indicator of soil microbial functional diversity and activity

    DEFF Research Database (Denmark)

    Hendriksen, Niels Bohse; Winding, Anne

    2012-01-01

    Extracellular Enzyme Activity assay as indicator of soil microbial functional diversity and activity Niels Bohse Hendriksen, Anne Winding. Department of Environmental Science, Aarhus University, 4000 Roskilde, Denmark Soil enzymes originate from a variety of organisms, notably fungi and bacteria...... and especially hydrolytic extracellular enzymes are of pivotal importance for decomposition of organic substrates and biogeochemical cycling. Their activity reflects the functional diversity and activity of the microorganisms involved in decomposition processes which are essential processes for soil functioning......, experimental conditions of extraction of enzymes from soils, buffer and pH, substrate concentration, temperature and the necessary controls were optimized and standardized. This has resulted in an optimized standard operating procedure of EEA, which are being tested as an indicator of soil functional diversity...

  13. The role of glucose kinase in carbohydrate utilization and extracellular polysaccharide production in Xanthomonas campestris pathovar campestris.

    Science.gov (United States)

    Lu, Guang-Tao; Yang, Zheng-Jiu; Peng, Fang-Yin; Tan, Yi-Ning; Tang, Yong-Qin; Feng, Jia-Xun; Tang, Dong-Jie; He, Yong-Qiang; Tang, Ji-Liang

    2007-12-01

    The genome of the Xanthomonas campestris pathovar campestris (Xcc) strain 8004 encodes three uncharacterized proteins, XC1166, XC1223 and XC1976, annotated as glucose kinase (Glk) by bioinformatic studies. Here we have investigated the biochemical characteristics and physiological roles of these proteins with particular reference to the synthesis of extracellular polysaccharide (EPS). XC1166, XC1223 and XC1976 were overexpressed as fusion proteins with a His(6) affinity tag and purified by nickel affinity chromatography. The standard Glk activity assay revealed that all three proteins possessed apparent Glk activity, with XC1976-His(6) being the most active; the specific activity values were 1.16x10(6) U mg(-1) for XC1166-His(6), 4.36x10(7) U mg(-1) for XC1223-His(6) and 2.63x10(8) U mg(-1) for XC1976-His(6). TLC analysis showed, however, that only XC1976-His(6) could phosphorylate glucose. Insertional mutants of XC1166, XC1223 and XC1976 were generated using the suicide plasmid pK18mob. Although mutant strains with insertions in XC1166 or XC1223 had Glk activity similar to that of the wild-type strain, the XC1976 mutant had only about 6% of the wild-type activity. Mutation in XC1976 had complex effects on EPS production. In media containing arabinose, glucose, galactose, sucrose or maltose, the XC1976 mutant produced about 40-75% of the wild-type level of EPS, whereas in medium containing fructose, the mutant showed a 30% increase in EPS production compared to the wild-type strain. The XC1976 mutant also showed attenuated virulence on the host plant Chinese radish (Raphanus sativus). The results indicate that XC1976 has the most significant role for the parameters tested.

  14. Identification of an Efflux Transporter LmrB Regulating Stress Response and Extracellular Polysaccharide Synthesis in Streptococcus mutans

    Directory of Open Access Journals (Sweden)

    Jia Liu

    2017-06-01

    Full Text Available Efflux transporters have been implicated in regulating bacterial virulence properties such as resistance to antibiotics, biofilm formation and colonization. The pathogenicity of Streptococcus mutans, the primary etiologic agent of human dental caries, relies on the bacterium’s ability to form biofilms on tooth surface. However, the studies on efflux transporters in S. mutans are scare and the function of these transporters remained to be clarified. In this study, we identified an efflux transporter (LmrB in S. mutans through cloning the lmrB gene into Escherichia coli. Introducing lmrB into E. coli conferred a multidrug-resistant phenotype and resulted in higher EtBr efflux activity which could be suppressed by efflux inhibitor. To explore whether LmrB was involved in S. mutans virulence properties regulation, we constructed the lmrB inactivation mutant and examined the phenotypes of the mutant. It was found that LmrB deficiency resulted in increased IPS storage and prolonged acid production. Enhanced biofilm formation characterized by increased extracellular polysaccharides (EPS production and elevated resistance to hydrogen peroxide and antimicrobials were also observed in lmrB mutant. To gain a better understanding of the global role of LmrB, a transcriptome analysis was performed using lmrB mutant strain. The expression of 107 genes was up- or down-regulated in the lmrB mutant compared with the wild type. Notably, expression of genes in several genomic islands was differentially modulated, such as stress-related GroELS and scnRK, sugar metabolism associated glg operons and msmREFGK transporter. The results presented here indicate that LmrB plays a vital global role in the regulation of several important virulence properties in S. mutans.

  15. Metal affinity enrichment increases the range and depth of proteome identification for extracellular microbial proteins

    Energy Technology Data Exchange (ETDEWEB)

    Wheeler, Korin [Lawrence Livermore National Laboratory (LLNL); Erickson, Brian K [ORNL; Mueller, Ryan [University of California, Berkeley; Singer, Steven [Lawrence Livermore National Laboratory (LLNL); Verberkmoes, Nathan C [ORNL; Hwang, Mona [Lawrence Livermore National Laboratory (LLNL); Thelen, Michael P. [University of California, Berkeley; Hettich, Robert {Bob} L [ORNL

    2012-01-01

    Many key proteins, such as those involved in cellular signaling or transcription, are difficult to measure in microbial proteomic experiments due to the interfering presence of more abundant, dominant proteins. In an effort to enhance the identification of previously undetected proteins, as well as provide a methodology for selective enrichment, we evaluated and optimized immobilized metal affinity chromatography (IMAC) coupled with mass spectrometric characterization of extracellular proteins from an extremophilic microbial community. Seven different metals were tested for IMAC enrichment. The combined results added 20% greater proteomic depth to the extracellular proteome. Although this IMAC enrichment could not be conducted at the physiological pH of the environmental system, this approach did yield a reproducible and specific enrichment of groups of proteins with functions potentially vital to the community, thereby providing a more extensive biochemical characterization. Notably, 40 unknown proteins previously annotated as hypothetical were enriched and identified for the first time. Examples of identified proteins includes a predicted TonB signal sensing protein homologous to other known TonB proteins and a protein with a COXG domain previously identified in many chemolithoautotrophic microbes as having a function in the oxidation of CO.

  16. Nitrogen doped carbon nanoparticles enhanced extracellular electron transfer for high-performance microbial fuel cells anode.

    Science.gov (United States)

    Yu, Yang-Yang; Guo, Chun Xian; Yong, Yang-Chun; Li, Chang Ming; Song, Hao

    2015-12-01

    Nitrogen doped carbon nanoparticles (NDCN) were applied to modify the carbon cloth anodes of microbial fuel cells (MFCs) inoculated with Shewanella oneidensis MR-1, one of the most well-studied exoelectrogens. Experimental results demonstrated that the use of NDCN increased anodic absorption of flavins (i.e., the soluble electron mediator secreted by S. oneidensis MR-1), facilitating shuttle-mediated extracellular electron transfer. In addition, we also found that NDCN enabled enhanced contact-based direct electron transfer via outer-membrane c-type cytochromes. Taken together, the performance of MFCs with the NDCN-modified anode was enormously enhanced, delivering a maximum power density 3.5 times' higher than that of the MFCs without the modification of carbon cloth anodes.

  17. Extracellular Matrix Assembly in Diatoms (Bacillariophyceae) (I. A Model of Adhesives Based on Chemical Characterization and Localization of Polysaccharides from the Marine Diatom Achnanthes longipes and Other Diatoms).

    Science.gov (United States)

    Wustman, B. A.; Gretz, M. R.; Hoagland, K. D.

    1997-04-01

    Extracellular adhesives from the diatoms Achnanthes longipes, Amphora coffeaeformis, Cymbella cistula, and Cymbella mexicana were characterized by monosaccharide and methylation analysis, lectin-fluorescein isothiocyanate localization, and cytochemical staining. Polysaccharide was the major component of adhesives formed during cell motility, synthesis of a basal pad, and/or production of a highly organized shaft. Hot water-insoluble/hot 0.5 M NaHCO3-soluble anionic polysaccharides from A. longipes and A. coffeaeformis adhesives were primarily composed of galactosyl (64-70%) and fucosyl (32-42%) residues. In A. longipes polymers, 2,3-, t-, 3-, and 4-linked/substituted galactosyl, t-, 3-, 4-, and 2-linked fucosyl, and t- and 2-linked glucuronic acid residues predominated. Adhesive polysaccharides from C. cistula were EDTA-soluble, sulfated, consisted of 83% galactosyl (4-, 4,6-, and 3,4-linked/substituted) and 13% xylosyl (t-, 4f/5p-, and 3p-linked/substituted) residues, and contained no uronosyl residues. Ulex europaeus agglutinin uniformly localized [alpha](1,2)-L-fucose units in C. cistula and Achnanthes adhesives formed during motility and in the pads of A. longipes. D-Galactose residues were localized throughout the shafts of C. cistula and capsules of A. coffeaeformis. D-Mannose and/or D-glucose, D-galactose, and [alpha](t)-L-fucose residues were uniformly localized in the outer layers of A. longipes shafts by Cancavalia ensiformis, Abrus precatorius, and Lotus tetragonolobus agglutinin, respectively. A model for diatom cell adhesive structure was developed from chemical characterization, localization, and microscopic observation of extracellular adhesive components formed during the diatom cell-attachment process.

  18. Implications of Extracellular Polymeric Substance Matrices of Microbial Habitats Associated with Coastal Aquaculture Systems

    Directory of Open Access Journals (Sweden)

    Juan Carlos Camacho-Chab

    2016-08-01

    Full Text Available Coastal zones support fisheries that provide food for humans and feed for animals. The decline of fisheries worldwide has fostered the development of aquaculture. Recent research has shown that extracellular polymeric substances (EPS synthesized by microorganisms contribute to sustainable aquaculture production, providing feed to the cultured species, removing waste and contributing to the hygiene of closed systems. As ubiquitous components of coastal microbial habitats at the air–seawater and seawater–sediment interfaces as well as of biofilms and microbial aggregates, EPS mediate deleterious processes that affect the performance and productivity of aquaculture facilities, including biofouling of marine cages, bioaccumulation and transport of pollutants. These biomolecules may also contribute to the persistence of harmful algal blooms (HABs and their impact on cultured species. EPS may also exert a positive influence on aquaculture activity by enhancing the settling of aquaculturally valuable larvae and treating wastes in bioflocculation processes. EPS display properties that may have biotechnological applications in the aquaculture industry as antiviral agents and immunostimulants and as a novel source of antifouling bioproducts.

  19. The role of microbial-produced extracellular polymeric matrix in the formation and survival of biological soil crusts

    Science.gov (United States)

    Rossi, Federico; Adessi, Alessandra; De Philippis, Roberto

    2016-04-01

    Biological soil crusts (BSCs) are complex communities commonly constituting organo-mineral layers in arid and semiarid environment having a major influence on these ecosystems (Belnap and Lange, 2001). They have high tolerance towards a-biotic stresses and fluctuations in moisture, illumination, salinity and nutrients. The plasticity exhibited by BSCs is hugely contributed by the presence of the extracellular polymeric matrix (EPM) that is synthesized by crustal organisms, notably cyanobacteria and microalgae. This polysaccharidic net plays key roles in biofilm relations with the surrounding constrained environment. Notably, EPM concurs in coping with water scarcity, freezing and salt stress; increases biolayers stability against erosion, and is involved in nutrient provision (Rossi and De Philippis, 2015). We conducted several investigations in a research area located in the Inner Mongolian desert (Inner Mongolia, China) where BSCs were induced over different sites through inoculation-based techniques performed in different years. Our studies were aimed at determining the role of EPM in BSC development and survival in such a hyper-arid system. This presentation will report the results concerning the role of EPM in water capture from non-rainfall sources, water maintenance at the topsoil, and in water infiltrability, the latter being a factor with important ecological implications. In additions we investigated the role of the matrix as a source of carbon for the crustal heterotrophs. Furthermore, EPM was extracted with methods optimized in our lab, aiming at removing tightly bound fractions and loosely bound fractions from BSCs having different ages. The fractions were analyzed in terms of monosaccharidic composition, and molecular weight (MW) distribution. We show how the relative amounts of uronic acids increase in the EPM with the age of the crusts, implying advantages for the community-water relations. In addition, we observed significant differences in MW

  20. Competitive adsorption of Reactive Orange 16 and Reactive Brilliant Blue R on polyaniline/bacterial extracellular polysaccharides composite-A novel eco-friendly polymer

    Energy Technology Data Exchange (ETDEWEB)

    Janaki, V. [Department of Chemistry, Periyar University, Salem 636011, Tamil Nadu (India); Vijayaraghavan, K. [Singapore-Delft Water Alliance, National University of Singapore, 117577 (Singapore); Ramasamy, A.K. [Department of Chemistry, Periyar University, Salem 636011, Tamil Nadu (India); Lee, Kui-Jae [Division of Biotechnology, Advanced Institute of Environment and Bioscience, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan 570752 (Korea, Republic of); Oh, Byung-Taek, E-mail: btoh@jbnu.ac.kr [Division of Biotechnology, Advanced Institute of Environment and Bioscience, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan 570752 (Korea, Republic of); Kamala-Kannan, Seralathan, E-mail: kannan@jbnu.ac.kr [Division of Biotechnology, Advanced Institute of Environment and Bioscience, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan 570752 (Korea, Republic of)

    2012-11-30

    Highlights: Black-Right-Pointing-Pointer Competitive adsorption of reactive dyes onto polyaniline/bacterial extracellular polysaccharides composite. Black-Right-Pointing-Pointer The composite have functional groups of both polyaniline and bacterial extracellular polysaccharides. Black-Right-Pointing-Pointer The presence of Reactive Brilliant Blue R diminished the uptake of Reactive Orange 16. Black-Right-Pointing-Pointer Electrostatic interaction was identified as a major mechanism in adsorption process. Black-Right-Pointing-Pointer Reactive Brilliant Blue R and Reactive Orange 16 adsorption was endothermic process. - Abstract: The performance of polyaniline/extracellular polymeric substances (Pn/EPS) composite as an adsorbent to remove the anionic reactive dyes, Reactive Brilliant Blue R (RBBR) and Reactive Orange 16 (RO), was investigated in single and binary systems. The pH{sub pzc} of Pn/EPS composite was calculated as 3.7 through potentiometric mass titration method. Electrostatic interaction between the dye anion and the nitrogen present in the polymer was identified as a major mechanism in adsorption process. Single component isotherms followed the Langmuir model with the maximum adsorption capacity of 0.5775 mmol g{sup -1} for RBBR and 0.4748 mmol g{sup -1} for RO. In binary system, both the reactive dye anions compete with each other and resulted in lower uptake. Binary adsorption data were interpreted well by the Sheindorf-Rehbun-Sheintuch equation as compared to extended Langmuir model with constant interaction factor. Kinetic analysis of single solute followed pseudo-first order model. Thermodynamic studies computed that RBBR and RO adsorption was endothermic, spontaneous, and feasible process.

  1. Extracellular polysaccharide production by a novel osmotolerant marine strain of Alteromonas macleodii and its application towards biomineralization of silver.

    Directory of Open Access Journals (Sweden)

    Ananya Mehta

    Full Text Available The present study demonstrates exopolysaccharide production by an osmotolerant marine isolate and also describes further application of the purified polysaccharide for production of colloidal suspension of silver nanoparticles with narrow size distribution. Phylogenetic analysis based on 16S r RNA gene sequencing revealed close affinity of the isolate to Alteromonas macleodii. Unlike earlier reports, where glucose was used as the carbon source, lactose was found to be the most suitable substrate for polysaccharide production. The strain was capable of producing 23.4 gl(-1 exopolysaccharide with a productivity of 7.8 gl(-1 day(-1 when 15% (w/v lactose was used as carbon source. Furthermore, the purified polysaccharide was able to produce spherical shaped silver nanoparticles of around 70 nm size as characterized by Uv-vis spectroscopy, Dynamic light scattering and Transmission electron microscopy. These observations suggested possible commercial potential of the isolated strain for production of a polysaccharide which has the capability of synthesizing biocompatible metal nanoparticle.

  2. A common theme in extracellular fluids of beetles: extracellular superoxide dismutases crucial for balancing ROS in response to microbial challenge

    Science.gov (United States)

    Gretscher, René R.; Streicher, Priska E.; Strauß, Anja S.; Wielsch, Natalie; Stock, Magdalena; Wang, Ding; Boland, Wilhelm; Burse, Antje

    2016-01-01

    Extracellular Cu/Zn superoxide dismutases (SODs) are critical for balancing the level of reactive oxygen species in the extracellular matrix of eukaryotes. In the present study we have detected constitutive SOD activity in the haemolymph and defensive secretions of different leaf beetle species. Exemplarily, we have chosen the mustard leaf beetle, Phaedon cochleariae, as representative model organism to investigate the role of extracellular SODs in antimicrobial defence. Qualitative and quantitative proteome analyses resulted in the identification of two extracellular Cu/Zn SODs in the haemolymph and one in the defensive secretions of juvenile P. cochleariae. Furthermore, quantitative expression studies indicated fat body tissue and defensive glands as the main synthesis sites of these SODs. Silencing of the two SODs revealed one of them, PcSOD3.1, as the only relevant enzyme facilitating SOD activity in haemolymph and defensive secretions in vivo. Upon challenge with the entomopathogenic fungus, Metarhizium anisopliae, PcSOD3.1-deficient larvae exhibited a significantly higher mortality compared to other SOD-silenced groups. Hence, our results serve as a basis for further research on SOD regulated host-pathogen interactions. In defensive secretions PcSOD3.1-silencing affected neither deterrent production nor activity against fungal growth. Instead, we propose another antifungal mechanism based on MRJP/yellow proteins in the defensive exudates. PMID:27068683

  3. Microbial respiration and kinetics of extracellular enzymes activities through rhizosphere and detritusphere at agricultural site

    Science.gov (United States)

    Löppmann, Sebastian; Blagodatskaya, Evgenia; Kuzyakov, Yakov

    2014-05-01

    detritivore communities in the soil. The kinetics (Km and Vmax) of four extracellular hydrolytic enzymes responsible for C- and phosphorous-cycle (β-glucosidase, β-xylosidase, β-cellobiohydrolase and acid phosphatase), microbial biomass, basal respiration (BR) and substrate-induced respiration (SIR) were measured in rhizosphere, detritusphere and control from 0 - 10 and 10 - 20 cm. The metabolic quotient (qCO2) was calculated as specific indicator for efficiency of microbial substrate utilization. We observed clear differences in enzymes activities at low and high concentrations of substrate. At substrate saturation enzyme activity rates of were significantly higher in rooted plots compared to litter amended plots, whereas at lower concentration no treatment effect could be found. The BR, SIR and qCO2 values were significantly higher at 0 - 10 cm of the planted treatment compared to litter and control plots, revealing a significantly higher respiration at lower efficiency of microbial substrate utilization in the rhizosphere. The Michaelis-Menten constant (Km) decreased with depth, especially for β-glucosidase, acid phosphatase and β-xylosidase, indicating higher substrate affinity of microorganisms in deeper soil and therefore different enzyme systems functioning. The substrate affinity factor (Vmax/Km) increased 2-fold with depth for various enzymes, reflecting a switch of predominantly occurring microbial strategies. Vmax/Km ratio indicated relative domination of zymogenous microbial communities (r-strategists) in 0 - 10 cm depth as compared with 10 - 20 cm depth where the K-strategists dominated.

  4. Sulfated Modification of Extracellular Polysaccharide from Submerged Fermentation of Ganoderma lucidum%发酵灵芝胞外多糖硫酸化修饰

    Institute of Scientific and Technical Information of China (English)

    沈洁; 刘昱均; 张珏

    2015-01-01

    The extracellular polysaccharides from submerged fermentation of Ganoderma lucidum exhibit low anticancer activity. Sulfation of polysaccharides is an important approach to improve anticancer activity of polysaccharides. In addition, the degree of sulfated substitution (DS) is closely related to its biological activities. In order to understand the relationship between DS and sulfated conditions and obtain the sulfated derivatives with different DS, the sulfuric acid esterification reagent and the relationship between the sulfated reaction conditions and DS were further investigated. The results showed that aminosulfonic acid offered milder reaction conditions than that of chlorosulfonic acid. On the other hand, the addition of urea also gave mild reaction conditions and improved DS.%发酵灵芝胞外多糖表现出微弱的抗肿瘤活性,硫酸化是改善多糖抑癌活性的重要途径,而多糖的硫酸化取代度与其生物活性密切相关。为了了解硫酸化取代度与硫酸化条件之间的关系,以获取不同取代度的产品,对发酵灵芝胞外多糖的硫酸化试剂以及硫酸化反应条件与取代度之间的关系进行了较深入研究。实验结果表明,氨基磺酸比氯磺酸的硫酸化过程更加缓和,尿素的加入也有利于缓和反应条件,提高取代度。

  5. Ecophysiological Analysis of Microorganisms in Complex Microbial Systems by Combination of Fluorescence In Situ Hybridization with Extracellular Staining Techniques

    Science.gov (United States)

    Nielsen, Jeppe Lund; Kragelund, Caroline; Nielsen, Per Halkjær

    Ecophysiological analysis and functions of single cells in complex microbial systems can be examined by simple combinations of Fluorescence in situ hybridization (FISH) for identification with various staining techniques targeting functional phenotypes. In this chapter, we describe methods and protocols optimized for the study of extracellular enzymes, surface hydrophobicity and specific surface structures. Although primarily applied to the study of microbes in wastewater treatment (activated sludge and biofilms), the methods may also be used with minor modifications in several other ecosystems.

  6. Relevance of fucose-rich extracellular polysaccharides produced by Rhizobium sullae strains nodulating Hedysarum coronarium l. legumes.

    Science.gov (United States)

    Gharzouli, Razika; Carpéné, Marie-Anne; Couderc, François; Benguedouar, Ammar; Poinsot, Véréna

    2013-03-01

    Specific and complex interactions between soil bacteria, known as rhizobia, and their leguminous host plants result in the development of root nodules. This process implies a complex dialogue between the partners. Rhizobia synthesize different classes of polysaccharides: exopolysaccharides (EPS), Kdo-rich capsular polysaccharides, lipopolysaccharides, and cyclic β-(1,2)-glucans. These polymers are actors of a successful symbiosis with legumes. We focus here on studying the EPS produced by Rhizobium sullae bacteria that nodulate Hedysarum coronarium L., largely distributed in Algeria. We describe the influence of the carbon source on the production and on the composition of EPS produced by R. sullae A6 and RHF strains. High-molecular-weight EPS preserve the bacteria from desiccation. The structural characterization of the EPS produced by R. sullae strains has been performed through sugar analysis by gas chromatography-mass spectrometry. The low-molecular-weight EPS of one strain (RHF) has been totally elucidated using nuclear magnetic resonance and quantitative time-of-flight tandem mass spectrometry analyses. An unusual fucose-rich EPS has been characterized. The presence of this deoxy sugar seems to be related to nodulation capacity.

  7. Physical characterization and diel dynamics of different fractions of extracellular polysaccharides in an axenic culture of a benthic diatom

    NARCIS (Netherlands)

    de Brouwer, J.F.C.; Wolfstein, K.; Stal, L.J.

    2002-01-01

    The excretion of extracellular polymeric substances (EPS) by an axenic culture of the benthic diatom Cylindrotheca closterium was investigated. Two sequential extraction steps proved to be sufficient to remove the bulk of the EPS present. Soluble EPS was recovered by a simple centrifugation step and

  8. Alteration of extracellular enzyme activity and microbial abundance by biochar addition: Implication for carbon sequestration in subtropical mangrove sediment.

    Science.gov (United States)

    Luo, Ling; Gu, Ji-Dong

    2016-11-01

    Biochar has attracted more and more attention due to its essential role in adsorbing pollutants, improving soil fertility, and modifying greenhouse gas emission. However, the influences of biochar on extracellular enzyme activity and microbial abundance are still lack and debatable. Currently, there is no information about the impact of biochar on the function of mangrove ecosystems. Therefore, we explored the effects of biochar on extracellular enzyme activity and microbial abundance in subtropical mangrove sediment, and further estimated the contribution of biochar to C sequestration. In this study, sediments were amended with 0 (control), 0.5, 1.0 and 2.0% of biochar and incubated at 25 °C for 90 days. After incubation, enzyme activities, microbial abundance and the increased percentage of sediment organic C content were determined. Both increase (phenol oxidase and β-glucosidase) and decrease (peroxidase, N-acetyl-glucosaminidase and acid phosphatase) of enzyme activities were observed in biochar treatments, but only peroxidase activity showed statistical significance (at least p biochar addition (at least p biochar treatments were remarkably lower than control (p biochar was found. Additionally, the increased percentage of organic C gradually increased with biochar addition rate, which provided evidence for applying biochar to mitigate climate change. Given the importance of microorganisms and enzyme activities in sediment organic matter decomposition, the increased C sequestration might be explained by the large decrease of microbial abundance and enzyme activity after biochar intervention. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Enhancement of extracellular polymeric substances (EPS) production in Spirulina (Arthrospira sp.) by two-step cultivation process and partial characterization of their polysaccharidic moiety.

    Science.gov (United States)

    Chentir, Imene; Hamdi, Marwa; Doumandji, Amel; HadjSadok, Abelkader; Ouada, Hatem Ben; Nasri, Moncef; Jridi, Mourad

    2017-07-05

    The interactive effects of light intensity and NaCl concentration were investigated for Spirulina two-step cultivation process using Full Factorial Design. In the experiment interval, light intensity had no effect while the NaCl concentration had significant effect on the enhancement of extracellular polymeric substances (EPS) production. Interestingly, results revealed a significant negative interaction between light and NaCl concentration indicating that high NaCl concentration (40gL(-1)) and low light intensity (10μmol photons m(-2)s(-1)) enhanced the EPS production. Under these conditions, EPS production reached a maximum of 1.02gg(-1) of biomass (dry weight), which is 1.67-folds greater than EPS content under optimal growth conditions (10μmol photons m(-2)s(-1), 1gL(-1), 30°C). Desalting and deproteinezation steps of EPS were efficient to obtain polysaccharides (PS) with high carbohydrate (67.3±1.1%), low soluble proteins (5.14±0.32%), ash (5.85±0.71%) and sulfate (2.42±0.12%) contents. Rheological studies of PS at different concentrations (1%, 2.5% and 5%) revealed that the viscosity of the solution increased with the increase of PS concentration. In addition, PS exhibited a non Newtonian shear-thinning nature, a predominant gel-like behavior and a good resistance to consecutive heating-cooling cycles. The adopted process could be, then, a promising and economic strategy to enhance EPS production and extract polysaccharides with interesting rheological properties. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Fermentable non-starch polysaccharides increases the abundance of Bacteroides-Prevotella-Porphyromonas in ileal microbial community of growing pigs.

    Science.gov (United States)

    Ivarsson, E; Roos, S; Liu, H Y; Lindberg, J E

    2014-11-01

    Most plant-origin fiber sources used in pig production contains a mixture of soluble and insoluble non-starch polysaccharides (NSP). The knowledge about effects of these sources of NSP on the gut microbiota and its fermentation products is still scarce. The aim of this study was to investigate effects of feeding diets with native sources of NSP on the ileal and fecal microbial composition and the dietary impact on the concentration of short-chain fatty acids (SCFA) and lactic acid. The experiment comprised four diets and four periods in a change-over design with seven post valve t-cecum cannulated growing pigs. The four diets were balanced to be similar in NSP content and included one of four fiber sources, two diets were rich in pectins, through inclusion of chicory forage (CFO) and sugar beet pulp, and two were rich in arabinoxylan, through inclusion of wheat bran (WB) and grass meal. The gut microbial composition was assessed with terminal restriction fragment (TRF) length polymorphism and the abundance of Lactobacillus spp., Enterobacteriaceae, Bacteroides-Prevotella-Porphyromonas and the β-xylosidase gene, xynB, were assessed with quantitative PCR. The gut microbiota did not cluster based on NSP structure (arabinoxylan or pectin) rather, the effect was to a high degree ingredient specific. In pigs fed diet CFO, three TRFs related to Prevotellaceae together consisted of more than 25% of the fecal microbiota, which is about 3 to 23 times higher (PBacteroides-Prevotella-Porphyromonas. The effect on SCFA was correlated to specific neutral sugars where xylose increased the ileal butyric acid proportion, whereas arabinose increased the fecal butyric acid proportion. Moreover, chicory pectin increased the acetic acid proportion in both ileal digesta and feces.

  11. Extracellular polysaccharides are involved in the attachment of Azospirillum brasilense and Rhizobium leguminosarum to arbuscular mycorrhizal structures

    OpenAIRE

    V Bianciotto; Andreotti, S.; Balestrini, R; Bonfante, P.; Perotto, S

    2009-01-01

    Arbuscular mycorrhizal (AM) fungi, one of the most important component of the soil microbial community, establish physical interactions with naturally occurring and genetically modified bacterial biofertilizers and biopesticides, commonly referred to as plant growth-promoting rhizobacteria (PGPR). We have used a genetic approach to investigate the bacterial components possibly involved in the attachment of two PGPR (Azospirillum and Rhizobium) to AM roots and AM fungal structures. Mutants aff...

  12. Extraction and structural properties of Acanthophora muscoides (Rhodophyceae extracellular matrix sulfated polysaccharides and their effects on coagulation

    Directory of Open Access Journals (Sweden)

    José Ariévilo Gurgel Rodrigues

    2016-06-01

    Full Text Available Acanthophora muscoides (Rhodophyta contains structurally heterogeneous sulfated polysaccharides (Am-SPs with pharmacological importance; however, its matrix SPs composition has not been still extensively investigated. This study sequentially extracted and compared the structural features and the in vitro anticoagulant effects of the Am-SPs. Papain-extraction sequence yielded Am.E-1, Am.E-2 and Am.E-3 containing differences among the relative proportions of sulfate (26.18-33% and hexoses (42.02-60.67% based on chemical analyses. One- (1H and two-dimensions (1H/13C nuclear magnetic resonance experiments showed very complex Am-SPs composed of alternating 4-linked-α-galactopyranosyl units and 3-linked-β-galactopyranosyl units presenting variable sulfation, CH3 substitutions and3,6-anhydro-α-L-galactose units and pyruvated-D-galactose residues, respectively, typical of agarocolloids. Different chromatographic profiles (DEAE-cellulose were observed, with fractions (Am I, Am II and Am III eluted with 0.5, 0.75 and/or 1 M of NaCl, respectively revealing charge density patterns and distinct mobility to heparin by agarose-electrophoresis and, when analyzed by polyacrylamide-electrophoresis, a dispersive migration and similar mobility as chondroitin-6-sulfate for Am I fractions were noted. Regarding the activated partial thromboplastin time test, fractions had no virtually anticoagulation (1.47→3.07 IU mg-1 in comparison with 193 IU mg-1 heparin. Therefore, Am-SPs show significantly lower anticoagulation than heparin.

  13. Polysaccharide capsule and suilysin contribute to extracellular survival of Streptococcus suis co-cultivated with primary porcine phagocytes.

    Science.gov (United States)

    Benga, Laurentiu; Fulde, Marcus; Neis, Christina; Goethe, Ralph; Valentin-Weigand, Peter

    2008-11-25

    Streptococcus suis is a major cause of meningitis, sepsis and arthritis in piglets and a zoonotic agent. Survival in the blood circulation system represents a major step in pathogenesis of S. suis infections. To get further insights into the mechanisms of S. suis survival in the host, we compared a highly virulent S. suis serotype 2 strain with its non-encapsulated and suilysin-deficient mutants in their abilities to resist phagocytosis and killing by polymorphonuclear neutrophils (PMNs) and mononuclear cells. PMNs displayed a higher capacity to take up encapsulated bacteria than mononuclear cells, whereas both cell types internalized efficiently non-encapsulated S. suis. Differentiation of extracellular and intracellular survival of the WT strain revealed that in PMNs the majority of the cell-associated streptococci were intracellular, whereas in mononuclear cells the majority remained attached to the cell surface. S. suis survived mainly extracellularly, since both cells killed intracellular bacteria to a similar extent. As a consequence of different resistance to phagocytosis, only the encapsulated S. suis strains survived co-cultivation with PMNs. Comparison of the WT strain with its encapsulated suilysin-deficient mutant revealed reduced survival of the mutant after co-cultivation with PMNs. Involvement of suilysin in inhibition of phagocytosis was further confirmed by the use of anti-suilysin antibodies and recombinant suilysin. Kinetic experiments with PMNs suggested that reduced survival of the mutant strain was mainly associated with an increased uptake, whilst both strains adhered similarly. Concluding, our results indicate that the capsule and the suilysin play important roles in S. suis survival in the host by interfering with phagocytic uptake.

  14. Influence of extracellular polysaccharides (EPS) produced by two different green unicellular algae on membrane filtration in an algae-based biofuel production process.

    Science.gov (United States)

    Matsumoto, Takaki; Yamamura, Hiroshi; Hayakawa, Jyunpei; Watanabe, Yoshimasa; Harayama, Shigeaki

    2014-01-01

    In the present study, two strains of green algae named S1 and S2, categorized as the same species of Pseudo-coccomyxa ellipsoidea but showing 99% homology, were cultivated under the same conditions and filtrated with a microfiltration membrane. On the basis of the results of the extracellular polysaccharides (EPS) characteristics of these two green algae and the degree of fouling, the influence of these characteristics on the performance of membrane filtration was investigated. There was no difference in the specific growth rate between the S1 and S2 strains; however, large differences were seen in the amount and quality of EPS between S1 and S2. When the S1 and S2 strains were filtered with a membrane, the trend in the increase in transmembrane pressure (TMP) was quite different. The filtration of the S1 strain showed a rapid increase in TMP, whereas the TMP of the filtration of the S2 strain did not increase at all during the operation. This clearly demonstrated that the characteristics of each strain affect the development of membrane fouling. On the basis of the detailed characterization of solved-EPS (s-EPS) and bound-EPS (b-EPS), it was clarified that s-EPS mainly contributed to irreversible fouling for both operations and the biopolymer-like organic matter contained in b-EPS mainly contributed to reversible fouling.

  15. Effect of acemannan, an extracted polysaccharide from Aloe vera, on BMSCs proliferation, differentiation, extracellular matrix synthesis, mineralization, and bone formation in a tooth extraction model.

    Science.gov (United States)

    Boonyagul, Sani; Banlunara, Wijit; Sangvanich, Polkit; Thunyakitpisal, Pasutha

    2014-07-01

    Aloe vera is a traditional wound healing medicine. We hypothesized acemannan, a polysaccharide extracted from Aloe vera gel, could affect bone formation. Primary rat bone marrow stromal cells (BMSCs) were treated with various concentrations of acemannan. New DNA synthesis, VEGF, BMP-2, alkaline phosphatase activity, bone sialoprotein, osteopontin expression, and mineralization were determined by [(3)H] thymidine incorporation assay, ELISA, biochemical assay, western blotting, and Alizarin Red staining, respectively. In an animal study, mandibular right incisors of male Sprague-Dawley rats were extracted and an acemannan treated sponge was placed in the socket. After 1, 2, and 4 weeks, the mandibles were dissected. Bone formation was evaluated by dual-energy X-ray absorptiometry and histopathological examination. The in vitro results revealed acemannan significantly increased BMSC proliferation, VEGF, BMP-2, alkaline phosphatase activity, bone sialoprotein and osteopontin expression, and mineralization. In-vivo results showed acemannan-treated groups had higher bone mineral density and faster bone healing compared with untreated controls. A substantial ingrowth of bone trabeculae was observed in acemannan-treated groups. These data suggest acemannan could function as a bioactive molecule inducing bone formation by stimulating BMSCs proliferation, differentiation into osteoblasts, and extracellular matrix synthesis. Acemannan could be a candidate natural biomaterial for bone regeneration.

  16. Characterization of the rcsA Gene from Pantoea sp. Strain PPE7 and Its Influence on Extracellular Polysaccharide Production and Virulence on Pleurotus eryngii

    Science.gov (United States)

    Kim, Min Keun; Lee, Sun Mi; Seuk, Su Won; Ryu, Jae San; Kim, Hee Dae; Kwon, Jin Hyeuk; Choi, Yong Jo; Yun, Han Dae

    2017-01-01

    RcsA is a positive activator of extracellular polysaccharide (EPS) synthesis in the Enterobacteriaceae. The rcsA gene of the soft rot pathogen Pantoea sp. strain PPE7 in Pleurotus eryngii was cloned by PCR amplification, and its role in EPS synthesis and virulence was investigated. The RcsA protein contains 3 highly conserved domains, and the C-terminal end of the open reading frame shared significant amino acid homology to the helix-turn-helix DNA binding motif of bacterial activator proteins. The inactivation of rcsA by insertional mutagenesis created mutants that had decreased production of EPS compared to the wild-type strain and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. The Pantoea sp. strain PPE7 rcsA gene was shown to strongly affect the formation of the disease symptoms of a mushroom pathogen and to act as the virulence factor to cause soft rot disease in P. eryngii. PMID:28592946

  17. Recent Advances in the Study of Marine Microbial Biofilm: From the Involvement of Quorum Sensing in Its Production up to Biotechnological Application of the Polysaccharide Fractions

    Directory of Open Access Journals (Sweden)

    Paola Di Donato

    2016-05-01

    Full Text Available The present review will explore the most relevant findings on marine microbial biofilm, with particular attention towards its polysaccharide fraction, namely exopolysaccharide (EPS. EPSs of microbial origin are ubiquitous in nature, possess unique properties and can be isolated from the bacteria living in a variety of habitats, including fresh water or marine environments, extreme environments or different soil ecosystems. These biopolymers have many application in the field of biotechnology. Several studies showed that the biofilm formation is closely related to quorum sensing (QS systems, which is a mechanism relying on the production of small molecules defined as “autoinducers” that bacteria release in the surrounding environment where they accumulate. In this review, the involvement of microbial chemical communication, by QS mechanism, in the formation of marine biofilm will also be discussed.

  18. Extracellular polymeric substances play roles in extracellular electron transfer of Shewanella oneidensis MR-1

    DEFF Research Database (Denmark)

    Xiao, Yong; Zhang, En-Hua; Christensen, Hans Erik Mølager

    It is well known that microorganism is surrounded by extracellular polymeric substances (EPS) which include polysaccharides, proteins, glycoproteins, nucleic acids, phospholipids, and humic acids. However, previous studies on microbial extracellular electron transfer (EET) are conducted on cells...... the extraction (Figure 1.a and 1.b). Comparing to cells in control group, MR-1 treated at 38 °C for EPS extraction showed different electrochemical characterizations as revealed by differential pulse voltammetry (Figure 1.c). EPS extracted from MR-1 also was proved to be electrochemically active. The present...

  19. Extracellular Saccharide-Mediated Reduction of Au(3+) to Gold Nanoparticles: New Insights for Heavy Metals Biomineralization on Microbial Surfaces.

    Science.gov (United States)

    Kang, Fuxing; Qu, Xiaolei; Alvarez, Pedro J J; Zhu, Dongqiang

    2017-02-15

    Biomineralization is a critical process controlling the biogeochemical cycling, fate, and potential environmental impacts of heavy metals. Despite the indispensability of extracellular polymeric substances (EPS) to microbial life and their ubiquity in soil and aquatic environments, the role played by EPS in the transformation and biomineralization of heavy metals is not well understood. Here, we used gold ion (Au(3+)) as a model heavy metal ion to quantitatively assess the role of EPS in biomineralization and discern the responsible functional groups. Integrated spectroscopic analyses showed that Au(3+)was readily reduced to zerovalent gold nanoparticles (AuNPs, 2-15 nm in size) in aqueous suspension of Escherichia coli or dissolved EPS extracted from microbes. The majority of AuNPs (95.2%) was formed outside Escherichia coli cells, and the removal of EPS attached to cells pronouncedly suppressed Au(3+) reduction, reflecting the predominance of the extracellular matrix in Au(3+) reduction. XPS, UV-vis, and FTIR analyses corroborated that Au(3+) reduction was mediated by the hemiacetal groups (aldehyde equivalents) of reducing saccharides of EPS. Consistently, the kinetics of AuNP formation obeyed pseudo-second-order reaction kinetics with respect to the concentrations of Au(3+) and the hemiacetal groups in EPS, with minimal dependency on the source of microbial EPS. Our findings indicate a previously overlooked, universally significant contribution of EPS to the reduction, mineralization, and potential detoxification of metal species with high oxidation state.

  20. Extracellular polymeric substances act as transient media in extracellular electron transfer of Shewanella oneidensis MR-1

    DEFF Research Database (Denmark)

    Xiao, Yong; Zhang, Jingdong; Ulstrup, Jens

    without extracting EPS or cells collected from log stage or early-steady stage cultures with little EPS. Therefore, microbial cells are believed in contact directly with each other or electrode. Such attempt apparently ignored the role of EPS in microbial EET, even though many components of EPS......It is well known that microorganism is surrounded by extracellular polymeric substances (EPS) which include polysaccharides, proteins, glycoproteins, nucleic acids, phospholipids, and humic acids. However, previous studies on microbial extracellular electron transfer (EET) are conducted on cells......, such as DNA, humic acids and some proteins, are electrochemically active or semiconductive. Herein, we report experimental evidences of EPS role on EET for Shewanella oneidensis MR-1. Atomic force microscopy clearly showed that the cell surface was cleaned and few EPS could be observed on MR-1 after...

  1. MICROBIAL LIPASES: PRODUCTION OF EXTRACELLULAR LIPASE ENZYME BY ALCALIGENES VISCOSUS (DOGE-1 STRAIN

    Directory of Open Access Journals (Sweden)

    P.Sekhar

    2012-05-01

    Full Text Available Industrially important extracellular lipase enzyme production was explored by utilizingmicrobial strain isolated from dairy effluents. Alcaligenes viscosus DOGE-1 strain isolated from dairywaste waters proved to produce extracellular lipase. Various growth factors were attempted to maximizethe lipase production by this strain. Growth factors like NH4PO4, Peptone, Urea coupled with peptone,KH2PO4, Olive oil and pH were found to be favored the maximum lipase production. This microbialstrain was found to have a high lipolytic activity.

  2. Polysaccharide Degradation

    Science.gov (United States)

    Stone, Bruce A.; Svensson, Birte; Collins, Michelle E.; Rastall, Robert A.

    An overview of current and potential enzymes used to degrade polysaccharides is presented. Such depolymerases are comprised of glycoside hydrolases, glycosyl transferases, phosphorylases and lyases, and their classification, active sites and action patterns are discussed. Additionally, the mechanisms that these enzymes use to cleave glycosidic linkages is reviewed as are inhibitors of depolymerase activity; reagents which react with amino acid residues, glycoside derivatives, transition state inhibitors and proteinaceous inhibitors. The characterization of various enzymes of microbial, animal or plant origin has led to their widespread use in the production of important oligosaccharides which can be incorporated into food stuffs. Sources of polysaccharides of particular interest in this chapter are those from plants and include inulin, dextran, xylan and pectin, as their hydrolysis products are purported to be functional foods in the context of gastrointestinal health. An alternative use of degraded polysaccharides is in the treatment of disease. The possibility exists to treat bacterial exopolysaccharide with lyases from bacteriophage to produce oligosaccharides exhibiting bioactive sequences. Although this area is currently in its infancy the knowledge is available to investigate further.

  3. Biofouling of reverse osmosis membranes: effects of cleaning on biofilm microbial communities, membrane performance, and adherence of extracellular polymeric substances.

    Science.gov (United States)

    Al Ashhab, Ashraf; Sweity, Amer; Bayramoglu, Bihter; Herzberg, Moshe; Gillor, Osnat

    2017-05-01

    Laboratory-scale reverse osmosis (RO) flat-sheet systems were used with two parallel flow cells, one treated with cleaning agents and a control (ie undisturbed). The cleaning efforts increased the affinity of extracellular polymeric substances (EPS) to the RO membrane and altered the biofilm surface structure. Analysis of the membrane biofilm community composition revealed the dominance of Proteobacteria. However, within the phylum Proteobacteria, γ-Proteobacteria dominated the cleaned membrane biofilm, while β-Proteobacteria dominated the control biofilm. The composition of the fungal phyla was also altered by cleaning, with enhancement of Ascomycota and suppression of Basidiomycota. The results suggest that repeated cleaning cycles select for microbial groups that strongly attach to the RO membrane surface by producing rigid and adhesive EPS that hampers membrane performance.

  4. 蛹虫草高产胞外虫草素和虫草多糖的诱变育种%Enhanced production of extracellular cordycepin and polysaccharide in Cordyceps militaris by mutation breeding

    Institute of Scientific and Technical Information of China (English)

    孟泽彬; 文庭池; 康冀川; 康超; 王永江

    2012-01-01

    通过诱变获得高产胞外虫草素和虫草多糖的蛹虫草菌株.采用紫外线诱变(UV)、化学诱变(LiCl)、复合诱变(UV-LiCl) 3种方式对蛹虫草孢子进行诱变;发酵检测存活菌株的胞外虫草素和虫草多糖的含量.结果:以胞外虫草素为指标,3种诱变方式的最大正突变率分别为化学突变(29.2%)>紫外突变(28.6%)>复合诱变(26.5%);以胞外多糖为指标,最大正突变率分别为紫外诱变(35.7%)>复合诱变(33.3%)>化学诱变(27.0%).紫外诱变突变株Z-5-1胞外虫草素产量达0.842g/L,比出发菌株高311%;紫外诱变突变株Z-4-7胞外虫草多糖产量达5.250g/L,比出发菌株高148%.在连续培养5代后,仍具有较好的遗传稳定性.紫外诱变能获得较高的蛹虫草正突变率,同时能获得高产虫草素、虫草多糖的突变株.%The aim was to obtain high-yield strains of extracellular cordycepin and polysaccharide of Cordyceps mititaris. Three kind of mutation way, ultraviolet radiation (UV)、lithium chloride (LiCl) and compound mutagenesis (UV-LiCl) were used to deal with the spores of Cordyceps militaris. Fermenting the survival strains and detecting their content of extracellular cordycepin and cordyceps polysaccharide. The results showed: the maximum positive mutation rate order of extracellular cordycepin was LiCl (29.2%)>UV(28.6%)>UV-LiCl(26.5%) respectively, and that of extracellular polysaccharide was UV(35.7%)>UV-LiCl(33.3%)>LiCl(27.0%). The UV-induced mutant strain Z-5-1 had the highest extracellular cordycepin yield of 0.842g/L, which was 311% higher than that of the original strain. And the UV-induced mutant strain Z-4-7 had the highest extracellular Cordyceps polysaccharide production of 5.250g/L, which was 148% higher than that of the original strain. UV mutagenesis could obtain a higher positive mutation rate for Cordyceps militaris, while could produce good mutant strains with high-yield of extracellular cordycepin and

  5. Microbial Exopolysaccharides: Biosynthesis and Potential Applications

    Directory of Open Access Journals (Sweden)

    K. V. Madhuri

    2014-09-01

    Full Text Available Many bacteria synthesize extracellular polysaccharides (EPSs with commercially significant physiological and therapeutic activities. Microbial polysaccharides have also been reported to have potential therapeutic applications. Recently, much attention has been devoted to the microbial exopolysaccharides (EPSs due to their numerous health benefits.EPSs from lactic acid bacteria are reported to possess antitumor effects, immunostimulatory activity, and the ability to lower blood cholesterol. EPSs also offer an alternative class of biothickeners that are widely used in the food and dairy industries and have been proven to provide strong emulsifying activity, which is important in many food formulations. It is also important to understand the mechanism of microbial biosynthesis of EPSs in order to enhance their production by genetic alterations. The potential applications and the mode of microbial biosynthesis of the EPSs have been presented in this article.

  6. Functions of Exosomes and Microbial Extracellular Vesicles in Allergy and Contact and Delayed-Type Hypersensitivity.

    Science.gov (United States)

    Nazimek, Katarzyna; Bryniarski, Krzysztof; Askenase, Philip W

    2016-01-01

    Extracellular vesicles, such as exosomes, are newly recognized intercellular conveyors of functional molecular mechanisms. Notably, they transfer RNAs and proteins between different cells that can then participate in the complex pathogenesis of allergic and related hypersensitivity responses and disease mechanisms, as described herein. This review highlights this important new appreciation of the in vivo participation of such extracellular vesicles in the interactions between allergy-mediating cells. We take into account paracrine epigenetic exchanges mediated by surrounding stromal cells and the endocrine receipt of exosomes from distant cells via the circulation. Exosomes are natural ancient nanoparticles of life. They are made by all cells and in some form by all species down to fungi and bacteria, and are present in all fluids. Besides a new focus on their role in the transmission of genetic regulation, exosome transfer of allergens was recently shown to induce allergic inflammation. Importantly, regulatory and tolerogenic exosomes can potently inhibit allergy and hypersensitivity responses, usually acting nonspecifically, but can also proceed in an antigen-specific manner due to the coating of the exosome surface with antibodies. Deep analysis of processes mediated by exosomes should result in the development of early diagnostic biomarkers, as well as allergen-specific, preventive and therapeutic strategies. These will likely significantly diminish the risks of current allergen-specific parenteral desensitization procedures, and of the use of systemic immunosuppressive drugs. Since extracellular vesicles are physiological, they can be fashioned for the specific delivery of therapeutic molecular instructions through easily tolerated, noninvasive routes, such as oral ingestion, nasal administration, and perhaps even inhalation. © 2016 S. Karger AG, Basel.

  7. Studies of polysaccharides from three edible species of Nostoc (cyanobacteria) with different colony morphologies : structural characterization and effect on the complement system of polysaccharides from Nostoc commune

    NARCIS (Netherlands)

    Brüll, L.P.; Huang, Z.; Thomas-Oates, J.E.; Smestad-Paulsen, B.; Cohen, E.H.; Michaelsen, T.E.

    2000-01-01

    The cyanobacterium Nostoc commune Vaucher produces quite complex extracellular polysaccharides. The cyanobacterium is nitrogen fixing, and on growing the cyanobacterium in media with and without nitrogen, different types of extracellular polysaccharides were obtained. These were also different from

  8. Characterization and Wash Performance Analysis of Microbial Extracellular Enzymes from East Calcutta Wetland in India

    Directory of Open Access Journals (Sweden)

    Ramesh Malathu

    2008-01-01

    Full Text Available Extracellular protease from a novel bacterial isolate showing maximum similarity of 98.22% with Microbacterium luteolum was obtained from East Calcutta Wetland, India. It showed compatibility with commercial detergents. The enzyme retains more than 60% of its activity between 6.0 to 10.5 pH. The maximum activity is at pH 7.5 with 71% activity at pH 10.0 and 10.5. The protease retained its activity between 4 to 60°C with maximum activity at 30°C and a residual activity of 74.4% at 60°C after overnight incubation. It was completely inhibited by 5mM PMSF pointing towards the presence of serine group of protease. Its inhibition by EDTA indicates the involvement of metal cations in its catalytic activity. It is not effected by Cu+2, partially inhibited by Pb+2 and Ni+2, while completely inhibited by Co+1, Cr+6, Zn+1, Al+3, Ag+3 and Hg+2. Strong reducing agents like β- merceptoethanol and oxidants like bleach and hydrogen peroxide inactivate the enzyme. The enzyme retains 88% of its activity on being mixed with commercially available detergents while it is inactivated by non-ionic Triton X 100. Its efficiency as an additive with detergent in terms of cleaning stains like grease, burnt mobil, vegetable curry and blood was found to be satisfactory. It could enhance the quality of washing as additive in case of all the ten detergents that were tried. The protease alone was also capable of cleaning but the detergent additive mixture could work better. The enzyme was found to work efficiently on different colors as well as on fabric. On mixing with detergent it was found to retain activity up to 2 months and there after, there was a drop in efficiency of washing. The bacterial cells were immobilized in calcium alginate and the released enzyme was found to be equally effective. Market surveys were carried out and the satisfactory result prompted the use of another additive (extracellular lipase obtained from yet another bacterial strain from East

  9. Interaction between extracellular matrix molecules and microbial pathogens: the missing link in autoimmunity?

    Directory of Open Access Journals (Sweden)

    Nidhi eSofat

    2015-01-01

    Full Text Available Rheumatoid arthritis (RA is an autoimmune disease characterised by inflammation, tissue rebuilding and fibrosis. Inability by the body to regulate inflammation effectively is one of the hallmarks of RA. Interactions between the external environment and the human host play an important role in the development of autoimmunity. In RA, the observation of anti-cyclic citrullinated peptide antibodies (ACPA to autoantigens is well recognised. Citrullination is a post-translational modification mediated by peptidyl arginine deiminases (PADs, which exist in both mammalian and bacterial forms. Previous studies have shown how proteins expressed in the human extracellular matrix (ECM acquire properties of damage-associated molecular patterns (DAMPs in RA and include collagens, tenascin-C and fibronectin. ECM DAMPs can further potentiate tissue damage in RA. Recent work has shown that citrullination in RA occurs at mucosal sites, including the oral cavity and lung. Mucosal sites have been linked with bacterial infection e.g. periodontal disease, where exogenous pathogens are implicated in the development of autoimmunity via an infectious trigger. Here we explore how mucosal surfaces exposed to bacteria could trigger autoimmunity in RA.

  10. 具鞘微鞘藻胞外多糖抗紫外辐射活性研究%Research on anti-ultraviolet radiation activity of extracellular polysaccharide from Microcoleus vaginatus Gom

    Institute of Scientific and Technical Information of China (English)

    唐倩; 周楠; 唐东山; 曾铁兵; 朱洪; 张晓文

    2015-01-01

    This paper studied the protective effect of extracellular polysaccharide from Microcoleus vaginatus Gom. for E. coli under UV radiation. In the experiment,we prepared bacterial suspension by mixing E. coli with polysaccharide solution,exposed them to ultraviolet radiation,and cultivated for several days and counted the number of E. coli,then computed UV survival rate of E. coli as well as anti ultraviolet radiation rate of polysaccharide. The results showed that,the UV survival rate of E. coli under 15W UV light irradiation conditions was higher than that under the 20W irradiation. Ir-radiated by 20W UV lamp for 40 seconds,the polysaccharide solution of 400 mg/L can achieved 41% of anti-radiation rate,which was 1. 71 times and 1. 47 times that of polysaccharide solution of 200mg/L and 800 mg/L respectively. While irradiated by 15W UV lamp irradiation for 40s,the anti ultraviolet radiation rate was 29% for polysaccharide solution of 400 mg/L,which was 1. 38 times and 1. 32 times that of solution of 200 mg/L and 800 mg/L. The results implied that extracellular polysaccharide from Microcoleus vaginatus Gom. had a certain degree of reinforcing effect for theanti UV radiation of E. coli ,and polysaccharide solution of 400 mg/L worked better than that of oth-er concentrations.%主要研究具鞘微鞘藻胞外多糖在紫外辐射下对大肠埃希氏菌的保护作用,将大肠埃希氏菌与多糖溶液混合制成菌悬液,再分别进行紫外辐射,培养后计数并计算大肠埃希氏菌紫外存活率及多糖抗紫外辐射率。结果表明:15 W紫外灯照射条件下大肠埃希氏菌存活率略高于20 W紫外灯照射条件下大肠埃希氏菌存活率。20 W紫外灯辐射40 s条件下,400 mg/L的多糖溶液的抗紫外辐射率最高达41%,为200 mg/L、800 mg/L多糖溶液抗辐射率的1.71倍和1.47倍;15 W紫外灯辐射40 s条件下,400 mg/L的多糖溶液的抗紫外辐射率为29%,为200 mg/L、800 mg

  11. Respiratory health effects of indoor microbial exposure; a contribution to the development of exposure assessment methods.

    NARCIS (Netherlands)

    Douwes, J.

    1998-01-01

    The studies described in this thesis validated an existing method to measure bacterial endotoxin and explored new methods for measuring mold components in the environment, particularlyβ(1→3)-glucans and extracellular polysaccharides (EPS). These assays were used to study levels of microbial componen

  12. Isolation, chemical characteristics and immunity activity of an extracellular polysaccharide EPSⅠ isolated from Antarctic bacterium Pseudoalteromonas sp. S-15-13

    Institute of Scientific and Technical Information of China (English)

    Li Jiang; Chen Kaoshan; Sun Xiuqin; Song Jinping; Li Guangyou

    2007-01-01

    A new extracelluar polysaccharide (EPS) was isolated and purified from Antarctic bacterium S-15-13, identified as Pseudoalteromonas sp. After being separated and purified by DEAE-Sephadex A-50 ionexchange and Sephadex G-100 gel chromatography, two mains fractions (EPSⅠ and EPSⅡ ) were obtained. EPSⅠ was composed of mannose, glucose and galactose with a molecular weight of 23kDa and EPSⅡ was composed of mannose only with a molecular weight of 62kDa. The effect of the polysaccharide EPSⅠ on the cellular immune response of mice was investigated. Results demonstrated that EPSⅠ could markedly facilitate lymphocyte proliferation, and might be a strong immunomodulator.

  13. Trace heavy metal ions promoted extracellular electron transfer and power generation by Shewanella in microbial fuel cells.

    Science.gov (United States)

    Xu, Yu-Shang; Zheng, Tao; Yong, Xiao-Yu; Zhai, Dan-Dan; Si, Rong-Wei; Li, Bing; Yu, Yang-Yang; Yong, Yang-Chun

    2016-07-01

    Although microbial fuel cells (MFCs) is considered as one of the most promising technology for renewable energy harvesting, low power output still accounts one of the bottlenecks and limits its further development. In this work, it is found that Cu(2+) (0.1μgL(-1)-0.1mgL(-1)) or Cd(2+) (0.1μgL(-1)-1mgL(-1)) significantly improve the electricity generation in MFCs. The maximum power output achieved with trace level of Cu(2+) (∼6nM) or Cd(2+) (∼5nM) is 1.3 times and 1.6 times higher than that of the control, respectively. Further analysis verifies that addition of Cu(2+) or Cd(2+) effectively improves riboflavin production and bacteria attachment on the electrode, which enhances bacterial extracellular electron transfer (EET) in MFCs. These results unveil the mechanism for power output enhancement by Cu(2+) or Cd(2+) addition, and suggest that metal ion addition should be a promising strategy to enhance EET as well as power generation of MFCs.

  14. 双孢菇深层发酵培养基的响应面优化%Optimization of submerged fermentation medium of Agaricus bisporus for extracellular polysaccharide production by response surface analysis

    Institute of Scientific and Technical Information of China (English)

    毛勇; 毛健; 李华钟; 孟祥勇

    2013-01-01

    The effect of carbon sources,nitrogen source and mineral salt on extracellular polysaccharide production by submerged fermentation medium of Agaricus bisporus had been studied. On the base of single factors experiments,response surface analysis was applied to optimize the submerged fermentation medium of Agaricus bisporus for extracellular polysaccharide production. The quadratic regression analysis was applied to get the optimal level of main factors,and optimal quality concentrations of the variables were obtained as follows:glucose 35.7g/L,KH2PO4 2.1g/L and peptone 3.1g/L. Under these optimal conditions,the predicted and experimental production of extracellular polysaccharide was high up to 1.86g/L and 1.87g/L,respectively.%研究了碳源、氮源、无机盐对双孢菇胞外多糖产量的影响.在单因素实验的基础上,采用响应面实验设计对双孢菇(Agaricus bisporus)深层发酵生产胞外多糖的培养基进行了优化,并建立了葡萄糖、KH2PO4、蛋白胨变化的二次回归方程,探讨了各因子对胞外多糖产量的影响.最终确定适宜的培养基条件为葡萄糖35.7g/L,KH2PO42.1g/L,蛋白胨3.1g/L;在此条件下可得到胞外多糖的最大产量,预测值为1.86g/L,对实验结果进行验证,得到胞外多糖的产量为1.87g/L.

  15. 绞股蓝内生真菌产胞外多糖发酵条件的优化%Optimization of Fermentation Conditions of Extracellular Polysaccharide Produced by Endophytic Fungi from Gynostemma pentaphyllum

    Institute of Scientific and Technical Information of China (English)

    李传民; 曹健; 张慧茹; 兰亚莉

    2016-01-01

    Extracellular polysaccharide produced by endophytic fungi JY25 from the medicinal plants Gy-nostemma pentaphyllum had anti-cancer,anti-oxidation and other bioactive effects. The fermentation condi-tions were optimized to obtain the highest EPS. The incubation time,inoculum dose,carbon sources and nitrogen sources were optimized by single factor test and the orthogonal test with the yield of extracellular polysaccharide as the test aim. The optimized results of single factor test showed that the optimum culture cycle was 8 d,the optimum inoculum dose was 7%,the optimum carbon source was glucose,and the opti-mum nitrogen source was yeast powder. The optimized results of orthogonal test indicated that the optimum culture cycle was 8 d,the optimum inoculum dose was 5%,the optimum application rate of glucose was 40 g/L,and the optimum application rate of yeast powder was 5 g/L,under the above conditions the yield of extracellular polysaccharide reached 271. 40 mg/L.%药用植物绞股蓝内生真菌JY25胞外多糖具有抗癌、抗氧化等多种活性作用,对其发酵条件进行优化,以期达到高产胞外多糖的效果. 以胞外多糖产量为检验指标,采用单因素和正交试验对培养时间、接种量、碳源、氮源进行优化. 单因素试验结果表明,绞股蓝内生真菌JY25产胞外多糖的最佳培养周期为8 d,最佳接种量为7%,最佳碳源为葡萄糖,最佳氮源为酵母粉;正交试验优化后发现,产胞外多糖的最佳发酵条件为培养时间8 d、接种量5%、葡萄糖添加量40 g/L、酵母粉添加量5 g/L,在此条件下胞外多糖的产量达到271. 40 mg/L.

  16. Effect of Different Metal Ions on Mycelium Growth and Extracellular Polysaccharide in Medicinal Mushroom Phellinus igniarius%金属离子对桑黄菌丝体及胞外多糖含量的影响

    Institute of Scientific and Technical Information of China (English)

    高慧娟; 刘雨晴; 李娟辉; 董瑞丽; 王晓琴; 贺旭阳; 张万恒; 张芬琴

    2013-01-01

      筛选出促进桑黄菌丝体生物量和胞外多糖的金属离子。通过测定平板上菌丝体生长速度,液体发酵的菌丝体干重和胞外多糖含量,确定金属离子对桑黄生长的影响。在固体平板培养中,Mg2+、Fe2+、Ca2+对桑黄的生长速度具有明显的促进作用,Na+对菌丝生长速度无明显的促进作用,而在液体摇瓶发酵中,1、3 mg/mL Mg2+,0.9 mg/mL Fe2+,3、5、7 mg/mL Ca2+以及3 mg/mL Na+增加菌丝体干重量,较空白对照差异显著,是空白对照的5倍~8倍。在3 mg/mL K+和Na+,7 mg/mL Ca2+,0.7 mg/mL Fe2+,0.1 mg/mL Zn2+的培养基中桑黄分泌的胞外粗多糖含量分别为0.4760、0.7105、0.5200、0.4516和0.6820 g/100 mL,较空白对照显著差异。一定浓度的金属离子对桑黄菌丝体的生长和胞外多糖分泌有一定的促进作用。%To screen the different mineral ions for promoting the growth of mycelia and crude extracellular polysaccharide of hellinus igniarius. Impact of metal ions on the Santo effect on the growth of P. linteus was identified by determination of growth rate of Mycelia in solid surface culture method in medium with different metal ions, mycelia dry weight and crude extracellular polysaccharide obtained in shake -flsak fermentation. The results show that the medium with Mg2+,Fe2+,Ca2+could obviously promote the growth of the mycelia of P. inteus on plate cultivation, while the Na+has no obvious role.In medium with 1 mg/mL and 3 mg/mL Mg2+, 0.9 mg/mL Fe2+, 3,5 mg/mL and 7mg/mL Ca2+,and 3 mg/mLNa+,the dry weight of mycelia were 5-8 folds than that of control medium,In medium with 3 mg/mL K+and Na+, 7mg/mL Ca2+,0.7 mg/mL Fe2+and 0.1 mg/mL Zn2+, the contains of extracellular polysaccharide secreted by P.inteus were 0.476 0, 0.710 5, 0.520 0, 0.451 6 g/100 mL and 0.6820 g/100 mL,respecrively, the difference was significient compared with control medium. The different metal ions had the role of promoting the growth of

  17. Influence of cereal non-starch polysaccharides on ileo-caecal and rectal microbial populations in growing pigs

    DEFF Research Database (Denmark)

    Høgberg, Ann; Lindberg, Jan; Leser, Thomas;

    2004-01-01

    were collected from the ileum, via intestinal post valve T-caecum (PVTC) cannulas surgically inserted at the ileo-caecal ostium, and from the rectum. The total microbial flora of the ileal samples were analysed for by defining base pair length with terminal restriction fraction length polymorphism (T......-RFLP). The microbial diversity of the coliform flora of the ileal and rectal samples were defined by biochemical fingerprinting. It was observed that many terminal restriction fragments (TRFs) disappeared when new diets were introduced and that some characteristic TRFs were found in the high and low NSP diets......, respectively. Both the total gut microflora and the coliform flora were influenced by the dietary NSP content....

  18. Influence of cereal non-starch polysaccharides on ileo-caecal and rectal microbial populations in growing pigs

    DEFF Research Database (Denmark)

    Høgberg, Ann; Lindberg, Jan; Leser, Thomas

    2004-01-01

    -RFLP). The microbial diversity of the coliform flora of the ileal and rectal samples were defined by biochemical fingerprinting. It was observed that many terminal restriction fragments (TRFs) disappeared when new diets were introduced and that some characteristic TRFs were found in the high and low NSP diets......, respectively. Both the total gut microflora and the coliform flora were influenced by the dietary NSP content....

  19. Study on Jerusalem artichoke Pickle Fermented Directly by LactobaciUus plantarum Producing Higher Extracellular Polysaccharide%高产胞外多糖的植物乳杆菌直投发酵菊芋泡菜的研究

    Institute of Scientific and Technical Information of China (English)

    崔树茂; 郭钦; 刘崇万; 董英

    2011-01-01

    采用自主研发的高产胞外多糖的植物乳杆菌直投式泡菜发酵剂制备菊芋泡菜,研究其发酵过程中泡菜液pH、总酸、活菌数、维生素c、氨基酸态氮和可溶性固形物含量的变化,同时探讨菊芋主要成分菊粉对植物乳杆菌生长的影响,并测定该菌胞外多糖的产量。结果表明,菊粉可促进植物乳杆菌的生长,37℃培养24h,该菌胞外多糖产量高达471mg/L。与自然发酵相比,直投发酵的菊芋泡菜各项指标均优于自然发酵,其泡菜液pH达到3.46、总酸含量达到0.36%,活菌数保持在4.3×10^8CFU/mL、维生素C含量为16.92%、氨基酸态氮含量为0.033%、可溶性固形物含量达到15%。得到的菊芋泡菜酸度适宜,呈诱人乳白色,活菌含量高,且富含植物乳杆菌胞外多糖,为老少皆宜的益生泡菜产品。%Jerusalem artichoke polysaccharidc and extraccllular polysaccharide of lactic acid bacteria have different functional properties which are probiotic. In this paper, a Lactobacillus plantarum with high EPS-produclng capacity was selected for direct vat set (DVS) culture starter and prepared for Jerusalem artichoke pickles. The changes of the pH, total acid, viable count, vitamin C, amino acid nitrogen and soluble solids content in the pickle liquid were studied, while the effect of inulin which is the main component of artichoke on growth of Lactobacillus plantarum was studied. The yield of extracellular polysaccharide produced by the strain was determined. The results showed that inulin could promote the growth of Lactobacillus plantarum, and the yield of extracellular polysaccharide was up to 471 mg/L after 24h under the temperature of 37℃. Various indexes of Jerusalem artichoke pickle direct investment indicators were better than natural fermentation. The pH was up to 3.46, total acid content was 0.36% , the number of viable cells remained at 4.3×10^8 CFU/mL, content of

  20. Characterization of the wzc gene from Pantoea sp. strain PPE7 and its influence on extracellular polysaccharide production and virulence on Pleurotus eryngii.

    Science.gov (United States)

    Kim, Min Keun; Lee, Young Han; Kim, Hyeran; Lee, Jeongyeo; Ryu, Jae San

    2015-01-01

    To characterize of the pathogenicity gene from the soft rot pathogen Pantoea sp. PPE7 in Pleurotus eryngii, we constructed over 10,000 kanamycin-resistant transposon mutants of Pantoea sp. strain PPE7 by transposon mutagenesis. One mutant, Pantoea sp. NPPE9535, did not cause a soft rot disease on Pleurotus eryngii was confirmed by the pathogenicity test. The transposon was inserted into the wzc gene and the disruption of the wzc gene resulted in the reduction of polysaccharide production and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. Analysis of the hydropathic profile of this protein indicated that it is composed of two main domains: an N-terminal domain including two transmembrane α-helices and a C-terminal cytoplasmic domain consisting of a tyrosine-rich region. Comparative analysis indicated that the amino acid sequence of Wzc is similar to that of a number of proteins involved in the synthesis or export of polysaccharides in other bacterial species. Purified GST-Wzc was found to affect the phosphorylation of tyrosine residue in vivo. These results showed that the wzc gene might play an important role in the virulence of Pantoea sp. strain PPE7 in P. eryngii.

  1. Microbial products (biosurfactant and extracellular chromate reductase) of marine microorganism are the potential agents reduce the oxidative stress induced by toxic heavy metals.

    Science.gov (United States)

    Gnanamani, A; Kavitha, V; Radhakrishnan, N; Suseela Rajakumar, G; Sekaran, G; Mandal, A B

    2010-09-01

    The present study demonstrates hexavalent chromium reduction and trivalent chromium tolerance behavior of marine Bacillus sp., MTCC 5514 through its extracellular enzyme reductase and biosurfactants production. The isolate reduces 10-2000 mg/L of hexavalent chromium to trivalent chromium with in 24-96 h respectively and the release of extracellular chromium reductase, found responsible for the reduction. Upon reduction, the concentration of trivalent chromium in the medium found comparatively less. Experimental results reveal, biosurfactants activity found responsible for the less concentration of Cr(III). Hypothetically, trivalent chromium upon formation get entrapped in the micelle of biosurfactants, prevents microbial cells from exposure towards trivalent chromium. Thus, the chosen isolate exhibit tolerance and growth with the increasing concentration of chromium.

  2. Extracellular polymers of acid streamers from pyritic mines

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, D.B.; Kelso, W.I.

    1981-01-01

    Extracellular polymers (slimes) extracted from acid streamers found in three disused North Wales mines were found to be a mixture of polysaccharides and RNA. The polymers exist as microfibrils synthesised by viable members of the acid streamer microbial community. Acid streamers from three mines, and from different zones in one of the mines, were shown to contain similar polymers, although the ratio of monomers varied from site to site. Monosaccharides identified in acid hydrolysates of slimes were glucose, galactose, mannose, ribose, xylose, arabinose, rhamnose and fucose.

  3. Controls of Polysaccharide Chemistry on the Kinetics and Thermodynamics of Heterogeneous Calcium Carbonate Nucleation

    Science.gov (United States)

    Giuffre, A. J.; Han, N.; Dove, P. M.

    2011-12-01

    Polysaccharide fibrils control the orientation of calcium carbonate (CaCO3) biominerals. Good examples are found in the multilayered extracellular mucilaginous sheath of green algae and cyanobacteria and in specialized vesicles inside coccolithophorids. More complex organisms such as arthropods and mollusks form biomineralized exoskeletons and shells that consist of insoluble polysaccharides and soluble acid-rich proteins. In these structures, CaCO3 mineral orientation occurs along fibers of the polysaccharide chitin. This raises the question of whether polysaccharide chemistry has specific roles in directing biomineralization. The last three decades of research show that acidic proteins influence CaCO3 polymorph selection, crystallographic orientation, and nucleation and growth rates but little is known about the function of polysaccharides. In fact, polysaccharides are long considered an inert component of organic frameworks. In this experimental investigation, we test the hypothesis that polysaccharides have chemistry-specific influences on calcification by measuring the kinetics of calcite nucleation onto three types of polysaccharide films under controlled solution compositions. Characterized polysaccharides of simple repeating monomer sequences were chosen as model compounds to represent the major carbohydrates seen in microbial and calcifying environments: 1) alginic acid with carboxyl groups, 2) hyaluronic acid with alternating carboxyl and acetylamine groups, and 3) chitosan with amine and acetylamine groups. Biosubstrates were prepared by electrodeposition of these compounds as thin gel-like films onto gold-coated silicon wafers. Using a flow-through cell, heterogeneous nucleation rates of calcite were measured for a suite of supersaturation conditions. These rate data were compared to similar measurements for carboxyl- and hydroxyl-terminated self-assembled monolayers. Calcite nucleation rates onto the three polysaccharides vary by a factor of 400x

  4. Impedance Spectroscopy as a Tool for Non-Intrusive Detection of Extracellular Mediators in Microbial Fuel Cells

    Science.gov (United States)

    2009-12-01

    1):78. Heijnen JJ. 1999. Bioenergetics of microbial growth. In: Flickiger MC, Drew SW, editors. Bioprocess technology: Fermentation , biocatalysis and...Bug juice : Harvesting electricity with microorganisms. Nature Rev Microbiol 4:497. Manohar AK, Bretschger O, Nealson KH, Mansfeld F. 2008. The use of

  5. Competitive adsorption of Reactive Orange 16 and Reactive Brilliant Blue R on polyaniline/bacterial extracellular polysaccharides composite--a novel eco-friendly polymer.

    Science.gov (United States)

    Janaki, V; Vijayaraghavan, K; Ramasamy, A K; Lee, Kui-Jae; Oh, Byung-Taek; Kamala-Kannan, Seralathan

    2012-11-30

    The performance of polyaniline/extracellular polymeric substances (Pn/EPS) composite as an adsorbent to remove the anionic reactive dyes, Reactive Brilliant Blue R (RBBR) and Reactive Orange 16 (RO), was investigated in single and binary systems. The pH(pzc) of Pn/EPS composite was calculated as 3.7 through potentiometric mass titration method. Electrostatic interaction between the dye anion and the nitrogen present in the polymer was identified as a major mechanism in adsorption process. Single component isotherms followed the Langmuir model with the maximum adsorption capacity of 0.5775 mmol g(-1) for RBBR and 0.4748 mmol g(-1) for RO. In binary system, both the reactive dye anions compete with each other and resulted in lower uptake. Binary adsorption data were interpreted well by the Sheindorf-Rehbun-Sheintuch equation as compared to extended Langmuir model with constant interaction factor. Kinetic analysis of single solute followed pseudo-first order model. Thermodynamic studies computed that RBBR and RO adsorption was endothermic, spontaneous, and feasible process.

  6. Characterization, Microbial Community Structure, and Pathogen Occurrence in Urban Faucet Biofilms in South China

    Directory of Open Access Journals (Sweden)

    Huirong Lin

    2015-01-01

    Full Text Available The composition and microbial community structure of the drinking water system biofilms were investigated using microstructure analysis and 454 pyrosequencing technique in Xiamen city, southeast of China. SEM (scanning electron microscope results showed different features of biofilm morphology in different fields of PVC pipe. Extracellular matrix material and sparse populations of bacteria (mainly rod-shaped and coccoid were observed. CLSM (confocal laser scanning microscope revealed different distributions of attached cells, extracellular proteins, α-polysaccharides, and β-polysaccharides. The biofilms had complex bacterial compositions. Differences in bacteria diversity and composition from different tap materials and ages were observed. Proteobacteria was the common and predominant group in all biofilms samples. Some potential pathogens (Legionellales, Enterobacteriales, Chromatiales, and Pseudomonadales and corrosive microorganisms were also found in the biofilms. This study provides the information of characterization and visualization of the drinking water biofilms matrix, as well as the microbial community structure and opportunistic pathogens occurrence.

  7. NMR and MALDI-TOF MS based characterization of exopolysaccharides in anaerobic microbial aggregates from full-scale reactors

    KAUST Repository

    Gonzalez-Gil, Graciela

    2015-09-22

    Anaerobic granular sludge is composed of multispecies microbial aggregates embedded in a matrix of extracellular polymeric substances (EPS). Here we characterized the chemical fingerprint of the polysaccharide fraction of EPS in anaerobic granules obtained from full-scale reactors treating different types of wastewater. Nuclear magnetic resonance (NMR) signals of the polysaccharide region from the granules were very complex, likely as a result of the diverse microbial population in the granules. Using nonmetric multidimensional scaling (NMDS), the 1H NMR signals of reference polysaccharides (gellan, xanthan, alginate) and those of the anaerobic granules revealed that there were similarities between the polysaccharides extracted from granules and the reference polysaccharide alginate. Further analysis of the exopolysaccharides from anaerobic granules, and reference polysaccharides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) revealed that exopolysaccharides from two of the anaerobic granular sludges studied exhibited spectra similar to that of alginate. The presence of sequences related to the synthesis of alginate was confirmed in the metagenomes of the granules. Collectively these results suggest that alginate-like exopolysaccharides are constituents of the EPS matrix in anaerobic granular sludge treating different industrial wastewater. This finding expands the engineered environments where alginate has been found as EPS constituent of microbial aggregates.

  8. Characterization of extracellular polymeric substances and microbial diversity in anaerobic co-digestion reactor treated sewage sludge with fat, oil, grease.

    Science.gov (United States)

    Yang, Zhao-Hui; Xu, Rui; Zheng, Yue; Chen, Ting; Zhao, Li-Jun; Li, Min

    2016-07-01

    Performance of co-digesters, treated of sewage sludge (SS) with fat, oil and grease (FOG), were conducted semi-continuously in two mesophilic reactors over 180days. Compared with SS mono-digestion, biogas production and TS removal efficiency of co-digestion were significantly enhanced up to 35% and 26% by adding upper limit FOG (60% on VS). Enhancement in co-digestion performance was also stimulated by the release of extracellular polymeric substances (EPS), which was increased 40% in both loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) than that of mono-digester. Based on high-throughput sequencing (HTS), analysis of microbial 16S rRNA gene comprehensively revealed the dynamic change of microbial community. Results showed that both bacterial and archaeal undergone an apparent succession with FOG addition, and large amount of consortium like Methanosaeta and N09 were involved in the process. Redundancy analysis showed the acetoclastic genera Methanosaeta distinctly related with biogas production and EPS degradation.

  9. Polymeric and compositional properties of novel extracellular microbial polyglucosamine biopolymer from new strain of citrobacter sp. BL-4.

    Science.gov (United States)

    Kim, Lin-Su; Hong, Soo-Jung; Son, Mi-Kyung; Lee, Yong-Hyun

    2006-02-01

    A novel polyglucosamine polymer, PGB-2, was produced extracellularly from a new strain Citrobacter sp. BL-4 using pH-stat fed batch cultivation. It was composed of 97.3% glucosamine and 2.7% rhamnose; its average molecular weight, solubility in 2% acetic acid and viscosity were 20 kDa, 5 g l(-1) and 2.9 cps, respectively. FT-IR and 1H NMR spectra of PGB-2 revealed a close identity with chitosan from crab shells.

  10. Impact of fire, landscape position, aspect, and soil depth on microbial extracellular enzyme activities in the Jemez River Basin Critical Zone Observatory.

    Science.gov (United States)

    Fairbanks, D.; Murphy, M. A.; Frost, G.; Chorover, J.; Gallery, R. E.; Rich, V. I.

    2014-12-01

    Fire frequency and severity are increasing across the western US, and post-fire recovery and effects on critical zone structure are not fully understood. Resident microbiota (bacteria and fungi) transform the majority of carbon in ecosystems, and the structure of these communities influence seedling establishment and the trajectory of vegetative recovery as well as biogeochemical cycling. We surveyed changes in microbial composition and activity after wildfire to better understand soil microbial resilience and fire ecology. Specifically, we assessed potential extracellular enzyme activities in response to fire severity across landscape position and aspect. We sampled 18 days after containment of the June 2013 Thompson Ridge Fire in the Jemez River Basin Critical Zone Observatory, across a gradient of burn severities in a mixed-conifer zero order basin. We subsampled six depths through the surface soil profile and measured potential activities of seven hydrolytic enzymes using established fluorometric techniques. Four of these enzymes hydrolyze C-rich substrates (β-glucosidase [BG], β-D-cellubiosidase [CB], xylosidase [XYL], and α-glucosidase [AG], two hydrolyze N-rich substrates N-acetyl-β-glucosaminidase [NAG] and leucine aminopeptidase [LAP]), and one hydrolyzes a P-rich substrate (acid phosphatase [PHOS]). Results showed decreased activities with depth for BG, CB, and LAP. Significantly higher potential enzyme activity was observed for convergent sites relative to planar or divergent sites across all depths sampled. Additionally, we looked at shifts in enzyme nutrient acquisition ratios that correspond with resource limitations relative to microbial stoichiometric demands. Higher acquisition potential is interpreted as greater resource allocation towards nutrient acquisition. Results showed a variance in resource acquisition potential with depth for C relative to N, with greater resources being allocated towards acquiring C at shallower depth. Conversely

  11. Impact of nitrogen pollution/deposition on extracellular enzyme activity, microbial abundance and carbon storage in coastal mangrove sediment.

    Science.gov (United States)

    Luo, Ling; Meng, Han; Wu, Ruo-Nan; Gu, Ji-Dong

    2017-06-01

    This study applied different concentration of NaNO3 solution to simulate the effect of inorganic nitrogen (N) deposition/pollution on carbon (C) storage in coastal mangrove sediment through observing the changes of enzyme activity and microbial abundance. Sediment collected from mangrove forest (MG) and intertidal zone (IZ) were incubated with different N rates (0 (control), 5 (low-N) and 20 (high-N) μg N g(-1) dry sediment, respectively). After incubation, the activities of phenol oxidase (PHO) and acid phosphatase (ACP) were enhanced, but β-glucosidase (GLU) and N-β-acetyl-glucosaminidase (NAG) activities were reduced by N addition. The altered enzymatic stoichiometries by N input implied that microbial phosphorus (P) limitation was increased, whereas C and N limitation were alleviated. Besides, N input decreased the bacterial abundance but increased fungal abundance in both types of sediment. The increased pH and soluble phenolics along with the exacerbated P limitation by N addition might explain these changes. Furthermore, sediment with N addition (except high-N treated MG sediment) showed a trend of C sequestration, which might be largely caused by the decrease of bacterial abundance and GLU activity. However, MG sediment with high-N suggested a trend of C loss, and the possible reason for this discrepancy might be the relatively higher increase of PHO and ACP activity. To better understand the influence of N deposition/pollution on C cycling, the long-term N effect on microorganisms, enzymes, and thus C storage should be paid more attention in the future. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Disruption of microbial biofilms by an extracellular protein isolated from epibiotic tropical marine strain of Bacillus licheniformis.

    Directory of Open Access Journals (Sweden)

    Devendra H Dusane

    Full Text Available BACKGROUND: Marine epibiotic bacteria produce bioactive compounds effective against microbial biofilms. The study examines antibiofilm ability of a protein obtained from a tropical marine strain of Bacillus licheniformis D1. METHODOLOGY/PRINCIPAL FINDINGS: B. licheniformis strain D1 isolated from the surface of green mussel, Perna viridis showed antimicrobial activity against pathogenic Candida albicans BH, Pseudomonas aeruginosa PAO1 and biofouling Bacillus pumilus TiO1 cultures. The antimicrobial activity was lost after treatment with trypsin and proteinase K. The protein was purified by ultrafiltration and size-exclusion chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF analysis revealed the antimicrobial agent to be a 14 kDa protein designated as BL-DZ1. The protein was stable at 75°C for 30 min and over a pH range of 3.0 to 11.0. The sequence alignment of the MALDI-fingerprint showed homology with the NCBI entry for a hypothetical protein (BL00275 derived from B. licheniformis ATCC 14580 with the accession number gi52082584. The protein showed minimum inhibitory concentration (MIC value of 1.6 µg/ml against C. albicans. Against both P. aeruginosa and B. pumilus the MIC was 3.12 µg/ml. The protein inhibited microbial growth, decreased biofilm formation and dispersed pre-formed biofilms of the representative cultures in polystyrene microtiter plates and on glass surfaces. CONCLUSION/SIGNIFICANCE: We isolated a protein from a tropical marine strain of B. licheniformis, assigned a function to the hypothetical protein entry in the NCBI database and described its application as a potential antibiofilm agent.

  13. Disruption of Microbial Biofilms by an Extracellular Protein Isolated from Epibiotic Tropical Marine Strain of Bacillus licheniformis

    Science.gov (United States)

    Dusane, Devendra H.; Damare, Samir R.; Nancharaiah, Yarlagadda V.; Ramaiah, N.; Venugopalan, Vayalam P.; Kumar, Ameeta Ravi; Zinjarde, Smita S.

    2013-01-01

    Background Marine epibiotic bacteria produce bioactive compounds effective against microbial biofilms. The study examines antibiofilm ability of a protein obtained from a tropical marine strain of Bacillus licheniformis D1. Methodology/Principal Findings B. licheniformis strain D1 isolated from the surface of green mussel, Perna viridis showed antimicrobial activity against pathogenic Candida albicans BH, Pseudomonas aeruginosa PAO1 and biofouling Bacillus pumilus TiO1 cultures. The antimicrobial activity was lost after treatment with trypsin and proteinase K. The protein was purified by ultrafiltration and size-exclusion chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis revealed the antimicrobial agent to be a 14 kDa protein designated as BL-DZ1. The protein was stable at 75°C for 30 min and over a pH range of 3.0 to 11.0. The sequence alignment of the MALDI-fingerprint showed homology with the NCBI entry for a hypothetical protein (BL00275) derived from B. licheniformis ATCC 14580 with the accession number gi52082584. The protein showed minimum inhibitory concentration (MIC) value of 1.6 µg/ml against C. albicans. Against both P. aeruginosa and B. pumilus the MIC was 3.12 µg/ml. The protein inhibited microbial growth, decreased biofilm formation and dispersed pre-formed biofilms of the representative cultures in polystyrene microtiter plates and on glass surfaces. Conclusion/Significance We isolated a protein from a tropical marine strain of B. licheniformis, assigned a function to the hypothetical protein entry in the NCBI database and described its application as a potential antibiofilm agent. PMID:23691235

  14. Indication of temperature inverted microbial assimilative capacities (extracellular enzymes activities in the pelagic of Lake Sevan (Armenia

    Directory of Open Access Journals (Sweden)

    Arevik MINASYAN

    2016-06-01

    Full Text Available Pioneering records of extracellular enzymes activities (EEA in Lake Sevan waters highlight dependence of heterotrophic functioning on physicochemical characteristics and bacterial assemblage. Values of EEA, ranged 0.11-30.39 µg C/P L-1h-1, were higher in upper layers compared to the omission in deeper parts. Particles associated (ecto- enzymes mainly predominated over free dissolved (exo- enzymes. In June activities of all studied enzymes followed similar pattern, particularly, decreasing at thermocline and increasing twice/more in cold deeper waters. Regardless higher bacterial density and temperature in June, with no similar records up to now, EEA revealed reverse relationship to temperature and bacteria data and were significantly lesser than in March. Our finding might be suggested as temperature inverted impact to heterotrophic activities in eutrophic conditions. We assume that observed, with temperature raise, declined EEA was due to blocked enzymatic active center from colloids and DOM components interaction, which, in overall, may suppress organic substrate utilization and result in weakening of first and rate limiting step of biological self-purification in Lake Sevan waters. Therefore, since temperature is co-regulator of assimilative/carrying capacity of aquatic ecosystems, climate warming might have unexpected negative feedbacks also through lowering assimilative capacities of water bodies, jeopardizing their quality and ecology.

  15. Polysaccharides enriched in rare sugars: bacterial sources, production and applications

    Directory of Open Access Journals (Sweden)

    Christophe eRoca

    2015-04-01

    Full Text Available Microbial extracellular polysaccharides (EPS, produced by a wide range of bacteria, are high molecular weight biopolymers, presenting an extreme diversity in terms of chemical structure and composition. They may be used in many applications, depending on their chemical and physical properties. A rather unexplored aspect is the presence of rare sugars in the composition of some EPS. Rare sugars, such as rhamnose or fucose, may provide EPS with additional biological properties compared to those composed of more common sugar monomers.This review gives a brief overview of these specific EPS and their producing bacteria. Cultivation conditions are summarized, demonstrating their impact on the EPS composition, together with downstream processing. Finally, their use in different areas, including cosmetics, food products, pharmaceuticals and biomedical applications, are discussed.

  16. Response of microbial extracellular enzyme activities and r- vs. K- selected microorganisms to elevated atmospheric CO2 depends on soil aggregate size

    Science.gov (United States)

    Dorodnikov, Maxim; Blagodatskaya, Evgenia; Blagodatskiy, Sergey; Kuzyakov, Yakov

    2014-05-01

    Increased belowground carbon (C) transfer by plant roots under elevated atmospheric CO2 and the contrasting environment in soil macro- and microaggregates could affect properties of the microbial community in the rhizosphere. We evaluated the effect of 5 years of elevated CO2 (550 ppm) on four extracellular enzymes: ß-glucosidase, chitinase, phosphatase, and sulfatase along with the contribution of fast- (r-strategists) and slow-growing microorganisms (K-strategists) in soil aggregates. We fractionated the bulk soil from the ambient and elevated CO2 treatments of FACE-Hohenheim (Stuttgart) into large macro- (>2 mm), small macro- (0.25-2.00 mm), and microaggregates (soil and aggregates amended with glucose and nutrients. In the bulk soil and isolated aggregates before and after activation with glucose, the actual and the potential enzyme activities were measured. Although C-org and C-mic as well as the activities of ß-glucosidase, phosphatase, and sulfatase were unaffected in bulk soil and in aggregate-size classes by elevated CO2, significant changes were observed in potential enzyme production after substrate amendment. After adding glucose, enzyme activities under elevated CO2 were 1.2-1.9-fold higher than under ambient CO2. In addition, µ values were significantly higher under elevated than ambient CO2 for bulk soil, small macroaggregates, and microaggregates. Based on changes in µ, GMB, and lag-period, we conclude that elevated atmospheric CO2 stimulated the r-selected microorganisms, especially in soil microaggregates. In contrast, significantly higher chitinase activity in bulk soil and in large macroaggregates under elevated CO2 revealed an increased contribution of fungi to turnover processes. We conclude that quantitative and qualitative changes of C input by plants into the soil at elevated CO2 affect microbial community functioning, but not its total content. An increase in r-selected microorganisms could accelerate C turnover in terrestrial

  17. Tailoring hierarchically porous graphene architecture by carbon nanotube to accelerate extracellular electron transfer of anodic biofilm in microbial fuel cells

    Science.gov (United States)

    Zou, Long; Qiao, Yan; Wu, Xiao-Shuai; Li, Chang Ming

    2016-10-01

    To overcoming their respective shortcomings of graphene and carbon nanotube, a hierarchically porous multi-walled carbon nanotube@reduced graphene oxide (MWCNT@rGO) hybrid is fabricated through a versatile and scalable solvent method, in which the architecture is tailored by inserting MWCNTs as scaffolds into the rGO skeleton. An appropriate amount of inserted 1-D MWCNTs not only effectively prevent the aggregation of rGO sheets but also act as bridges to increase multidirectional connections between 2-D rGO sheets, resulting in a 3-D hierarchically porous structure with large surface area and excellent biocompatibility for rich bacterial biofilm and high electron transfer rate. The MWCNT@rGO1:2/biofilm anode delivers a maximum power density of 789 mW m-2 in Shewanella putrefaciens CN32 microbial fuel cells, which is much higher than that of individual MWCNT and rGO, in particular, 6-folder higher than that of conventional carbon cloth. The great enhancement is ascribed to a synergistic effect of the integrated biofilm and hierarchically porous structure of MWCNT@rGO1:2/biofilm anode, in which the biofilm provides a large amount of bacterial cells to raise the concentration of local electron shuttles for accelerating the direct electrochemistry on the 3-D hierarchically porous structured anodes.

  18. Extracellular polysaccharide production by Thraustochytrid protists

    Digital Repository Service at National Institute of Oceanography (India)

    Jain, R.; Raghukumar, S.; Tharanathan, R.; Bhosle, N.B.

    and thixotrophic paints), flocculants (in water clarification and ore extraction), foam stabilizers (in beer and fire-fighting fluids), gelling agents (in cell and enzyme technology and foods), hydrating agents (in cosmetics and pharmaceuticals), and as inhibitors...

  19. Application of extracellular lipopeptide biosurfactant produced by endophytic Bacillus subtilis K1 isolated from aerial roots of banyan (Ficus benghalensis) in microbially enhanced oil recovery (MEOR).

    Science.gov (United States)

    Pathak, Khyati V; Keharia, Hareshkumar

    2014-02-01

    Bacillus subtilis K1 isolated from aerial roots of banyan tree secreted mixture of surfactins, iturins and fengycins with high degree of heterogeneity. The extracellular extract consisting of mixture of these cyclic lipopeptides exhibited very good emulsification activity as well as excellent emulsion stability. The culture accumulated maximum surfactant up to 48 h of growth during batch fermentation in Luria broth. The emulsion of hexane, heptane and octane prepared using 48-h-old culture supernatant of B. subtilis K1 remained stable up to 2 days while emulsion of four stroke engine oil remained stable for more than a year. The critical micelle concentration of crude lipopeptide biosurfactant extracted by acid precipitation from 48-h-old fermentation broth of B. subtilis K1 was found to be 20.5 μg/mL. The biosurfactant activity was found to be stable at 100 °C for 2 h, over a pH range of 6-12 h and over an NaCl concentration up to 10 % (w/v). The application of biosurfactant on laboratory scale sand pack column saturated with four stroke engine oil resulted in ~43 % enhanced oil recovery, suggesting its suitability in microbially enhanced oil recovery.

  20. Extracellular enzyme kinetics scale with resource availability

    Science.gov (United States)

    Microbial community metabolism relies on external digestion, mediated by extracellular enzymes that break down complex organic matter into molecules small enough for cells to assimilate. We analyzed the kinetics of 40 extracellular enzymes that mediate the degradation and assimi...

  1. Oligo- and polysaccharide synthesis by Rhizobium leguminosarum and Rhizobium meliloti.

    OpenAIRE

    Breedveld, M W

    1992-01-01

    Rhizobium and Agrobacterium species are capable of synthesizing a variety of extracellular and cellular oligo- and polysaccharides. Changes in environmental conditions may all affect the composition, physical properties, and relative amounts of oligo- and polysaccharides. Interest in the field of Rhizobium polys accharides has resulted from a development in two distinct areas, (i) the role of oligo- and polysaccharides in the microbe- plant interaction, and (ii) studies on the physico- chemic...

  2. Oligo- and polysaccharide synthesis by Rhizobium leguminosarum and Rhizobium meliloti

    NARCIS (Netherlands)

    Breedveld, M.W.

    1992-01-01

    Rhizobium and Agrobacterium species are capable of synthesizing a variety of extracellular and cellular oligo- and polysaccharides. Changes in environmental conditions may all affect the composition, physical properties, and relative amounts of

  3. Oligo- and polysaccharide synthesis by Rhizobium leguminosarum and Rhizobium meliloti.

    NARCIS (Netherlands)

    Breedveld, M.W.

    1992-01-01

    Rhizobium and Agrobacterium species are capable of synthesizing a variety of extracellular and cellular oligo- and polysaccharides. Changes in environmental conditions may all affect the composition, physical properties, and relative amounts of oligo- and polysaccharides. Interest in the field of Rh

  4. Extracellular polymeric substances enhanced mass transfer of polycyclic aromatic hydrocarbons in the two-liquid-phase system for biodegradation.

    Science.gov (United States)

    Zhang, Yinping; Wang, Fang; Yang, Xinglun; Gu, Chenggang; Kengara, Fredrick Orori; Hong, Qing; Lv, Zhengyong; Jiang, Xin

    2011-05-01

    The objective was to elucidate the role of extracellular polymeric substances (EPS) in biodegradation of polycyclic aromatic hydrocarbons in two-liquid-phase system (TLPs). Therefore, biodegradation of phenanthrene (PHE) was conducted in a typical TLPs--silicone oil-water--with PHE-degrading bacteria capable of producing EPS, Sphingobium sp. PHE3 and Micrococcus sp. PHE9. The results showed that the presence of both strains enhanced mass transfer of PHE from silicone oil to water, and that biodegradation of PHE mainly occurred at the interfaces. The ratios of tightly bound (TB) proteins to TB polysaccharides kept almost constant, whereas the ratios of loosely bound (LB) proteins to LB polysaccharides increased during the biodegradation. Furthermore, polysaccharides led to increased PHE solubility in the bulk water, which resulted in an increased PHE mass transfer. Both LB-EPS and TB-EPS (proteins and polysaccharides) correlated with PHE mass transfer in silicone oil, indicating that both proteins and polysaccharides favored bacterial uptake of PHE at the interfaces. It could be concluded that EPS could facilitate microbial degradation of PHE in the TLPs.

  5. Microbial mannanases: an overview of production and applications.

    Science.gov (United States)

    Dhawan, Samriti; Kaur, Jagdeep

    2007-01-01

    Microbial mannanases have become biotechnologically important since they target the hydrolysis of complex polysaccharides of plant tissues into simple molecules like manno-oligosaccharides and mannoses. The role of mannanases in the paper and pulp industry is well established and recently they have found application in the food and feed technology, coffee extraction, oil drilling and detergent industry. Mannanses are enzymes produced mainly from microorganisms but mannanases produced from plants and animals have also been reported. Bacterial mannanases are mostly extracellular and can act in a wide range of pH and temperature, though acidic and neutral mannanases are more common. This review will focus on complex mannan structure and the microbial enzyme complex involved in its complete breakdown, mannanase sources, production conditions and their applications in the commercial sector. The reference to plant and animal mannanases has been made to complete the overview. However, the major emphasis of the review is on the microbial mannanases.

  6. Separation and Purification of Intracellular and Extracellular Polysaccharides from Isochrysis galbana and Their Antimicrobial Activity%球等鞭金藻胞内和胞外多糖的分离纯化及其抑菌活性

    Institute of Scientific and Technical Information of China (English)

    孙颖颖; 周豹; 徐深圳; 李文浩; 阎斌伦

    2012-01-01

    采用分级沉淀、Sephadex G-75凝胶柱层析和DEAE-52离子交换柱层析对球等鞭金藻胞内粗多糖(IPS)进行分离;对胞外粗多糖(IEPS)则进行Sephadex G-100凝胶柱层析分离。在此基础上,分析所获多糖组分对大肠杆菌、枯草芽孢杆菌、金黄色葡萄球菌和变形杆菌等细菌的抑菌活性。同时,通过紫外光谱和红外光谱测定,比较IPS和IEPS结构的异同。结果表明:IPS和IEPS均具有抑菌活性,前者抑制大肠杆菌、枯草芽孢杆菌和金黄色葡萄球菌的生长,后者抑制大肠杆菌和枯草芽孢杆菌的生长;IPS经DEAE-52纤维素柱层析分离,获得2个多糖组分:IPSⅠ和IPSⅡ。IEPS经Sephadex G-100凝胶柱层析分离,获得3个多糖组分:IEPSA、IEPSB和IEPSC。活性检测表明,IPSⅠ和IPSⅡ抑制枯草芽孢杆菌和金黄色葡萄球菌的生长;IEPSA、IEPSB和IEPSC抑制大肠杆菌的生长;紫外光谱表明,IPS和IEPS均不含核酸、游离或裸露的蛋白质和色素;IPS和IEPS的红外光谱比较相似,以半乳糖为构架,均含有酰氨取代基,但不同之处在于,后者不含硫酸取代基。%Two crude polysaccharides,intracellular polysaccharide(IPS) and extracellular polysaccharide(EPS),were isolated from the cells and culture supernatant of Isochrysis galbana through hot-water extraction.The separation and purification of IPS and EPS,and their antimicrobial activities against Escherichia coli,Bacillus subtilis,Staphylococcus aureus and Proteus sp.were investigate,respectively.IPS was separated by ethanol-precipitation and purified by Sephadex G-75 gel chromatography,or by DEAE-52 column chromatography,while EPS was fractionated by Sephadex G-100 gel filtration chromatography.Finally,IPS and EPS were analyzed by UV and IR spectroscopy.The results indicated that: 1) IPS could inhibit the growth of Escherichia coli,Bacillus subtilis and Staphylococcus aureus.EPS could inhibit the growth of Escherichia coli and

  7. Sulfated polysaccharides and immune response: promoter or inhibitor?

    Science.gov (United States)

    Chen, D; Wu, X Z; Wen, Z Y

    2008-06-01

    Sulfated polysaccharides, which frequently connect to core protein, are expressed not only on cell surface but also throughout the extracellular matrix. Besides providing structural integrity of cells, sulfated polysaccharides interact with a variety of sulfated polysaccharides-binding proteins, such as growth factors, cytokines, chemokines and proteases. Sulfated polysaccharides play two-edged roles, inhibitor and promoter, in immune response. Some sulfated polysaccharides act as the immunosuppressor by blocking inflammatory signal transduction induced by proinflammatory cytokines, suppressing the activation of complement and inhibiting the process that leukocytes adhere to and pass through endothelium. On the contrary, the interaction between immune cells and sulfated polysaccharides produced by bacteria, endothelial cells and immune cells initiate the occurrence of immune response. It promotes the processes of recognizing and arresting antigen, migrating transendothelium, moving into and out of immune organ and enhancing the proliferation of lymphocyte. The structure of sulfated polysaccharides, such as molecular weight and sulfated sites heterogeneity, especially the degree of disaccharide sulfation, position of the sulfate moiety and organization of sulfated domains, may play critical role in their controversial effects. As a consequence, the interaction between sulfated polysaccharides and sulfated polysaccharide-binding proteins may be changed by modifying the structure of sulfated polysaccharides chains. The administration of drug targeting sulfated polysaccharide-protein interaction may be useful in treating inflammatory related diseases.

  8. 海洋微藻裂壶藻发酵液胞外多糖的制备及活性研究%Preparation of extracellular polysaccharide from fermentation liquor of marine microalgae Schizochytrium and study on the bioactivities

    Institute of Scientific and Technical Information of China (English)

    汪少芸; 江勇; 孟春; 欧阳艳华; 林祥志

    2011-01-01

    使用乙醇沉淀法从海洋微藻裂壶藻Schizochytrium sp.TIO1101的发酵液中提取胞外多糖,再经732阳离子交换树脂分离得到酸性多糖、弱碱性多糖和碱性多糖三种主要的多糖组分,并分别测定其还原力活性、DPPH自由基清除活性和羟基自由基清除活性等抗氧化活性和抗菌活性.结果表明,弱碱性多糖具有最高的抗氧化活性.抑制真菌活性实验表明,三种多糖组分对苹果轮纹病菌(Physalospora piricola)、瓜果腐霉病菌(Pythium aphanitermatum)、葡萄灰霉病菌(Botrytis cinerea)和甜瓜枯萎病菌(Fusarium pathogens)都表现出抑菌活性,并且对葡萄灰霉病菌(Botrytis cinerea)的抑菌效果最为明显.%The ethanol precipitation was executed to extract exopolysaccharide from the fermentation solution of marine microalgae Schizochytrium sp. TI01101. The extracted exopolysaccharide solution was then applied to 732 cationic exchange resin for further separation, the elution was isolated with in to three major polysaccharide components; acidic polysaccharide, weak alkaline polysaccharides and alkaline polysaccharides. And the polysaccharides were assessed for antioxidant activity against reduc ing power activity, DPPH radical scavenging activity and hydroxyl radical scavenging activity assays, respectively. The results indicated that weak alkaline polysaccharides showed significant antioxidant activity. Antifungal activity experiment showed all three polysaccharide components demonstrated antifungal activity to Physalospora piricola, Pythium aphanidermatum, Botrytis cinerea and Fusarium pathogens to some extent, particularly showing high effect on Botrytis cinerea.

  9. Impact of fermentation and addition of non-starch polysaccharide-degrading enzymes on microbial population and on digestibility of dried distillers grains with solubles in pigs

    DEFF Research Database (Denmark)

    Venås Jakobsen, Grethe; Jensen, Bent Borg; Knudsen, Knud Erik Bach;

    2015-01-01

    or with the addition of carbohydrases) and 40% of a basal diet composed mainly of maize starch: (1) a nonfermented liquid feed treatment (n-FLF); (2) a fermented liquid feed treatment (FLF); (3) the FLF supplemented with a mixture of xylanase and β-glucanase (XylGlu); and (4) the FLF supplemented with a mixture......-starch polysaccharides (NSP) compared to the fermented DDGS (from 341.8 g/kg DM to 312.9 g/kg DM, PFermenting DDGS prior to feeding led to significantly......Fluctuating prices on feedstock has led to a growing interest in alternative feed ingredients. Co-products from the biofuel industry are hence interesting to include in pig feeds, primarily due to the high protein content. Low nutritional value due to a high content of dietary fibre, however...

  10. Microbial control over carbon cycling in soil

    Directory of Open Access Journals (Sweden)

    Joshua eSchimel

    2012-09-01

    Full Text Available A major thrust of terrestrial microbial ecology is focused on understanding when and how the composition of the microbial community affects the functioning of biogeochemical processes at the ecosystem scale (meters-to-kilometers and days-to-years. While research has demonstrated these linkages for physiologically and phylogenetically narrow processes such as trace gas emissions and nitrification, there is less conclusive evidence that microbial community composition influences the broad processes of decomposition and organic matter turnover in soil. In this paper, we consider how soil microbial community structure influences C-cycling. We consider the phylogenetic level at which microbes form meaningful guilds, based on overall life history strategies, and suggest that these are associated with deep evolutionary divergences, while much of the species-level diversity probably reflects functional redundancy. We then consider under what conditions it is possible for differences among microbes to affect process dynamics, and argue that while microbial community structure may be important in the rate of OM breakdown in the rhizosphere and in detritus, it is likely not important in the mineral soil. In mineral soil, physical access to occluded or sorbed substrates is the rate-limiting process. Microbial community influences on OM turnover in mineral soils are based on how organisms allocate the C they take up—not only do the fates of the molecules differ, but they can affect the soil system differently as well. For example, extracellular enzymes and extracellular polysaccharides can be key controls on soil structure and function. How microbes allocate C may also be particularly important for understanding the long-term fate of C in soil—is it sequestered or not?

  11. Effects of Impellor Speed and Aeration Rate on Mycelial Biomass and Extracellular Polysaccharide Production,Reducing Sugar Consumption and Dissolved Oxygen Levels in Fermentor-grown Cultures of Phellinus baumii%不同搅拌转速和通气量对桑黄深层发酵培养的影响

    Institute of Scientific and Technical Information of China (English)

    雷萍; 吴亚召; 张文隽; 陈旭; 吕德平; 安军民

    2014-01-01

    The effects of impellor speed and aeration rate on mycelial biomass and extracellular polysaccharide production,reducing sugar consumption and dissolved oxygen levels during growth of Phellinus baumii in a 20-L fermentor were determined.Highest biomass (12.65 g/L)and extracellular polysaccharide(2.99 g/L) yields,and the most rapid decrease in dissolved oxygen levels,were recorded when the impellor speed was set at 150 r/min.Mycelial pellets were globose and compact,and the proportion of hyphal filaments was low. Highest yields of mycelial biomass (12.69 g/L)and extracellular polysaccharide(3.0 g/L),and the most rapid decrease in dissolved oxygen levels,were recorded when the aeration rate was set at 1∶0.65 vvm (air volume/culture volume/min).At this setting,the fungal mycelium grew well and formed compact pellets of uniform size.%探讨在20 L搅拌式发酵罐中,转速和通气量对桑黄菌丝生物量、胞外多糖产率、溶氧以及菌丝形态的影响。结果表明,实验范围内,转速为150 r/min时,桑黄菌丝体生物量最大(12.65 g/L),胞外多糖得率最高(2.99 g/L),相对溶氧下降最快,菌球小球状、较紧密、丝状体比例小;通气量为1∶0.65 vvm,桑黄菌丝体生物量最大(12.69 g/L),胞外多糖得率最高(3.00 g/L),相对溶氧下降最快,菌球大小均匀、紧密、生长良好。

  12. Extracellular proteins in pea root tip and border cell exudates.

    Science.gov (United States)

    Wen, Fushi; VanEtten, Hans D; Tsaprailis, George; Hawes, Martha C

    2007-02-01

    Newly generated plant tissue is inherently sensitive to infection. Yet, when pea (Pisum sativum) roots are inoculated with the pea pathogen, Nectria haematococca, most newly generated root tips remain uninfected even though most roots develop lesions just behind the tip in the region of elongation. The resistance mechanism is unknown but is correlated spatially with the presence of border cells on the cap periphery. Previously, an array of >100 extracellular proteins was found to be released while border cell separation proceeds. Here we report that protein secretion from pea root caps is induced in correlation with border cell separation. When this root cap secretome was proteolytically degraded during inoculation of pea roots with N. haematococca, the percentage of infected root tips increased from 4% +/- 3% to 100%. In control experiments, protease treatment of conidia or roots had no effect on growth and development of the fungus or the plant. A complex of >100 extracellular proteins was confirmed, by multidimensional protein identification technology, to comprise the root cap secretome. In addition to defense-related and signaling enzymes known to be present in the plant apoplast were ribosomal proteins, 14-3-3 proteins, and others typically associated with intracellular localization but recently shown to be extracellular components of microbial biofilms. We conclude that the root cap, long known to release a high molecular weight polysaccharide mucilage and thousands of living cells into the incipient rhizosphere, also secretes a complex mixture of proteins that appear to function in protection of the root tip from infection.

  13. Nonstarch polysaccharide-degrading enzymes alter the microbial community and the fermentation patterns of barley cultivars and wheat products in an in vitro model of the porcine gastrointestinal tract.

    Science.gov (United States)

    Bindelle, Jérôme; Pieper, Robert; Montoya, Carlos A; Van Kessel, Andrew G; Leterme, Pascal

    2011-06-01

    An in vitro experiment was carried out to assess how nonstarch polysaccharide (NSP)-degrading enzymes influence the fermentation of dietary fiber in the pig large intestine. Seven wheat and barley products and cultivars with differing carbohydrate fractions were hydrolyzed using pepsin and pancreatin in the presence or not of NSP-degrading enzymes (xylanase and β-glucanase) and the filter retentate was subsequently fermented with sow fecal bacteria. Dry matter, starch, crude protein and β-glucan digestibilities during hydrolysis were measured. Fermentation kinetics of the hydrolyzed ingredients were modelled. Short-chain fatty acids (SCFA) production and molar ratio were compared after 12, 24 and 72 h. Microbial communities were analyzed after 72 h of fermentation using terminal restriction fragment length polymorphism. The results showed an increase of nutrient digestibility (Penzyme. SCFA and bacterial community profiles also indicated a shift from propionate to acetate and an increase in cellulolytic Ruminococcus- and xylanolytic Clostridium-like bacteria. This is explained by the increase in slowly fermentable insoluble carbohydrate and the lower proportion of rapidly fermentable β-glucan and starch in the retentate when grains were incubated with NSP-degrading enzymes. Shifts were also different for the four barley varieties investigated, showing that the efficiency of the enzymes depends on the structure of the carbohydrate fractions in cereal products and cultivars.

  14. Importance of extracellular proteins in maintaining structural integrity of aerobic granules.

    Science.gov (United States)

    Xiong, Yanghui; Liu, Yu

    2013-12-01

    Aerobic granules developed through self-immobilization of microorganisms are compact and structured microbial consortia embedded in a matrix of extracellular polymeric substances (EPS). This study investigated the contribution of extracellular proteins (PN) to maintaining the structural integrity of aerobic granule. It was found that hydrolysis of PN induced by Proteinase K led to significant disintegration of aerobic granules, whereas a substantial reduction of extracellular polysaccharides (PS) was also observed. It was proposed that hydrolysis of extracellular proteins present in the EPS matrix of aerobic granules led to collapse of the EPS matrix, and subsequent disintegration of aerobic granule. These suggested that extracellular proteins would be essential for maintaining structural stability of EPS matrix of aerobic granules. In addition, it was revealed that production of signaling molecules, such as autoinducer-2 (AI-2) and N-acyl homoserine lactones (AHLs) was also inhibited probably due to hydrolysis of quorum sensing receptor proteins by Proteinase K. This in turn provided an additional explanation for the observed Proteinase K-triggered dispersal of aerobic granules.

  15. Belowground carbon allocation by trees drives seasonal patterns of extracellular enzyme activities by altering microbial community composition in a beech forest soil.

    Science.gov (United States)

    Kaiser, Christina; Koranda, Marianne; Kitzler, Barbara; Fuchslueger, Lucia; Schnecker, Jörg; Schweiger, Peter; Rasche, Frank; Zechmeister-Boltenstern, Sophie; Sessitsch, Angela; Richter, Andreas

    2010-08-01

    *Plant seasonal cycles alter carbon (C) and nitrogen (N) availability for soil microbes, which may affect microbial community composition and thus feed back on microbial decomposition of soil organic material and plant N availability. The temporal dynamics of these plant-soil interactions are, however, unclear. *Here, we experimentally manipulated the C and N availability in a beech forest through N fertilization or tree girdling and conducted a detailed analysis of the seasonal pattern of microbial community composition and decomposition processes over 2 yr. *We found a strong relationship between microbial community composition and enzyme activities over the seasonal course. Phenoloxidase and peroxidase activities were highest during late summer, whereas cellulase and protease peaked in late autumn. Girdling, and thus loss of mycorrhiza, resulted in an increase in soil organic matter-degrading enzymes and a decrease in cellulase and protease activity. *Temporal changes in enzyme activities suggest a switch of the main substrate for decomposition between summer (soil organic matter) and autumn (plant litter). Our results indicate that ectomycorrhizal fungi are possibly involved in autumn cellulase and protease activity. Our study shows that, through belowground C allocation, trees significantly alter soil microbial communities, which may affect seasonal patterns of decomposition processes.

  16. Marine Polysaccharide Networks and Diatoms at the Nanometric Scale

    Directory of Open Access Journals (Sweden)

    Tea Mišić Radić

    2013-10-01

    Full Text Available Despite many advances in research on photosynthetic carbon fixation in marine diatoms, the biophysical and biochemical mechanisms of extracellular polysaccharide production remain significant challenges to be resolved at the molecular scale in order to proceed toward an understanding of their functions at the cellular level, as well as their interactions and fate in the ocean. This review covers studies of diatom extracellular polysaccharides using atomic force microscopy (AFM imaging and the quantification of physical forces. Following a brief summary of the basic principle of the AFM experiment and the first AFM studies of diatom extracellular polymeric substance (EPS, we focus on the detection of supramolecular structures in polysaccharide systems produced by marine diatoms. Extracellular polysaccharide fibrils, attached to the diatom cell wall or released into the surrounding seawater, form distinct supramolecular assemblies best described as gel networks. AFM makes characterization of the diatom polysaccharide networks at the micro and nanometric scales and a clear distinction between the self-assembly and self-organization of these complex systems in marine environments possible.

  17. 球等鞭金藻胞外多糖的体外抗氧化活性和理化性质的初步分析%Study of in vitro antioxidation and physical and chemical characteristics analysis of extracellular polysaccharides iso-lated from Isochrysis galbana

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

    The extracellular polysaccharides, ECPSⅢ, has been successfully isolated from marine microalgae Isochrysis galbana through a combination of anion-exchange column chromatography and repeated gel chromatog-raphy in previous study. Its in vitro antioxidant activities and physical and chemical properties were investigated. The results showed that ECPSⅢ has good antioxidant activities and perticularly it has relative stronger scavenging activities against superoxide radical and hydroxyl radical. ECPSⅢ is a acidic polysaccharide rich in sulfuric acid radicals and uronic acid. The contents of sulfuric acid radicals and uronic acid were 76.90 mg/g and 17.1%in ECPSⅢ, respectively.%  前期研究采用离子交换柱层析和凝胶柱层析,制备到一种球等鞭金藻胞外纯多糖ECPSⅢ。在此基础上,采用化学比色法研究了 ECPSⅢ的体外抗氧化活性;同时,通过测定 ECPSⅢ中的硫酸基和糖醛酸含量,初步分析了ECPSⅢ的理化性质。结果表明, ECPSⅢ具有清除超氧阴离子(O2·¯)、羟基自由基(·OH)和过氧化氢(H2O2)等活性氧的能力和一定的还原力。其中, ECPSⅢ对O2·¯和·OH的清除能力较强。ECPSⅢ中的硫酸基和糖醛酸含量分别为76.90 mg/g和17.1%,是一种富含硫酸基和糖醛酸的酸性多糖。

  18. Application of fluorescently labelled lectins for the study of polysaccharides in biofilms with a focus on biofouling of nanofiltration membranes

    Directory of Open Access Journals (Sweden)

    Patrick Di Martino

    2016-07-01

    Full Text Available The biofilm state is the dominant microbial lifestyle in nature. A biofilm can be defined as cells organised as microcolonies embedded in an organic polymer matrix of microbial origin living at an interface between two different liquids, air and liquid, or solid and liquid. The biofilm matrix is made of extracellular polymeric substances, polysaccharides being considered as the major structural components of the matrix. Fluorescently labelled lectins have been widely used to stain microbial extracellular glycoconjugates in natural and artificial environments, and to study specific bacterial species or highly complex environments. Biofilm development at the membrane surface conducting to biofouling is one of the major problems encountered during drinking water production by filtration. Biofouling affects the durability and effectiveness of filtration membranes. Biofouling can be reduced by pretreatments in order to control two key parameters of water, the bioavailable organic matter concentration and the concentration of live bacteria. Nanofiltration (NF is a high technology process particularly suited to the treatment of surface waters to produce drinking water that is highly sensitive to biofouling. The development of strategies for fouling prevention and control requires characterizing the fouling material composition and organisation before and after NF membrane cleaning. The aim of this review is to present basics of biofilm analyses after staining with fluorescently labelled lectins and to focus on the use of fluorescent lectins and confocal laser scanning microscopy to analyse NF membrane biofouling.

  19. Seasonal Variation in Soil Microbial Biomass, Bacterial Community Composition and Extracellular Enzyme Activity in Relation to Soil Respiration in a Northern Great Plains Grassland

    Science.gov (United States)

    Wilton, E.; Flanagan, L. B.

    2014-12-01

    Soil respiration rate is affected by seasonal changes in temperature and moisture, but is this a direct effect on soil metabolism or an indirect effect caused by changes in microbial biomass, bacterial community composition and substrate availability? In order to address this question, we compared continuous measurements of soil and plant CO2 exchange made with an automatic chamber system to analyses conducted on replicate soil samples collected on four dates during June-August. Microbial biomass was estimated from substrate-induced respiration rate, bacterial community composition was determined by 16S rRNA amplicon pyrosequencing, and β-1,4-N-acetylglucosaminidase (NAGase) and phenol oxidase enzyme activities were assayed fluorometrically or by absorbance measurements, respectively. Soil microbial biomass declined from June to August in strong correlation with a progressive decline in soil moisture during this time period. Soil bacterial species richness and alpha diversity showed no significant seasonal change. However, bacterial community composition showed a progressive shift over time as measured by Bray-Curtis dissimilarity. In particular, the change in community composition was associated with increasing relative abundance in the alpha and delta classes, and declining abundance of the beta and gamma classes of the Proteobacteria phylum during June-August. NAGase showed a progressive seasonal decline in potential activity that was correlated with microbial biomass and seasonal changes in soil moisture. In contrast, phenol oxidase showed highest potential activity in mid-July near the time of peak soil respiration and ecosystem photosynthesis, which may represent a time of high input of carbon exudates into the soil from plant roots. This input of exudates may stimulate the activity of phenol oxidase, a lignolytic enzyme involved in the breakdown of soil organic matter. These analyses indicated that seasonal change in soil respiration is a complex

  20. POLYPEPTIDE AND POLYSACCHARIDE PROCESSING IN HYPERTHERMOPHILIC MICROORGANISMS

    Energy Technology Data Exchange (ETDEWEB)

    KELLY, ROBERT M.

    2008-12-22

    This project focused on the microbial physiology and biochemistry of heterotrophic hyperthermophiles with respect to mechanisms by which these organisms process polypeptides and polysaccharides under normal and stressed conditions. Emphasis is on two model organisms, for which completed genome sequences are available: Pyrococcus furiosus (growth Topt of 98°C), an archaeon, and Thermotoga maritima (growth Topt of 80°C), a bacterium. Both organisms are obligately anaerobic heterotrophs that reduce sulfur facultatively. Whole genome cDNA spotted microarrays were used to follow transcriptional response to a variety of environmental conditions in order to identify genes encoding proteins involved in the acquisition, synthesis, processing and utilization of polypeptides and polysaccharides. This project provided new insights into the physiological aspects of hyperthermophiles as these relate to microbial biochemistry and biological function in high temperature habitats. The capacity of these microorganisms to produce biohydrogen from renewable feedstocks makes them important for future efforts to develop biofuels.

  1. Isolation, Purification of Extracellular Polysaccharide from Viili and Its Effect on the Lifespan of Caenorhabditis elegans%Viili中胞外多糖的分离纯化及对秀丽线虫寿命的影响

    Institute of Scientific and Technical Information of China (English)

    王昌禄; 韩晓梅; 陈勉华; 王玉荣; 李风娟; 韩洪杰; 罗成

    2011-01-01

    The extracelluar polysaccharide (EPS) from Viili was obtained by Sevage method, ethanol deposition, the DEAE gel chromatography column and Sephadex G - 200 chromatography column. Then the effect of Viili EPS on longevity of Caenorhabditis elegans was detected by lifespan test. At 20℃, the mean lifespan of C. elegans was 17.13 ±0.53 d with Viili EPS 200 μg · mL-1. Compared with the control, the Viili EPS- treated groups showed significant increase in the mean lifespan of C. elegans( p < O. 001 ).%采用Sevage法脱蛋白、乙醇沉淀、DEAE-纤维素离子交换柱层析和Sephadex G-200分子筛层析法,从Viili中分离纯化得到Viili中乳酸菌胞外多糖(EPS).采用秀丽线虫进行寿命实验,观察Viili EPS延长秀丽线虫寿命的效果,在20℃下,饲喂Viili EPS 200 mg·L-1组秀丽线虫的平均寿命为17.13±0.53 d,与空白组相比,Viili EPS能够显著延长线虫的平均寿命(p< 0.001).

  2. Extraction, Characterization, and Molecular Weight Determination of Senna tora (L.) Seed Polysaccharide

    OpenAIRE

    Pawar, Harshal A.; K. G. Lalitha

    2015-01-01

    The objective of the present work was extraction of polysaccharide from Senna tora L. seed and its characterization as a pharmaceutical excipient. Polysaccharide extraction was based on mechanical separation of the endosperm of seeds of Senna tora, water dissolution, centrifugation, and precipitation with acetone. Standard procedures were used to study the viscosity, micromeritic properties, and microbial bioburden. Accelerated stability study was carried out on isolated polysaccharide for si...

  3. 高粘性微生物多糖发酵生产的关键性因素%The Key Factors in the High Viscosity Microbial Polysaccharide Fermentation Production

    Institute of Scientific and Technical Information of China (English)

    张永刚; 吉武科; 武琳; 严希海; 万丽萍

    2013-01-01

    本文以三种性质相似的高粘性微生物多糖黄原胶、结冷胶、韦兰胶为例,分析探讨了影响发酵生产过程的关键因素。关键制约因素为物质传递效率。提出四点有效措施:(1)诱变筛选加快菌株生长速率;(2)基因工程手段改造菌株提高菌株代谢产胶能力及高粘度体系内物质吸收速率;(3)增加菌株细胞膜通透性,提高物质传递效率;(4)通过一定的措施在发酵生产时有效降低体系粘度但不影响产物性质,经纯化提取后产品流变学性质不变。%In this paper , through three similar nature of high viscous microbial polysaccharide xanthan gum,gellan gum,welan gum,key factors have been analyzed and discussed in fermentation production process.The key constraint is mass transfer efficiency .Four measures were put forward:(1) Mutagenesis and screening accelerate the growth rate of strain .( 2 ) Genetic engineering tools transform strain to improve the metabolic producing ability and the material absorbed rate of the strain in high viscosity system .( 3 ) Increasing in membrane permeability of the strain cell can improve the efficiency of mass transfer.(4) By some certain measures effectively reducing the viscosity of the system but not affecting the properties of the product , the product rheological properties can be unchanged after extraction and purification in the production of the fermentation .

  4. Composition analysis of fractions of extracellular polymeric substances from an activated sludge culture and identification of dominant forces affecting microbial aggregation

    Science.gov (United States)

    Guo, Xuan; Wang, Xu; Liu, Junxin

    2016-06-01

    Extracellular polymeric substances (EPS) appear to play a critical role in the formation of bioaggregates, such as sludge flocs, in activated sludge processes. Here, we systematically investigated the composition and chemical structure of various EPS fractions excreted from an activated sludge culture using multi-analysis techniques to examine the ability of the sludge to aggregate. Chemical analysis was used with a three-dimensional excitation emission matrix and Fourier transform infrared spectroscopy, applying inter-particle forces theory. The combined findings revealed that hydrophobic groups, especially protein-related N-H, were present in a greater proportion in tightly bound EPS (TB-EPS). This result, which explained the specificity of TB-EPS in the chemical structure, was consistent with data indicating that TB-EPS contained a large amount of protein-like substances (86.7 mg/g of mixed liquor volatile suspended solids, 39.7% of the total EPS). Subsequently, a novel experimental procedure was developed to pinpoint key inter-particle forces in sludge aggregation. The result revealed that hydrogen bonds are the predominant triggers that promote sludge aggregation. This comprehensive analysis indicated that hydrophobic proteins in TB-EPS are responsible for the critical role played by hydrogen bonds in sludge formation. Our findings highlight the need to elucidate the mechanisms of TB-EPS-mediated flocculation in future efforts.

  5. Bioproduction, characterization, anticancer and antioxidant activities of extracellular melanin pigment produced by newly isolated microbial cell factories Streptomyces glaucescens NEAE-H

    Science.gov (United States)

    El-Naggar, Noura El-Ahmady; El-Ewasy, Sara M.

    2017-01-01

    In this present study, a newly isolated strain, Streptomyces sp. NEAE-H, capable of producing high amount of black extracellular melanin pigment on peptone-yeast extract iron agar and identified as Streptomyces glaucescens NEAE-H. Plackett–Burman statistical design was conducted for initial screening of 17 independent (assigned) variables for their significances on melanin pigment production by Streptomyces glaucescens NEAE-H. The most significant factors affecting melanin production are incubation period, protease-peptone and ferric ammonium citrate. The levels of these significant variables and their interaction effects were optimized by using face-centered central composite design. The maximum melanin production (31.650 μg/0.1 ml) and tyrosinase activity (6089.10 U/ml) were achieved in the central point runs under the conditions of incubation period (6 days), protease-peptone (5 g/L) and ferric ammonium citrate (0.5 g/L). Melanin pigment was recovered by acid-treatment. Higher absorption of the purified melanin pigment was observed in the UV region at 250 nm. It appeared to have defined small spheres by scanning electron microscopy imaging. The maximum melanin yield was 350 mg dry wt/L of production medium. In vitro anticancer activity of melanin pigment was assayed against skin cancer cell line using MTT assay. The IC50 value was 16.34 ± 1.31 μg/ml for melanin and 8.8 ± 0.5 μg/ml for standard 5-fluorouracil. PMID:28195138

  6. Activity of glycosidases from freshwater heterotrophic microorganisms on the degradation of extracellular polysaccharide produced by Anabaena spiroides (Cyanobacteria Atividade de glicosidases liberadas por microorganismos heterotróficos de água doce na degradação do polissacarídeo extracelular produzido por Anabaena spiroides (Cyanobacteria

    Directory of Open Access Journals (Sweden)

    Vanessa Colombo

    2004-06-01

    Full Text Available The activity of specific glycosidases during the degradation of the extracellular polysaccharide (EPS produced by Anabaena spiroides was determined using MUF-substrates (MUF-monosaccharides. Polysaccharide degradation was found to occur in a two-phase process. The first consisted of high enzymatic activity that consumed 41% of the EPS at a relatively high rate, while the second consumed the remaining polysaccharide (59% at a slower rate. A transition phase from the higher to the slower degradation rates was marked by a replacement of bacterial populations from coccoid to bacillus cells. During the degradation process, the bacterial biomass increased with the decrease of EPS, as revealed by bacterial cell counts. The enzymatic activity detected through the substrates MUF-alpha-D- and MUF-beta-D-glucoside was higher than that detected by other substrates tested. The remaining glycosides were MUF-alpha-L-rhamnopyranoside, MUF-beta-D-galactoside, MUF-alpha-D-mannopyranoside, MUF-beta-D-fucoside, MUF-beta-D-mannopyranoside, MUF-alpha-L-arabinopyranoside, and MUF-beta-L-fucoside. The fluorescence emitted by each MUF-substrate was proportional to the concentration of the corresponding monosaccharide in A. spiroides EPS. This demonstrates the susceptibility of EPS produced by A. spiroides to enzymatic attack by bacterial populations.A atividade de glicosidases durante a degradação do polissacarídeo extracelular (EPS produzido por Anabaena spiroides foi detectada e quantificada utilizando-se MUF-substratos (MUF-monossacarídeos. O consumo total do polissacarídeo efetuou-se em duas fases, uma primeira de alta atividade enzimática que rapidamente consumiu 41% do polissacarídeo e uma segunda, mais lenta, que consumiu o polissacarídeo restante (59%. A mudança de fase coincidiu com a sucessão de uma população de bactérias cocóides por outra de bacilos. A biomassa bacteriana, quantificada por contagens de células, aumentou com a degradação do

  7. Nonstarch polysaccharide-degrading enzymes alter the microbial community and the fermentation patterns of barley cultivars and wheat products in an in vitro model of the porcine gastrointestinal tract

    NARCIS (Netherlands)

    Bindelle, J.; Pieper, R.; Montoya, C.A.; Geurts van Kessel, A.H.M.; Leterme, P.

    2011-01-01

    An in vitro experiment was carried out to assess how nonstarch polysaccharide (NSP)-degrading enzymes influence the fermentation of dietary fiber in the pig large intestine. Seven wheat and barley products and cultivars with differing carbohydrate fractions were hydrolyzed using pepsin and pancreati

  8. Expression and the Functional Study of Fusion Proteins α-Amylase and Hemolysin- αas an Application in Biofilm Polysaccharide Degradation

    Directory of Open Access Journals (Sweden)

    Gede Yuda Sugiarta

    2016-12-01

    Full Text Available Biofilm is an aggregate of consortium bacteria that adhere to each other on a surface. It is usually protected by the exopolysaccharide layer. Various invasive medical procedures, such as catheterization, endotracheal tube installation, and contact lens utilization, are vulnerable to biofilm infection. The National Institute of Health (NIH estimates 65% of all microbial infections are caused by biofilm. Periplasmic α-amylase (MalS is an enzyme that hydrolyzes α-1, 4-glicosidic bond in glycogen, starch, and others related polysaccharides in periplasmic space. Another protein called hemolysin-α (HlyA is a secretion signal protein on C terminal of particular peptide in gram negative bacteria. We proposed a novel recombinant plasmid expressing α-amylase and hemolysin-αfusion in pSB1C3 which is cloned into E.colito enable α-amylase excretion to extracellular for degrading biofilm polysaccharides content, as in starch agar. Microtiter assay was performed to analyze the reduction percentage of biofilm by adding recombinant E.coli into media. This system is more effective in degrading biofilm from gram positive bacteria i.e.: Bacillus substilis(30.21% and Staphylococcus aureus (24.20%, and less effective degrading biofilm of gram negative i.e.: Vibrio cholera (5.30%, Pseudomonas aeruginosa (8.50%, Klebsiella pneumonia (6.75% andE. coli (-0.6%. Gram positive bacteria have a thick layer of peptidoglycan, causing the enzyme to work more effectively in degrading polysaccharides.

  9. Extracellular Electron Transfer from Aerobic Bacteria to Au-Loaded TiO2 Semiconductor without Light: A New Bacteria-Killing Mechanism Other than Localized Surface Plasmon Resonance or Microbial Fuel Cells.

    Science.gov (United States)

    Wang, Guomin; Feng, Hongqing; Gao, Ang; Hao, Qi; Jin, Weihong; Peng, Xiang; Li, Wan; Wu, Guosong; Chu, Paul K

    2016-09-21

    Titania loaded with noble metal nanoparticles exhibits enhanced photocatalytic killing of bacteria under light illumination due to the localized surface plasmon resonance (LSPR) property. It has been shown recently that loading with Au or Ag can also endow TiO2 with the antibacterial ability in the absence of light. In this work, the antibacterial mechanism of Au-loaded TiO2 nanotubes (Au@TiO2-NT) in the dark environment is studied, and a novel type of extracellular electron transfer (EET) between the bacteria and the surface of the materials is observed to cause bacteria death. Although the EET-induced bacteria current is similar to the LSPR-related photocurrent, the former takes place without light, and no reactive oxygen species (ROS) are produced during the process. The EET is also different from that commonly attributed to microbial fuel cells (MFC) because it is dominated mainly by the materials' surface, but not the bacteria, and the environment is aerobic. EET on the Au@TiO2-NT surface kills Staphylococcus aureus, but if it is combined with special MFC bacteria, the efficiency of MFC may be improved significantly.

  10. An Advance in the Research on Bioremediation of Heavy Metal Pollution by Microbial Extracellular Polymeric Substances%微生物胞外聚合物修复重金属污染研究进展

    Institute of Scientific and Technical Information of China (English)

    张广柱; 董鹏; 王繁业

    2009-01-01

    Extracellular polymeric substances (EPS) of mlcroblal origin are a complex mixture of biopoly-mers comprising polysaccharides, proteins, nucleic acids, uronic acids, humics, lipids, and etc. Functionally, EPS aid in cell-to-cell aggregation, adhesion to substratum, formation of floes, protection from desiccation and resistance to harmful exogenous materials. In addition, the exopolymers serve as bio-sorbing agents by accumulating nutrients from the surrounding environment and also play a crucial role in bio-sorption of heavy metals. Being poly-anionic in nature, EPS form complexes with metal cations resulting in metal immobilization. Moreover, enzymatic activities in EPS also assist detoxification of heavy metals by transformation and subsequent precipitation. Structures and functions of EPS were summarised focusing on their formation, separation and composition. Cationic bonding behaviour of EPS from various origins as well as the mechanism of remedying heavy metal pollution were presented.%微生物胞外聚合物(EPS)是由多糖、蛋白质、核酸、糖羧酸,腐殖质和脂类等生物聚合体组成的复杂混合物.EPS具有聚合细胞、固定基质、形成絮凝、保持水分及阻止有害外源物质等功能.此外,EPS还可作为生物吸附剂来吸附周围环境中营养物质,并且在重金属生物吸附中发挥关键作用.EPS为天然的聚阴离子物质.可固定金属离子形成复合物.EPS中的酶可通过转换和沉淀作用来加速重金属的去毒.文章综述了EPS的结构和功能,重点是其生成、分离、组成,介绍各种来源EPS的阳离子键合性能,及其重金属修复机理.

  11. The extracellular matrix of plants: Molecular, cellular and developmental biology

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1996-12-31

    A symposium entitled ``The Extracellular Matrix of Plants: Molecular, Cellular and Developmental Biology was held in Tamarron, Colorado, March 15--21, 1996. The following topics were explored in addresses by 43 speakers: structure and biochemistry of cell walls; biochemistry, molecular biology and biosynthesis of lignin; secretory pathway and synthesis of glycoproteins; biosynthesis of matrix polysaccharides, callose and cellulose; role of the extracellular matrix in plant growth and development; plant cell walls in symbiosis and pathogenesis.

  12. The Extracellular Matrix in Photosynthetic Mats: A Cyanobacterial Gingerbread House

    Science.gov (United States)

    Stuart, R.; Stannard, W.; Bebout, B.; Pett-Ridge, J.; Mayali, X.; Weber, P. K.; Lipton, M. S.; Lee, J.; Everroad, R. C.; Thelen, M.

    2014-12-01

    Hypersaline laminated cyanobacterial mats are excellent model systems for investigating photoautotrophic contributions to biogeochemical cycling on a millimeter scale. These self-sustaining ecosystems are characterized by steep physiochemical gradients that fluctuate dramatically on hour timescales, providing a dynamic environment to study microbial response. However, elucidating the distribution of energy from light absorption into biomass requires a complete understanding of the various constituents of the mat. Extracellular polymeric substances (EPS), which can be composed of proteins, polysaccharides, lipids and DNA are a major component of these mats and may function in the redistribution of nutrients and metabolites within the community. To test this notion, we established a model mat-building culture for comparison with the phylogenetically diverse natural mat communities. In these two systems we determined how proteins and glycans in the matrix changed as a function of light and tracked nutrient flow from the matrix. Using mass spectrometry metaproteomics analysis, we found homologous proteins in both field and culture extracellular matrix that point to cyanobacterial turnover of amino acids, inorganic nutrients, carbohydrates and nucleic acids from the EPS. Other abundant functions identified included oxidative stress response from both the cyanobacteria and heterotrophs and cyanobacterial structural proteins that may play a role in mat cohesion. Several degradative enzymes also varied in abundance in the EPS in response to light availability, suggesting active secretion. To further test cyanobacterial EPS turnover, we generated isotopically-labeled EPS and used NanoSIMS to trace uptake of this labeled EPS. Our findings suggest Cyanobacteria may facilitate nutrient transfer to other groups, as well as uptake of their own products through degradation of EPS components. This work provides evidence for the essential roles of EPS for storage, structural

  13. Augmenting Plant Immune Responses and Biological Control by Microbial Determinants

    Directory of Open Access Journals (Sweden)

    Sang Moo Lee

    2015-09-01

    Full Text Available Plant have developed sophisticated defence mechanisms against microbial pathogens. The recent accumulated information allow us to understand the nature of plant immune responses followed by recognition of microbial factors/determinants through cutting-edge genomics and multi-omics techniques. However, the practical approaches to sustain plant health using enhancement of plant immunity is yet to be fully appreciated. Here, we overviewed the general concept and representative examples on the plant immunity. The fungal, bacterial, and viral determinants that was previously reported as the triggers of plant immune responses are introduced and described as the potential protocol of biological control. Specifically, the role of chitin, glucan, lipopolysaccharides/extracellular polysaccharides, microbe/pathogen-associated molecular pattern, antibiotics, mimic-phytohormones, N-acyl homoserine lactone, harpin, vitamins, and volatile organic compounds are considered. We hope that this review stimulates scientific community and farmers to broaden their knowledge on the microbial determinant-based biological control and to apply the technology on the integrated pest management program.

  14. Bioactivity and Applications of Sulphated Polysaccharides from Marine Microalgae

    Directory of Open Access Journals (Sweden)

    Rui Manuel Santos Costa de Morais

    2013-01-01

    Full Text Available Marine microalgae have been used for a long time as food for humans, such as Arthrospira (formerly, Spirulina, and for animals in aquaculture. The biomass of these microalgae and the compounds they produce have been shown to possess several biological applications with numerous health benefits. The present review puts up-to-date the research on the biological activities and applications of polysaccharides, active biocompounds synthesized by marine unicellular algae, which are, most of the times, released into the surrounding medium (exo- or extracellular polysaccharides, EPS. It goes through the most studied activities of sulphated polysaccharides (sPS or their derivatives, but also highlights lesser known applications as hypolipidaemic or hypoglycaemic, or as biolubricant agents and drag-reducers. Therefore, the great potentials of sPS from marine microalgae to be used as nutraceuticals, therapeutic agents, cosmetics, or in other areas, such as engineering, are approached in this review.

  15. Bioactivity and applications of sulphated polysaccharides from marine microalgae.

    Science.gov (United States)

    Raposo, Maria Filomena de Jesus; de Morais, Rui Manuel Santos Costa; Bernardo de Morais, Alcina Maria Miranda

    2013-01-23

    Marine microalgae have been used for a long time as food for humans, such as Arthrospira (formerly, Spirulina), and for animals in aquaculture. The biomass of these microalgae and the compounds they produce have been shown to possess several biological applications with numerous health benefits. The present review puts up-to-date the research on the biological activities and applications of polysaccharides, active biocompounds synthesized by marine unicellular algae, which are, most of the times, released into the surrounding medium (exo- or extracellular polysaccharides, EPS). It goes through the most studied activities of sulphated polysaccharides (sPS) or their derivatives, but also highlights lesser known applications as hypolipidaemic or hypoglycaemic, or as biolubricant agents and drag-reducers. Therefore, the great potentials of sPS from marine microalgae to be used as nutraceuticals, therapeutic agents, cosmetics, or in other areas, such as engineering, are approached in this review.

  16. Bioactivity and Applications of Sulphated Polysaccharides from Marine Microalgae

    Science.gov (United States)

    de Jesus Raposo, Maria Filomena; de Morais, Rui Manuel Santos Costa; de Morais, Alcina Maria Miranda Bernardo

    2013-01-01

    Marine microalgae have been used for a long time as food for humans, such as Arthrospira (formerly, Spirulina), and for animals in aquaculture. The biomass of these microalgae and the compounds they produce have been shown to possess several biological applications with numerous health benefits. The present review puts up-to-date the research on the biological activities and applications of polysaccharides, active biocompounds synthesized by marine unicellular algae, which are, most of the times, released into the surrounding medium (exo- or extracellular polysaccharides, EPS). It goes through the most studied activities of sulphated polysaccharides (sPS) or their derivatives, but also highlights lesser known applications as hypolipidaemic or hypoglycaemic, or as biolubricant agents and drag-reducers. Therefore, the great potentials of sPS from marine microalgae to be used as nutraceuticals, therapeutic agents, cosmetics, or in other areas, such as engineering, are approached in this review. PMID:23344113

  17. Export of Extracellular Polysaccharides Modulates Adherence of the Cyanobacterium Synechocystis

    Energy Technology Data Exchange (ETDEWEB)

    Fisher, ML; Allen, R; Luo, YQ; Curtiss, R

    2013-09-10

    The field of cyanobacterial biofuel production is advancing rapidly, yet we know little of the basic biology of these organisms outside of their photosynthetic pathways. We aimed to gain a greater understanding of how the cyanobacterium Synechocystis PCC 6803 (Synechocystis, hereafter) modulates its cell surface. Such understanding will allow for the creation of mutants that autoflocculate in a regulated way, thus avoiding energy intensive centrifugation in the creation of biofuels. We constructed mutant strains lacking genes predicted to function in carbohydrate transport or synthesis. Strains with gene deletions of slr0977 (predicted to encode a permease component of an ABC transporter), slr0982 (predicted to encode an ATP binding component of an ABC transporter) and slr1610 (predicted to encode a methyltransferase) demonstrated flocculent phenotypes and increased adherence to glass. Upon bioinformatic inspection, the gene products of slr0977, slr0982, and slr1610 appear to function in O-antigen (OAg) transport and synthesis. However, the analysis provided here demonstrated no differences between OAg purified from wild-type and mutants. However, exopolysaccharides (EPS) purified from mutants were altered in composition when compared to wild-type. Our data suggest that there are multiple means to modulate the cell surface of Synechocystis by disrupting different combinations of ABC transporters and/or glycosyl transferases. Further understanding of these mechanisms may allow for the development of industrially and ecologically useful strains of cyanobacteria. Additionally, these data imply that many cyanobacterial gene products may possess as-yet undiscovered functions, and are meritorious of further study.

  18. Study on the Optimum Fermentation Condition and Physico-Chemical Property of Extracellular Polysaccharide Producing Strain BIT-BP004%菌株BIT-BP004产胞外多糖影响因素及发酵液理化特性的研究

    Institute of Scientific and Technical Information of China (English)

    汪小平; 付娜; 王平; 赵静; 梅晓丹

    2012-01-01

    从辽河油田油水样品中分离得到一株高产胞外多糖的菌株BIT-BP004。通过对其培养条件的考察和优化。得到最适宜的培养条件为:蔗糖7.5g/L。NaNO30.3g/L,l(2HPO41.5g/L,KI-12PO43g/L,MgSO40.1g/L,CaCl20.01g/L,初始pH为6,接茵量1%。培养24h后的胞外多糖产量为4.11g/L。同时,通过正交实验考察了温度、矿化度和pH值对BIT-BP004发酵液理化性能的影响。结果表明,不同实验条件对发酵液黏度影响较大。影响能力大小依次为温度〉矿化度〉pH,其中温度是影响发酵液黏度的决定性因素。图5表5参10%An extracellular polysaccharide (EPS) producing bacteria, named BIT-BP004, was isolated from oil/water sample of Liaohe oilfield. The optimal medium compositions in shake scale for strain BIT-BP004 were determined as follows: sucrose 7.5 g/L, NaNO3 0.5 g/L, K2HPO4 1.5 g/L, KH2PO4 3 g/L, MgSO4 0.1 g/L, CaCl2 0.01 g/L, initial pH 6 and ineculant's volume 1%. The EPS yield was 4. 11 g/L in batch culture after 24 h. By orthogonal design, the factors of temperature, mineral salt concentration and pH value were selected to assay the physico-chemical property of fermentation. The results showed that experiment factors had a larger effect on viscosity of fermentation, and the affection capacity was arranged as follows, temperature 〉 mineral salt concentration 〉 pH, indicating that temperature was a decisive factor on the viscosity of the fermentation broth.

  19. Extracellular Electron Uptake: Among Autotrophs and Mediated by Surfaces

    DEFF Research Database (Denmark)

    Tremblay, Pier-Luc; Angenent, Largus T.; Zhang, Tian

    2017-01-01

    Autotrophic microbes can acquire electrons from solid donors such as steel, other microbial cells, or electrodes. Based on this feature, bioprocesses are being developed for the microbial electrosynthesis (MES) of useful products from the greenhouse gas CO2. Extracellular electron-transfer mechan......Autotrophic microbes can acquire electrons from solid donors such as steel, other microbial cells, or electrodes. Based on this feature, bioprocesses are being developed for the microbial electrosynthesis (MES) of useful products from the greenhouse gas CO2. Extracellular electron...

  20. CHANGES IN THE MORPHOLOGY AND POLYSACCHARIDE CONTENT OF MICROCYSTIS AERUGINOSA (CYANOBACTERIA) DURING FLAGELLATE GRAZING(1).

    Science.gov (United States)

    Yang, Zhou; Kong, Fanxiang; Shi, Xiaoli; Zhang, Min; Xing, Peng; Cao, Huansheng

    2008-06-01

    To investigate the changes in the morphology and polysaccharide content of Microcystis aeruginosa (Kütz.) Kütz. during flagellate grazing, cultures of M. aeruginosa were exposed to grazing Ochromonas sp. for a period of 9 d under controlled laboratory conditions. M. aeruginosa responded actively to flagellate grazing and formed colonies, most of which were made up of several or dozens of cells, suggesting that flagellate grazing may be one of the biotic factors responsible for colony formation in M. aeruginosa. When colonies were formed, the cell surface ultrastructure changed, and the polysaccharide layer on the surface of the cell wall became thicker. This change indicated that synthesis and secretion of extracellular polysaccharide (EPS) of M. aeruginosa cells increased under flagellate grazing pressure. The contents of soluble extracellular polysaccharide (sEPS), bound extracellular polysaccharide (bEPS), and total polysaccharide (TPS) in colonial cells of M. aeruginosa increased significantly compared with those in single cells. This finding suggested that the increased amount of EPS on the cell surface may play a role in keeping M. aeruginosa cells together to form colonies.

  1. Extracellular DNA metabolism in Haloferax volcanii

    Directory of Open Access Journals (Sweden)

    Scott eChimileski

    2014-02-01

    Full Text Available Extracellular DNA is found in all environments and is a dynamic component of the micro-bial ecosystem. Microbial cells produce and interact with extracellular DNA through many endogenous mechanisms. Extracellular DNA is processed and internalized for use as genetic information and as a major source of macronutrients, and plays several key roles within prokaryotic biofilms. Hypersaline sites contain some of the highest extracellular DNA con-centrations measured in nature–a potential rich source of carbon, nitrogen and phosphorus for halophilic microorganisms. We conducted DNA growth studies for the halophilic archaeon Haloferax volcanii DS2 and show that this model Halobacteriales strain is capable of using exogenous double-stranded DNA as a nutrient. Further experiments with varying medium composition, DNA concentration and DNA types revealed that DNA is utilized primarily as a phosphorus source, that growth on DNA is concentration-dependent and that DNA isolated from different sources is metabolized selectively, with a bias against highly divergent methylated DNA sources. Additionally, fluorescence microscopy experiments showed that labeled DNA colocalized with Haloferax volcanii cells. The gene Hvo_1477 was also identified using a comparative genomic approach as a factor likely to be involved in extracellular DNA processing at the cell surface, and deletion of Hvo_1477 created an H. volcanii strain deficient in its ability to grow on extracellular DNA. Widespread distribution of Hvo_1477 homologs in archaea suggests metabolism of extracellular DNA may be of broad ecological and physiological relevance in this domain of life.

  2. Characterization of cell surface and extracellular matrix remodeling of Azospirillum brasilense chemotaxis-like 1 signal transduction pathway mutants by atomic force microscopy.

    Science.gov (United States)

    Edwards, Amanda Nicole; Siuti, Piro; Bible, Amber N; Alexandre, Gladys; Retterer, Scott T; Doktycz, Mitchel J; Morrell-Falvey, Jennifer L

    2011-01-01

    To compete in complex microbial communities, bacteria must sense environmental changes and adjust cellular functions for optimal growth. Chemotaxis-like signal transduction pathways are implicated in the regulation of multiple behaviors in response to changes in the environment, including motility patterns, exopolysaccharide production, and cell-to-cell interactions. In Azospirillum brasilense, cell surface properties, including exopolysaccharide production, are thought to play a direct role in promoting flocculation. Recently, the Che1 chemotaxis-like pathway from A. brasilense was shown to modulate flocculation, suggesting an associated modulation of cell surface properties. Using atomic force microscopy, distinct changes in the surface morphology of flocculating A. brasilense Che1 mutant strains were detected. Whereas the wild-type strain produces a smooth mucosal extracellular matrix after 24 h, the flocculating Che1 mutant strains produce distinctive extracellular fibril structures. Further analyses using flocculation inhibition, lectin-binding assays, and comparison of lipopolysaccharides profiles suggest that the extracellular matrix differs between the cheA1 and the cheY1 mutants, despite an apparent similarity in the macroscopic floc structures. Collectively, these data indicate that disruption of the Che1 pathway is correlated with distinctive changes in the extracellular matrix, which likely result from changes in surface polysaccharides structure and/or composition.

  3. Characterization of cell surface and extracellular matrix remodeling of Azospirillum brasilense chemotaxis-like 1 signal transduction pathway mutants by atomic force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Doktycz, Mitchel John [ORNL; Morrell-Falvey, Jennifer L [ORNL

    2011-01-01

    To compete in complex microbial communities, bacteria must sense environmental changes and adjust cellular functions for optimal growth. Chemotaxis-like signal transduction pathways are implicated in the regulation of multiple behaviors in response to changes in the environment, including motility patterns, exopolysaccharide production, and cell-to-cell interactions. In Azospirillum brasilense, cell surface properties, including exopolysaccharide production, are thought to play a direct role in promoting flocculation. Recently, the Che1 chemotaxis-like pathway from A. brasilense was shown to modulate flocculation, suggesting an associated modulation of cell surface properties. Using atomic force microscopy, distinct changes in the surface morphology of flocculating A. brasilense Che1 mutant strains were detected. Whereas the wild-type strain produces a smooth mucosal extracellular matrix after 24 h, the flocculating Che1 mutant strains produce distinctive extracellular fibril structures. Further analyses using flocculation inhibition, lectin-binding assays, and comparison of lipopolysaccharides profiles suggest that the extracellular matrix differs between the cheA1 and the cheY1 mutants, despite an apparent similarity in the macroscopic floc structures. Collectively, these data indicate that disruption of the Che1 pathway is correlated with distinctive changes in the extracellular matrix, which likely result from changes in surface polysaccharides structure and/or composition.

  4. Sulfation patterns determine cellular internalization of heparin-like polysaccharides

    OpenAIRE

    Raman, Karthik; Mencio, Caitlin; Desai, Umesh R.; Kuberan, Balagurunathan

    2013-01-01

    Heparin is a highly sulfated polysaccharide which serves biologically relevant roles as an anticoagulant and anti-cancer agent. While it is well known that modification of heparin’s sulfation pattern can drastically influence its ability to bind growth factors and other extracellular molecules, very little is known about the cellular uptake of heparin and the role sulfation patterns serve in affecting its internalization. In this study, we chemically synthesized several fluorescently-labeled ...

  5. Effect of medium pH and cultivation period on mycelial biomass, polysaccharide, and ligninolytic enzyme production by Ganoderma lucidum from Montenegro

    Directory of Open Access Journals (Sweden)

    Vukojević Jelena

    2006-01-01

    Full Text Available The effect of initial medium pH on biomass, extracellular and intracellular polysaccharide, and ligninolytic enzyme production by Ganoderma lucidum was investigated at different pH values after 7 and 14 days of cultivation. Maximal production of biomass was recorded at pH 4.5 and 5.0; maximal production of extracellular polysaccharides at pH 7.0 and 3.0; and maximal production of intracellular polysaccharides at pH 7.0 and 5.5. Ligninolytic enzymes were not produced at any pH of the medium. Maximal biomass production was obtained on the 11th day of cultivation; maximal extracellular polysaccharide production on the 7th day; and maximal intracellular polysaccharide production on the 6th and 10th day of cultivation. .

  6. Polysaccharides from Probiotics: New Developments as Food Additives

    Directory of Open Access Journals (Sweden)

    Philippe Michaud

    2010-01-01

    Full Text Available Microbial polysaccharides with nutraceutical potential and bioactive properties have been investigated in detail during the last few decades. There is an increasing demand in food industries for live microbes or polysaccharides produced by them which assert health benefits other than dietetic constituents. Although there are a large number of exopolysaccharide (EPS-producing bacteria, the titers are low for commercialization. This manuscript deals with the polysaccharides produced by probiotic strains, with major emphasis on the EPSs, their properties, applications and some of the strategies adopted which would be helpful in better understanding of the process in the near future. Research on the improved EPS biosynthesis is essential for obtaining high yields. Therefore, to reach commercialization, metabolic engineering must be applied.

  7. Microbial ecology of phototrophic biofilms

    NARCIS (Netherlands)

    Roeselers, G.

    2007-01-01

    Biofilms are layered structures of microbial cells and an extracellular matrix of polymeric substances, associated with surfaces and interfaces. Biofilms trap nutrients for growth of the enclosed microbial community and help prevent detachment of cells from surfaces in flowing systems. Phototrophic

  8. Microbial ecology of phototrophic biofilms

    NARCIS (Netherlands)

    Roeselers, G.

    2007-01-01

    Biofilms are layered structures of microbial cells and an extracellular matrix of polymeric substances, associated with surfaces and interfaces. Biofilms trap nutrients for growth of the enclosed microbial community and help prevent detachment of cells from surfaces in flowing systems. Phototrophic

  9. Belowground response to drought in a tropical forest soil. II. Change in microbial function impacts carbon composition.

    Directory of Open Access Journals (Sweden)

    Nick eBouskill

    2016-03-01

    Full Text Available Climate model projections for tropical regions show clear perturbation of precipitation patterns leading to increased frequency and severity of drought in some regions. Previous work has shown declining soil moisture to be a strong driver of changes in microbial trait distribution, however the feedback of any shift in functional potential on ecosystem properties related to carbon cycling are poorly understood. Here we show that drought-induced changes in microbial functional diversity and activity shape, and are in turn shaped by, the composition of dissolved and soil-associated carbon. We also demonstrate that a shift in microbial functional traits that favor the production of hygroscopic compounds alter the efflux of carbon dioxide following soil rewetting. Under drought the composition of the dissolved organic carbon pool changed in a manner consistent with a microbial metabolic response. We hypothesize that this microbial ecophysiological response to changing soil moisture elevates the intracellular carbon demand stimulating extracellular enzyme production, that prompts the observed decline in more complex carbon compounds (e.g., cellulose and lignin. Furthermore, a metabolic response to drought appeared to condition (biologically and physically the soil, notably through the production of polysaccharides, particularly in experimental plots that had been pre-exposed to a short-term drought. This hysteretic response, in addition to an observed drought-related decline in phosphorus concentration, may have been responsible for a comparatively modest CO2 efflux following wet-up in drought plots relative to control plots.

  10. Neutrophils cast extracellular traps in response to protozoan parasites.

    Science.gov (United States)

    Abi Abdallah, Delbert S; Denkers, Eric Y

    2012-01-01

    Release of extracellular traps by neutrophils is a now well-established phenomenon that contributes to the innate response to extracellular bacterial and fungal pathogens. The importance of NETs during protozoan infection has been less explored, but recent findings suggest an emerging role for release of neutrophil-derived extracellular DNA in response to this class of microbial pathogens. The present review summarizes findings to date regarding elicitation of NETs by Toxoplasma gondii, Plasmodium falciparum, Eimeria bovis, and Leishmania spp.

  11. Antibacterial and antiviral study of dialdehyde polysaccharides

    Science.gov (United States)

    Song, Le

    Concerns for microbial contamination and infection to the general population, especially the spread of drug-resistant microorganisms, have greatly increased. Polymeric biocides have been found to be a feasible strategy to inactivate drug-resistant bacteria. However, current polymeric biocide systems involve multi-step chemical reactions and they are not cost-effective. Desirable antimicrobial systems need to be designed to be environmentally friendly, broad-spectrum effective against microorganisms, flexible for various delivery methods and economically affordable. We demonstrated that dialdehyde polysaccharides (including dialdehyde starch and dialdehdye cellulose) were broad-spectrum polymeric biocides against gram-positive/negative bacteria, bacteriophages and human virus. These polymers can be easily converted from starch and cellulose through one-step periodate oxidation. Destructions of microorganism by dialdehyde polysaccharides have been achieved in aqueous suspension or by solid surface contact. The dialdehdye functions of dialdehdye polysaccharides were found to be the dominant action against microorganism. The reactivity of the dialdehyde functionality was found to be pH-dependent as well as related to the dispersion of dialdehyde polysaccharides. Degradation of dialdehyde starch during cooking was confirmed. Degradation of dialdehyde starch was more liable in alkaline condition. Carboxylic acid and conjugated aldehyde functionalities were the two main degradation products, confirmed from the spectroscopic studies. The pH effect on the polysaccharide structure and the corresponding antimicrobial activity was very complicated. No decisive conclusions could be obtained from this study. Liner inactivation kinetics was found for dialdehyde starch aqueous suspension against bacteria. This linear inactivation kinetics was derived from the pseudo-first chemical reaction between the dialdehyde starch and the bacteria. The established inactivation kinetics was

  12. 铜绿假单胞菌 Psl 多糖转运蛋白 PslD 的表达与纯化研究%Expression and Purification of Extracellular Polysaccharides Psl Transporter PslD from Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    唐玮蔚; 马旅雁; 吴更

    2015-01-01

    The opportunistic Pseudomonas aeruginosa is a model organism for biofilm research. Its excreted extracel-lular polysaccharide Psl is a key component of biofilm matrix in P. aeruginosa. The Psl transporter PslD is composed of 256 amino acids,bio-informatics analysis has revealed that it comprised of one transmembrane protein having N ter-minal signal peptide. Detergents are required to isolate and purify transmembrane protein. However,detergents are various in kinds with different properties,in this study a set of scenario was designed for screening detergent to dis-solve Ps1D. Through anti-histidine labeled western blot analysis n-Decyl-β-D-maltopyranoside(DM),n-Decyl-N, Ndimethylamine-N-oxide( DDAO)and n-Dodecyl-N,N-dimethylamine-N-oxide( LDAO)were considered to have high extracting efficiency. By altering the ratio of total protein to the detergent,further optimal solubilization condition was determined as 8 mg/ mL total protein:1%(w/ v)LDAO. Under this soluble condition,over 80% purity can be achieved only through the first step of Ni column affinity chromatography to purify the goal protein,this had laid a foundation for study on its further crystallization attempt and its structural biology.%条件性致病菌铜绿假单胞菌是细菌生物被膜研究的模式菌,其分泌的胞外多糖 Psl 在生物被膜形成中起关键作用。PslD 为 Psl 多糖的转运蛋白,由256个氨基酸构成,生物信息学分析揭示其有 N 端信号肽且为一次跨膜蛋白。分离纯化完整跨膜蛋白需要去垢剂的作用,去垢剂种类繁多且性质不一,研究设计了一套筛选溶解 PslD 的去垢剂的方案。通过抗组氨酸标签的 Western blot 分析,n-Decyl-β-D-maltopyranoside(DM), n-Decyl-N,N-dimethylamine-N-oxide(DDAO)及 n-Dodecyl-N,N-dimethylamine-N-oxide(LDAO)被认为溶解 PslD的效率较高。通过改变总蛋白与去垢剂比例,进一步优化了去垢剂的溶解条件,即8 mg/ mL

  13. Extracellular electron transfer mechanisms between microorganisms and minerals

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Liang; Dong, Hailiang; Reguera, Gemma; Beyenal, Haluk; Lu, Anhuai; Liu, Juan; Yu, Han-Qing; Fredrickson, James K.

    2016-08-30

    Electrons can be transferred from microorganisms to multivalent metal ions that are associated with minerals and vice versa. As the microbial cell envelope is neither physically permeable to minerals nor electrically conductive, microorganisms have evolved strategies to exchange electrons with extracellular minerals. In this Review, we discuss the molecular mechanisms that underlie the ability of microorganisms to exchange electrons, such as c-type cytochromes and microbial nanowires, with extracellular minerals and with microorganisms of the same or different species. Microorganisms that have extracellular electron transfer capability can be used for biotechnological applications, including bioremediation, biomining and the production of biofuels and nanomaterials.

  14. Evolutionary adaptations of biofilms infecting cystic fibrosis lungs promote mechanical toughness by adjusting polysaccharide production.

    Science.gov (United States)

    Kovach, Kristin; Davis-Fields, Megan; Irie, Yasuhiko; Jain, Kanishk; Doorwar, Shashvat; Vuong, Katherine; Dhamani, Numa; Mohanty, Kishore; Touhami, Ahmed; Gordon, Vernita D

    2017-01-01

    Biofilms are communities of microbes embedded in a matrix of extracellular polymeric substances, largely polysaccharides. Multiple types of extracellular polymeric substances can be produced by a single bacterial strain. The distinct polymer components of biofilms are known to provide chemical protection, but little is known about how distinct extracellular polysaccharides may also protect biofilms against mechanical stresses such as shear or phagocytic engulfment. Decades-long infections of Pseudomonas. aeruginosa biofilms in the lungs of cystic fibrosis patients are natural models for studies of biofilm fitness under pressure from antibiotics and the immune system. In cystic fibrosis infections, production of the extracellular polysaccharide alginate has long been known to increase with time and to chemically protect biofilms. More recently, it is being recognized that chronic cystic fibrosis infections also evolve to increase production of another extracellular polysaccharide, Psl; much less is known about Psl's protective benefits to biofilms. We use oscillatory bulk rheology, on biofilms grown from longitudinal clinical isolates and from genetically-manipulated lab strains, to show that increased Psl stiffens biofilms and increases biofilm toughness, which is the energy cost to cause the biofilm to yield mechanically. Further, atomic force microscopy measurements reveal greater intercellular cohesion for higher Psl expression. Of the three types of extracellular polysaccharides produced by P. aeruginosa, only Psl increases the stiffness. Stiffening by Psl requires CdrA, a protein that binds to mannose groups on Psl and is a likely cross-linker for the Psl components of the biofilm matrix. We compare the elastic moduli of biofilms to the estimated stresses exerted by neutrophils during phagocytosis, and infer that increased Psl could confer a mechanical protection against phagocytic clearance.

  15. Signal Inhibitory Receptor on Leukocytes-1 Limits the Formation of Neutrophil Extracellular Traps, but Preserves Intracellular Bacterial Killing

    NARCIS (Netherlands)

    Van Avondt, Kristof; van der Linden, Maarten; Naccache, Paul H; Egan, David A; Meyaard, Linde

    2016-01-01

    In response to microbial invasion, neutrophils release neutrophil extracellular traps (NETs) to trap and kill extracellular microbes. Alternatively, NET formation can result in tissue damage in inflammatory conditions and may perpetuate autoimmune disease. Intervention strategies that are aimed at m

  16. Polysaccharides of the red algae.

    Science.gov (United States)

    Usov, Anatolii I

    2011-01-01

    Red algae (Rhodophyta) are known as the source of unique sulfated galactans, such as agar, agarose, and carrageenans. The wide practical uses of these polysaccharides are based on their ability to form strong gels in aqueous solutions. Gelling polysaccharides usually have molecules built up of repeating disaccharide units with a regular distribution of sulfate groups, but most of the red algal species contain more complex galactans devoid of gelling ability because of various deviations from the regular structure. Moreover, several red algae may contain sulfated mannans or neutral xylans instead of sulfated galactans as the main structural polysaccharides. This chapter is devoted to a description of the structural diversity of polysaccharides found in the red algae, with special emphasis on the methods of structural analysis of sulfated galactans. In addition to the structural information, some data on the possible use of red algal polysaccharides as biologically active polymers or as taxonomic markers are briefly discussed. Copyright © 2011 Elsevier Inc. All rights reserved.

  17. Why Were Polysaccharides Necessary?

    Science.gov (United States)

    Tolstoguzov, Vladimir

    2004-12-01

    The main idea of this paper is that the primordial soup may be modelled by food systems whose structure-property relationship is based on non-specific interactions between denatured biopolymers. According to the proposed hypothesis, polysaccharides were the first biopolymers that decreased concentration of salts in the primordial soup, `compatibilised' and drove the joint evolution of proto-biopolymers. Synthesis of macromolecules within the polysaccharide-rich medium could have resulted in phase separation of the primordial soup and concentration of the polypeptides and nucleic acids in the dispersed phase particles. The concentration of proto-biopolymer mixtures favoured their cross-linking in hybrid supermacromolecules of conjugates. The cross-linking of proto-biopolymers could occur by hydrophobic, electrostatic interactions, H-bonds due to freezing aqueous mixed biopolymer dispersions and/or by covalent bonds due to the Maillard reaction. Cross-linking could have increased the local concentration of chemically different proto-biopolymers, fixed their relative positions and made their interactions reproducible. Attractive-repulsive interactions between cross-linked proto-biopolymer chains could develop pairing of the monomer units, improved chemical stability (against hydrolysis) and led to their mutual catalytic activity and coding. Conjugates could probably evolve to the first self-reproduced entities and then to specialized cellular organelles. Phase separation of the primordial soup with concentration of conjugates in the dispersed particles has probably resulted in proto-cells.

  18. Enzymatic Modification of Plant Cell Wall Polysaccharides

    DEFF Research Database (Denmark)

    Øbro, Jens; Hayashi, Takahisa; Mikkelsen, Jørn Dalgaard

    2011-01-01

    for sustainable processes that replace chemical treatments with white biotechnology. Plants can contribute significantly to this sustainable process by producing plant or microbialenzymes in planta that are necessary for plant cell wall modification or total degradation. This will give rise to superior food......Plant cell walls are intricate structures with remarkable properties, widely used in almost every aspect of our life. Cell walls consist largely of complex polysaccharides and there is often a need for chemical and biochemical processing before industrial use. There is an increasing demand...... fibres, hydrocolloids, paper,textile, animal feeds or biofuels. Classical microbial-based fermentation systems could in the future face serious competition from plant-based expression systems for enzyme production. Plant expressed enzymes can either be targeted to specific cellular compartments...

  19. Enzymatic Modification of Plant Cell Wall Polysaccharides

    DEFF Research Database (Denmark)

    Øbro, Jens; Hayashi, Takahisa; Mikkelsen, Jørn Dalgaard

    2011-01-01

    fibres, hydrocolloids, paper,textile, animal feeds or biofuels. Classical microbial-based fermentation systems could in the future face serious competition from plant-based expression systems for enzyme production. Plant expressed enzymes can either be targeted to specific cellular compartments......Plant cell walls are intricate structures with remarkable properties, widely used in almost every aspect of our life. Cell walls consist largely of complex polysaccharides and there is often a need for chemical and biochemical processing before industrial use. There is an increasing demand...... for sustainable processes that replace chemical treatments with white biotechnology. Plants can contribute significantly to this sustainable process by producing plant or microbialenzymes in planta that are necessary for plant cell wall modification or total degradation. This will give rise to superior food...

  20. Polysaccharides: Molecular and Supramolecular Structures. Terminology.

    NARCIS (Netherlands)

    Heinze, Thomas; Petzold-Welcke, Katrin; Dam, van J.E.G.

    2012-01-01

    This chapter summarises important issues
    about the molecular and supramolecular structure
    of polysaccharides. It describes the terminology
    of polysaccharides systematically. The
    polysaccharides are divided regarding the
    molecular structures in glucans, polyoses,
    polysaccharid

  1. Membrane biofouling in a wastewater nitrification reactor: Microbial succession from autotrophic colonization to heterotrophic domination.

    Science.gov (United States)

    Lu, Huijie; Xue, Zheng; Saikaly, Pascal; Nunes, Suzana P; Bluver, Ted R; Liu, Wen-Tso

    2016-01-01

    Membrane biofouling is a complex process that involves bacterial adhesion, extracellular polymeric substances (EPS) excretion and utilization, and species interactions. To obtain a better understanding of the microbial ecology of biofouling process, this study conducted rigorous, time-course analyses on the structure, EPS and microbial composition of the fouling layer developed on ultrafiltration membranes in a nitrification bioreactor. During a 14-day fouling event, three phases were determined according to the flux decline and microbial succession patterns. In Phase I (0-2 days), small sludge flocs in the bulk liquid were selectively attached on membrane surfaces, leading to the formation of similar EPS and microbial community composition as the early biofilms. Dominant populations in small flocs, e.g., Nitrosomonas, Nitrobacter, and Acinetobacter spp., were also the major initial colonizers on membranes. In Phase II (2-4 d), fouling layer structure, EPS composition, and bacterial community went through significant changes. Initial colonizers were replaced by fast-growing and metabolically versatile heterotrophs (e.g., unclassified Sphingobacteria). The declining EPS polysaccharide to protein (PS:PN) ratios could be correlated well with the increase in microbial community diversity. In Phase III (5-14 d), heterotrophs comprised over 90% of the community, whereas biofilm structure and EPS composition remained relatively stable. In all phases, AOB and NOB were constantly found within the top 40% of the fouling layer, with the maximum concentrations around 15% from the top. The overall microbial succession pattern from autotrophic colonization to heterotrophic domination implied that MBR biofouling could be alleviated by forming larger bacterial flocs in bioreactor suspension (reducing autotrophic colonization), and by designing more specific cleaning procedures targeting dominant heterotrophs during typical filtration cycles. Copyright © 2015 Elsevier Ltd. All

  2. Membrane biofouling in a wastewater nitrification reactor: microbial succession from autotrophic colonization to heterotrophic domination

    KAUST Repository

    Lu, Huijie

    2015-10-22

    Membrane biofouling is a complex process that involves bacterial adhesion, extracellular polymeric substances (EPS) excretion and utilization, and species interactions. To obtain a better understanding of the microbial ecology of biofouling process, this study conducted rigorous, time-course analyses on the structure, EPS and microbial composition of the fouling layer developed on ultrafiltration membranes in a nitrification bioreactor. During a 14-day fouling event, three phases were determined according to the flux decline and microbial succession patterns. In Phase I (0-2 days), small sludge flocs in the bulk liquid were selectively attached on membrane surfaces, leading to the formation of similar EPS and microbial community composition as the early biofilms. Dominant populations in small flocs, e.g., Nitrosomonas, Nitrobacter, and Acinetobacter spp., were also the major initial colonizers on membranes. In Phase II (2-4 d), fouling layer structure, EPS composition, and bacterial community went through significant changes. Initial colonizers were replaced by fast-growing and metabolically versatile heterotrophs (e.g., unclassified Sphingobacteria). The declining EPS polysaccharide to protein (PS:PN) ratios could be correlated well with the increase in microbial community diversity. In Phase III (5-14 d), heterotrophs comprised over 90% of the community, whereas biofilm structure and EPS composition remained relatively stable. In all phases, AOB and NOB were constantly found within the top 40% of the fouling layer, with the maximum concentrations around 15% from the top. The overall microbial succession pattern from autotrophic colonization to heterotrophic domination implied that MBR biofouling could be alleviated by forming larger bacterial flocs in bioreactor suspension (reducing autotrophic colonization), and by designing more specific cleaning procedures targeting dominant heterotrophs during typical filtration cycles.

  3. A comparison of plate assay methods for detecting extracellular cellulase and xylanase activity.

    Science.gov (United States)

    Meddeb-Mouelhi, Fatma; Moisan, Jessica Kelly; Beauregard, Marc

    2014-11-01

    Identification of microorganisms for the production of carbohydrolytic enzymes is extremely important given the increased demand for these enzymes in many industries. To this end, dye-polysaccharide interactions which provide a visual indication of polymer hydrolysis (clear zones or halos) have been used for decades. For the detection of extracellular cellulase or xylanase activity many laboratories use Gram's iodine as the chromogenic dye, as it is a more rapid initial screening method compared to the use of other dyes. Here, we compared Gram's iodine and Congo red as indicators of polysaccharide hydrolysis. We attempted to detect cellulase activity using carboxymethylcellulose, and xylanase activity using birchwood xylan, in fourteen uncharacterized bacteria isolated from wood chips. Our results indicate that Gram's iodine may lead to identification of false positives in a typical screening protocol and that Congo red allows for avoidance of such pitfall. Congo red allowed detection of cellulase activity from live microbial colonies but not Gram's iodine. To confirm this, detection of enzymatic activity was also assessed using cell-free enzyme preparations. Congo red was found to be reliable in detecting cellulase activity with isolated enzymes preparations. Under the same conditions, neither of these dyes detected xylanase activity, despite independent evidence of xylanase activity for one of the preparations. We detected xylanase activity for this particular enzyme preparation using a coloured derivative of xylan (Remazol Brillant Blue R-xylan adduct) that respond to xylan hydrolysis. Our results suggest that methods that rely on interactions between a dye (Congo red or Gram's iodine) and a polymeric substrate (carboxymethylcellulose or birchwood xylan) for indirect detection of hydrolysis may require the use of relevant controls and independent confirmation of enzymatic activities.

  4. Screening of polysaccharides for preparation of α-amylase conjugate to enhance stability and storage life.

    Science.gov (United States)

    Jadhav, Swati B; Singhal, Rekha S

    2013-02-15

    Nine polysaccharides differing in structure and chemical nature were screened for their ability to conjugate with α-amylase by covalent binding for enhancing the thermal and pH stability of α-amylase. Among these polysaccharides, agar, dextran, pectin and xanthan showed better results but dextran stood out among all the polysaccharide for providing both thermal and pH stability to α-amylase. α-Amylase conjugated with agar, dextran, pectin and xanthan showed antimicrobial property with added preservative (0.2% sodium benzoate) in liquid formulation of α-amylase challenged with Bacillus subtilis and Escherichia coli. Dextran was the only polysaccharide which showed significant reduction of microbial growth of challenged organisms and aerobic flora without any preservative added. Aerobic flora could flourish well in the liquid α-amylase, but low temperature (4 °C), dextran, and preservative showed synergistic effect in efficiently increasing the storage life of liquid α-amylase.

  5. Metabolic engineering of microbes for oligosaccharide and polysaccharide synthesis

    Directory of Open Access Journals (Sweden)

    Chen Rachel

    2006-07-01

    Full Text Available Abstract Metabolic engineering has recently been embraced as an effective tool for developing whole-cell biocatalysts for oligosaccharide and polysaccharide synthesis. Microbial catalysts now provide a practical means to derive many valuable oligosaccharides, previously inaccessible through other methods, in sufficient quantities to support research and clinical applications. The synthesis process based upon these microbes is scalable as it avoids expensive starting materials. Most impressive is the high product concentrations (up to 188 g/L achieved through microbe-catalyzed synthesis. The overall cost for selected molecules has been brought to a reasonable range (estimated $ 30–50/g. Microbial synthesis of oligosaccharides and polysaccharides is a carbon-intensive and energy-intensive process, presenting some unique challenges in metabolic engineering. Unlike nicotinamide cofactors, the required sugar nucleotides are products of multiple interacting pathways, adding significant complexity to the metabolic engineering effort. Besides the challenge of providing the necessary mammalian-originated glycosyltransferases in active form, an adequate uptake of sugar acceptors can be an issue when another sugar is necessary as a carbon and energy source. These challenges are analyzed, and various strategies used to overcome these difficulties are reviewed in this article. Despite the impressive success of the microbial coupling strategy, there is a need to develop a single strain that can achieve at least the same efficiency. Host selection and the manner with which the synthesis interacts with the central metabolism are two important factors in the design of microbial catalysts. Additionally, unlike in vitro enzymatic synthesis, product degradation and byproduct formation are challenges of whole-cell systems that require additional engineering. A systematic approach that accounts for various and often conflicting requirements of the synthesis holds

  6. Computer simulation and experimental study of the polysaccharide-polysaccharide interaction in the bacteria Azospirillum brasilense Sp245

    Science.gov (United States)

    Arefeva, Oksana A.; Kuznetsov, Pavel E.; Tolmachev, Sergey A.; Kupadze, Machammad S.; Khlebtsov, Boris N.; Rogacheva, Svetlana M.

    2003-09-01

    We have studied the conformational properties and molecular dynamics of polysaccharides by using molecular modeling methods. Theoretical and experimental results of polysaccharide-polysaccharide interactions are described.

  7. Physicochemical characteristics and microbial community evolution of biofilms during the start-up period in a moving bed biofilm reactor.

    Science.gov (United States)

    Zhu, Yan; Zhang, Yan; Ren, Hong-Qiang; Geng, Jin-Ju; Xu, Ke; Huang, Hui; Ding, Li-Li

    2015-03-01

    This study aimed to investigate biofilm properties evolution coupled with different ages during the start-up period in a moving bed biofilm reactor system. Physicochemical characteristics including adhesion force, extracellular polymeric substances (EPS), morphology as well as volatile solid and microbial community were studied. Results showed that the formation and development of biofilms exhibited four stages, including (I) initial attachment and young biofilm formation, (II) biofilms accumulation, (III) biofilm sloughing and updating, and (IV) biofilm maturation. During the whole start-up period, adhesion force was positively and significantly correlated with the contents of EPS, especially the content of polysaccharide. In addition, increased adhesion force and EPS were beneficial for biofilm retention. Gram-negative bacteria mainly including Sphaerotilus, Zoogloea and Haliscomenobacter were predominant in the initial stage. Actinobacteria was beneficial to resist sloughing. Furthermore, filamentous bacteria were dominant in maturation biofilm. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Submerged cultivation of Ganoderma lucidum biomass and immunostimulatory effects of fungal polysaccharides.

    Science.gov (United States)

    Berovic, Marin; Habijanic, Jozica; Zore, Irena; Wraber, Branka; Hodzar, Damjan; Boh, Bojana; Pohleven, Franc

    2003-06-12

    Original Ganoderma lucidum strain MZKI G97 isolated from Slovenian forests was cultivated in a liquid substrate based on potato dextrose and olive oil. The influences of inoculum and oxygen partial pressure in batch and fed batch cultivation in a 10-l laboratory stirred tank reactor were studied. Fungal biomass was found to be oxygen and shear sensible. Using a 17% (wet weight) 6 days old vegetative inoculum, 9.6 g l(-1) of dry biomass in batch cultivation and 15.2 g l(-1) in fed batch process were obtained. Extracellular (9.6 g l(-1)) and intracellular (6.3 g l(-1)) polysaccharide fractions were isolated. Extracellular polysaccharide fraction and four intracellular polysaccharide fractions were obtained. Polysaccharides were further separated by ion-exchange, gel and affinity chromatography. The isolated polysaccharides were mainly beta-D-glucanes. Immunostimulatory effects of isolates were tested on induction of cytokine (tumour necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma)) synthesis in primary cultures of human peripheral blood mononuclear cells (PBMC) isolated from a buffy coat. The TNF-alpha inducing activity is comparable with romurtide, which has been used as a supporting therapy in cancer patients treated with radiotherapy and/or chemotherapy.

  9. Microbially induced separation of quartz from hematite using sulfate reducing bacteria.

    Science.gov (United States)

    Prakasan, M R Sabari; Natarajan, K A

    2010-07-01

    Cells and metabolic products of Desulfovibrio desulfuricans were successfully used to separate quartz from hematite through environmentally benign microbially induced flotation. Bacterial metabolic products such as extracellular proteins and polysaccharides were isolated from both unadapted and mineral-adapted bacterial metabolite and their basic characteristics were studied in order to get insight into the changes brought about on bioreagents during adaptation. Interaction between bacterial cells and metabolites with minerals like hematite and quartz brought about significant surface-chemical changes on both the minerals. Quartz was rendered more hydrophobic, while hematite became more hydrophilic after biotreatment. The predominance of bacterial polysaccharides on interacted hematite and of proteins on quartz was responsible for the above surface-chemical changes, as attested through adsorption studies. Surface-chemical changes were also observed on bacterial cells after adaptation to the above minerals. Selective separation of quartz from hematite was achieved through interaction with quartz-adapted bacterial cells and metabolite. Mineral-specific proteins secreted by quartz-adapted cells were responsible for conferment of hydrophobicity on quartz resulting in enhanced separation from hematite through flotation.

  10. Viscoelastic Properties of Extracellular Polymeric Substances Can Strongly Affect Their Washing Efficiency from Reverse Osmosis Membranes.

    Science.gov (United States)

    Ferrando Chavez, Diana Lila; Nejidat, Ali; Herzberg, Moshe

    2016-09-01

    The role of the viscoelastic properties of biofouling layers in their removal from the membrane was studied. Model fouling layers of extracellular polymeric substances (EPS) originated from microbial biofilms of Pseudomonas aeruginosa PAO1 differentially expressing the Psl polysaccharide were used for controlled washing experiments of fouled RO membranes. In parallel, adsorption experiments and viscoelastic modeling of the EPS layers were conducted in a quartz crystal microbalance with dissipation (QCM-D). During the washing stage, as shear rate was elevated, significant differences in permeate flux recovery between the three different EPS layers were observed. According to the amount of organic carbon remained on the membrane after washing, the magnitude of Psl production provides elevated resistance of the EPS layer to shear stress. The highest flux recovery during the washing stage was observed for the EPS with no Psl. Psl was shown to elevate the layer's shear modulus and shear viscosity but had no effect on the EPS adhesion to the polyamide surface. We conclude that EPS retain on the membrane as a result of the layer viscoelastic properties. These results highlight an important relation between washing efficiency of fouling layers from membranes and their viscoelastic properties, in addition to their adhesion properties.

  11. Light-driven oxidation of polysaccharides by photosynthetic pigments and a metalloenzyme

    DEFF Research Database (Denmark)

    Cannella, David; Möllers, K. Benedikt; Frigaard, Niels-Ulrik;

    2016-01-01

    Oxidative processes are essential for the degradation of plant biomass. A class of powerful and widely distributed oxidative enzymes, the lytic polysaccharide monooxygenases (LPMOs), oxidize the most recalcitrant polysaccharides and require extracellular electron donors. Here we investigated...... and hemicellulose. LPMO enzymes and pigment derivatives common in the environment of plant-degrading organisms thus form a highly reactive and stable light-driven system increasing the turnover rate and versatility of LPMOs. This light-driven system may find applications in biotechnology and chemical processing....

  12. Extracellular DNA is required for root tip resistance to fungal infection.

    Science.gov (United States)

    Wen, Fushi; White, Gerard J; VanEtten, Hans D; Xiong, Zhongguo; Hawes, Martha C

    2009-10-01

    Plant defense involves a complex array of biochemical interactions, many of which occur in the extracellular environment. The apical 1- to 2-mm root tip housing apical and root cap meristems is resistant to infection by most pathogens, so growth and gravity sensing often proceed normally even when other sites on the root are invaded. The mechanism of this resistance is unknown but appears to involve a mucilaginous matrix or "slime" composed of proteins, polysaccharides, and detached living cells called "border cells." Here, we report that extracellular DNA (exDNA) is a component of root cap slime and that exDNA degradation during inoculation by a fungal pathogen results in loss of root tip resistance to infection. Most root tips (>95%) escape infection even when immersed in inoculum from the root-rotting pathogen Nectria haematococca. By contrast, 100% of inoculated root tips treated with DNase I developed necrosis. Treatment with BAL31, an exonuclease that digests DNA more slowly than DNase I, also resulted in increased root tip infection, but the onset of infection was delayed. Control root tips or fungal spores treated with nuclease alone exhibited normal morphology and growth. Pea (Pisum sativum) root tips incubated with [(32)P]dCTP during a 1-h period when no cell death occurs yielded root cap slime containing (32)P-labeled exDNA. Our results suggest that exDNA is a previously unrecognized component of plant defense, an observation that is in accordance with the recent discovery that exDNA from white blood cells plays a key role in the vertebrate immune response against microbial pathogens.

  13. Extraction, Characterization, and Molecular Weight Determination of Senna tora (L. Seed Polysaccharide

    Directory of Open Access Journals (Sweden)

    Harshal A. Pawar

    2015-01-01

    Full Text Available The objective of the present work was extraction of polysaccharide from Senna tora L. seed and its characterization as a pharmaceutical excipient. Polysaccharide extraction was based on mechanical separation of the endosperm of seeds of Senna tora, water dissolution, centrifugation, and precipitation with acetone. Standard procedures were used to study the viscosity, micromeritic properties, and microbial bioburden. Accelerated stability study was carried out on isolated polysaccharide for six months at 40°C/75 RH as per ICH guidelines. The gum obtained from S. tora seeds was an amorphous free flowing odourless powder with dull brown colour (yield = 35% w/w. The bulk density, tapped density, and angle of repose data reveal that S. tora gum possesses good flow property. The intrinsic viscosity obtained was 1.568 dL/g. The average molecular weight of purified S. tora gum was found to be 198 kDa by intrinsic viscosity method. The results indicated that viscosity of gum solution increases with increase in temperature. FTIR study revealed the absence of degradation or decomposition of polysaccharide at accelerated stability conditions for six months. It has been concluded that extracted polysaccharide can be used as pharmaceutical excipient in terms of flow behavior, microbial properties, and stability.

  14. Sulfation patterns determine cellular internalization of heparin-like polysaccharides.

    Science.gov (United States)

    Raman, Karthik; Mencio, Caitlin; Desai, Umesh R; Kuberan, Balagurunathan

    2013-04-01

    Heparin is a highly sulfated polysaccharide that serves biologically relevant roles as an anticoagulant and anticancer agent. While it is well-known that modification of heparin's sulfation pattern can drastically influence its ability to bind growth factors and other extracellular molecules, very little is known about the cellular uptake of heparin and the role sulfation patterns serve in affecting its internalization. In this study, we chemically synthesized several fluorescently labeled heparins consisting of a variety of sulfation patterns. These polysaccharides were thoroughly characterized using anion exchange chromatography and size exclusion chromatography. Subsequently, we utilized flow cytometry and confocal imaging to show that sulfation patterns differentially affect the amount of heparin uptake in multiple cell types. This study provides the first comprehensive analysis of the effect of sulfation pattern on the cellular internalization of heparin or heparan sulfate like polysaccharides. The results of this study expand current knowledge regarding heparin internalization and provide insights into developing more effective heparin-based drug conjugates for applications in intracellular drug delivery.

  15. A Look inside the Listeria monocytogenes Biofilms Extracellular Matrix

    Directory of Open Access Journals (Sweden)

    Angelo Colagiorgi

    2016-07-01

    Full Text Available Listeria monocytogenes is a foodborne pathogen able to persist in food industry and is responsible for a severe illness called listeriosis. The ability of L. monocytogenes to persist in environments is due to its capacity to form biofilms that are a sessile community of microorganisms embedded in a self-produced matrix of extracellular polymeric substances (EPS’s. In this review, we summarized recent efforts performed in order to better characterize the polymeric substances that compose the extracellular matrix (ECM of L. monocytogenes biofilms. EPS extraction and analysis led to the identification of polysaccharides, proteins, extracellular DNA, and other molecules within the listerial ECM. All this knowledge will be useful for increasing food protection, suggesting effective strategies for the minimization of persistence of L. monocytogenes in food industry environments.

  16. Sulfur activation-related extracellular proteins of Acidithiobacillus ferrooxidans

    Institute of Scientific and Technical Information of China (English)

    ZHANG Cheng-gui; ZHANG Rui-yong; XIA Jin-lan; ZHANG Qian; NIE Zhen-yuan

    2008-01-01

    The fractions of the extracellular proteins of Acidithiobacillus ferrooxidans grown on two different energy substrates,elemental sulfur and ferrous sulfate,were selectively prepared with hot water treatment and distinctly shown by two-dimensional gel electrophoresis.Some protein spots with apparently higher abundance in sulfur energy substrate than in ferrous sulfate energy substrate were identified by using MALDI-TOF/TOF.Based on peptide mass fingerprints and bioinformatical analysis,the extracellular proteins were classified according to their functions as conjugal transfer protein,pilin,vacJ lipoprotein,polysaccharide deacetylase family protein,Ser/Thr protein phosphatase family protein and hypothetical proteins.Several extracellular proteins were found abundant in thiol groups and with CXXC functional motif,these proteins may be directly involved in the sulfur activation by use of their thiol group (Pr-SH) to bond the elemental sulfur.

  17. PREPARATION AND PROPERTIES OF EXTRACELLULAR BIOPOLYMER FLOCCULANT

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The biopolymer flocculant (named PS-2) producing by Pseudomonas fluorescens was investigated. The PS-2 had high efficiency with small dosage, when dealing with kaolin suspension, formed larger floc, with big sedimentation rate, over a wide range of temperatures. Distributing of flocculating activity test showed that the biopolymer flocculant was an extracellular product. The composition analysis of purified biopolymer flocculant showed that it composed mainly of polysaccharide and nucleic acid. The content of polysaccharide was 86.7%, which determined by using phenol-vitriol method, and the content of nucleic acid was 7.8%, which determined by UV absorption method. The biopolymer flocculant as a powder form showed much better stability than that as a supernatant. The character of biopolymer flocculant was stable even it was heated to 100℃ when it in acidic condition. The optimal conditions to flocculate kaolin suspension were as follows: pH 8~12, flocculant dosage 1mL/L, and Ca2+ as the optimal cation.

  18. PREPARATION AND PROPERTIES OF EXTRACELLULAR BIOPOLYMER FLOCCULANT

    Institute of Scientific and Technical Information of China (English)

    LI Chunxiang; LIU Binbin; XIONG Jinshui; YAN Jingchun

    2007-01-01

    The biopolymer flocculant (named PS-2) producing by Pseudomonas fluorescens was investigated. The PS-2 had high efficiency with small dosage, when dealing with kaolin suspension,formed larger floc, with big sedimentation rate, over a wide range of temperatures. Distributing of flocculating activity test showed that the biopolymer flocculant was an extracellular product. The composition analysis of purified biopolymer flocculant showed that it composed mainly of polysaccharide and nucleic acid. The content of polysaccharide was 86.7%, which determined by using phenol-vitriol method, and the content of nucleic acid was 7.8%, which determined by UV absorption method. The biopolymer flocculant as a powder form showed much better stability than that as a supernatant. The character of biopolymer flocculant was stable even it was heated to 100 ℃ when it in acidic condition. The optimal conditions to flocculate kaolin suspension were as follows:pH 8~12, flocculant dosage 1mL/L, and Ca2+ as the optimal cation.

  19. Film forming microbial biopolymers for commercial applications--a review.

    Science.gov (United States)

    Vijayendra, S V N; Shamala, T R

    2014-12-01

    Microorganisms synthesize intracellular, structural and extracellular polymers also referred to as biopolymers for their function and survival. These biopolymers play specific roles as energy reserve materials, protective agents, aid in cell functioning, the establishment of symbiosis, osmotic adaptation and support the microbial genera to function, adapt, multiply and survive efficiently under changing environmental conditions. Viscosifying, gelling and film forming properties of these have been exploited for specific significant applications in food and allied industries. Intensive research activities and recent achievements in relevant and important research fields of global interest regarding film forming microbial biopolymers is the subject of this review. Microbial polymers such as pullulan, kefiran, bacterial cellulose (BC), gellan and levan are placed under the category of exopolysaccharides (EPS) and have several other functional properties including film formation, which can be used for various applications in food and allied industries. In addition to EPS, innumerable bacterial genera are found to synthesis carbon energy reserves in their cells known as polyhydroxyalkanoates (PHAs), microbial polyesters, which can be extruded into films with excellent moisture and oxygen barrier properties. Blow moldable biopolymers like PHA along with polylactic acid (PLA) synthesized chemically in vitro using lactic acid (LA), which is produced by LA bacteria through fermentation, are projected as biodegradable polymers of the future for packaging applications. Designing and creating of new property based on requirements through controlled synthesis can lead to improvement in properties of existing polysaccharides and create novel biopolymers of great commercial interest and value for wider applications. Incorporation of antimicrobials such as bacteriocins or silver and copper nanoparticles can enhance the functionality of polymer films especially in food packaging

  20. Extracellular DNA contributes to dental biofilm formation: an ex vivo study

    DEFF Research Database (Denmark)

    Schlafer, Sebastian; Meyer, Rikke Louise; Dige, Irene;

    The extracellular matrix of dental biofilms plays an important role during caries development. It increases the mechanical stability of the biofilm, it prevents desiccation, it serves as a reservoir for nutrients and it contributes to the long-term preservation of acidic microenvironments. Research...... on the biofilm matrix in the field of dentistry has focused mainly on the synthesis, structure and function of extracellular polysaccharides. In recent years, studies conducted on biofilms from other habitats have shown that the presence of extracellular DNA contributes to biofilm formation and stability...

  1. Extracellular Polymers in Granular Sludge from Different Upflow Anaerobic Sludge Blanket (UASB) Reactors

    DEFF Research Database (Denmark)

    Schmidt, Jens Ejbye; Ahring, Birgitte Kiær

    1994-01-01

    Thermal extraction was used to quantify extracellular polymers (ECP) in granules from anaerobic upflow reactors. The optimal time for extraction was determined as the time needed before the intracellular material gives a significant contribution to the extracted extracellular material due to cell...... of an upflow anaerobic sludge blanket reactor from a sugar-containing waste-water to a synthetic waste-water containing acetate, propionate and butyrate resulted in a decrease in both the protein and polysaccharide content and an increase in the lipid content of the extracellular material. Furthermore...

  2. O-acetylation of Plant Cell Wall Polysaccharides

    Directory of Open Access Journals (Sweden)

    Sascha eGille

    2012-01-01

    Full Text Available Plant cell walls are composed of structurally diverse polymers, many of which are O-acetylated. How plants O-acetylate wall polymers and what its function is remained elusive until recently, when two protein families were identified in the model plant Arabidopsis that are involved in the O-acetylation of wall polysaccharides – the reduced wall acetylation (RWA and the trichome birefringence-like (TBL proteins. This review discusses the role of these two protein families in polysaccharide O-acetylation and outlines the differences and similarities of polymer acetylation mechanisms in plants, fungi, bacteria and mammals. Members of the TBL protein family had been shown to impact pathogen resistance, freezing tolerance, and cellulose biosynthesis. The connection of TBLs to polysaccharide O-acetylation thus gives crucial leads into the biological function of wall polymer O-acetylation.From a biotechnological point understanding the O-acetylation mechanism is important as acetyl-substituents inhibit the enzymatic degradation of wall polymers and released acetate can be a potent inhibitor in microbial fermentations, thus impacting the economic viability of e.g. lignocellulosic based biofuel production.

  3. Polysaccharide-modified synthetic polymeric biomaterials.

    Science.gov (United States)

    Baldwin, Aaron D; Kiick, Kristi L

    2010-01-01

    This review presents an overview of polysaccharide-conjugated synthetic polymers and their use in tissue-engineered scaffolds and drug-delivery applications. This topic will be divided into four categories: (1) polymeric materials modified with non-mammalian polysaccharides such as alginate, chitin, and dextran; (2) polymers modified with mammalian polysaccharides such as hyaluronan, chondroitin sulfate, and heparin; (3) multi-polysaccharide-derivatized polymer conjugate systems; and (4) polymers containing polysaccharide-mimetic molecules. Each section will discuss relevant conjugation techniques, analysis, and the impact of these materials as micelles, particles, or hydrogels used in in-vitro and in-vivo biomaterial applications. (c) 2010 Wiley Periodicals, Inc.

  4. Preliminary Study on the Liquid Fermentation and Polysaccharide Production of Ganoderma%灵芝液体发酵及产多糖的初步研究

    Institute of Scientific and Technical Information of China (English)

    王新民; 李宇伟; 连瑞丽; 边传周; 王慧杰

    2007-01-01

    Strain of Ganoderma japonicum with higher mycelial growth rate and higher levels of polysaccharide production was selected from seven tested strains of Canoderma.The effects of nitrogen source ,carbon sourloe and metal ions on the polysaccharide production of selected Ganoderma japonicum were studied.The results showed that the optimum nitrogen source and carbon source were 2%and 0.2%,respoctively.And the existence of 0.2%Fe2+ could improve the polysaccharide yield significantly.The result of fermentor enlargement test indicated that extracellular crude polysaccharide content per 100 mL fermentation liquor and mycelium content yielded highest amount of 181.7 mg and 151.0 mg,respectively,with relatively mild change of pH value.It was concluded that fermentation in fermentor was more suitable for the secretion of Ganoderma polysaccharide.

  5. Enhanced Production of Polysaccharide Through the Overexpression of Homologous Uridine Diphosphate Glucose Pyrophosphorylase Gene in a Submerged Culture of Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Higher Basidiomycetes).

    Science.gov (United States)

    Ji, Sen-Lin; Liu, Rui; Ren, Meng-Fei; Li, Huan-Jun; Xu, Jun-Wei

    2015-01-01

    This study aimed to improve polysaccharide production by engineering the biosynthetic pathway in Ganoderma lucidum through the overexpression of the homologous UDP glucose pyrophosphorylase (UGP) gene. The effects of UGP gene overexpression on intracellular polysaccharide (IPS) content, extracellular polysaccharide (EPS) production, and transcription levels of 3 genes encoding the enzymes involved in polysaccharide biosynthesis, including phosphoglucomutase (PGM), UGP, and α-1,3-glucan synthase (GLS), were investigated. The maximum IPS content and EPS production in G. lucidum overexpressing the UGP gene were 24.32 mg/100 mg dry weight and 1.66 g/L, respectively, which were higher by 42% and 36% than those of the wild-type strain. The transcription levels of PGM, UGP, and GLS were up-regulated by 1.6, 2.6, and 2.4-fold, respectively, in the engineered strain, suggesting that increased polysaccharide biosynthesis may result from a higher expression of those genes.

  6. Submerged cultivation of Ganoderma lucidum and the effects of its polysaccharides on the production of human cytokines TNF-α, IL-12, IFN-γ, IL-2, IL-4, IL-10 and IL-17.

    Science.gov (United States)

    Habijanic, Jožica; Berovic, Marin; Boh, Bojana; Plankl, Mojca; Wraber, Branka

    2015-01-25

    An original strain of Ganoderma lucidum (W.Curt.:Fr.) Lloyd, MZKI G97 isolated from Slovenian habitats was grown by a submerged liquid substrate cultivation in a laboratory stirred tank reactor. Five fractions of extracellular and cell-wall polysaccharides were obtained by extraction, ethanol precipitation, and purification by ion-exchange, gel and affinity chromatography. The capacity of isolated polysaccharide fractions to induce innate inflammatory cytokines, and to modulate cytokine responses of activated lymphocytes was investigated. Human peripheral blood mononuclear cells (PBMC) were activated in vitro with polysaccharide fractions, in order to induce innate inflammatory cytokines: tumor necrosis factor alpha (TNF-α), interleukin (IL) 12 and interferon gamma (IFN-γ). For the immunomodulation capacity, polysaccharide fractions were cultured with ionomycine and phorbol myristate acetate (IONO+PMA) activated PBMC, and the concentrations of induced IL-2, IL-4, IFN-γ, IL-10 and IL-17 were measured. The results showed that polysaccharides from G. lucidum induced moderate to high amounts of innate inflammatory cytokines. Fungal cell-wall polysaccharides were stronger innate inflammatory cytokines inducers, while extracellular polysaccharides demonstrated a higher capacity to modulate cytokine responses of IONO+PMA induced production of IL-17. The results indicate that G. lucidum polysaccharides enhance Th1 response with high levels of IFN-γ and IL-2, and display low to no impact on IL-4 production. A similar pattern was observed at regulatory cytokine IL-10. All of the polysaccharide fractions tested induced IL-17 production at different concentration levels.

  7. Correlative light/electron microscopy for the investigation of microbial mats from Black Sea Cold Seeps.

    Science.gov (United States)

    Wrede, Christoph; Heller, Christina; Reitner, Joachim; Hoppert, Michael

    2008-05-01

    In several fields of cell biology, correlative microscopy is applied to compare the structure of objects at high resolution under the electron microscope with low resolution light microscopy images of the same sample. It is, however, difficult to prepare samples and marker systems that are applicable for both microscopic techniques for the same specimen at the same time. In our studies, we used microbial mats from Cold Seep communities for a simple and rapid correlative microscopy method. The mats consist of bacterial and archaeal microorganisms, coupling reverse methanogenesis to the reduction of sulfate. The reverse methanogenic pathway also generates carbonates that precipitate inside the mat and may be the main reason for the formation of a microbial reef. The mat shows highly differentiated aggregates of various organisms, tightly interconnected by extracellular polysaccharides. In order to investigate the role of EPS as adhesive mucilage for the biofilm and as a precipitation matrix for carbonate minerals, samples were embedded in a hydrophilic resin (Lowicryl K4 M). Sections were suitable for light as well as electron microscopy in combination with lectins, either labeled with a fluorescent marker or with colloidal gold. This allows lectin mapping at low resolution for light microscopy in direct comparison with a highly resolved electron microscopic image.

  8. Extracellular calcium sensing and extracellular calcium signaling

    Science.gov (United States)

    Brown, E. M.; MacLeod, R. J.; O'Malley, B. W. (Principal Investigator)

    2001-01-01

    The cloning of a G protein-coupled extracellular Ca(2+) (Ca(o)(2+))-sensing receptor (CaR) has elucidated the molecular basis for many of the previously recognized effects of Ca(o)(2+) on tissues that maintain systemic Ca(o)(2+) homeostasis, especially parathyroid chief cells and several cells in the kidney. The availability of the cloned CaR enabled the development of DNA and antibody probes for identifying the CaR's mRNA and protein, respectively, within these and other tissues. It also permitted the identification of human diseases resulting from inactivating or activating mutations of the CaR gene and the subsequent generation of mice with targeted disruption of the CaR gene. The characteristic alterations in parathyroid and renal function in these patients and in the mice with "knockout" of the CaR gene have provided valuable information on the CaR's physiological roles in these tissues participating in mineral ion homeostasis. Nevertheless, relatively little is known about how the CaR regulates other tissues involved in systemic Ca(o)(2+) homeostasis, particularly bone and intestine. Moreover, there is evidence that additional Ca(o)(2+) sensors may exist in bone cells that mediate some or even all of the known effects of Ca(o)(2+) on these cells. Even more remains to be learned about the CaR's function in the rapidly growing list of cells that express it but are uninvolved in systemic Ca(o)(2+) metabolism. Available data suggest that the receptor serves numerous roles outside of systemic mineral ion homeostasis, ranging from the regulation of hormonal secretion and the activities of various ion channels to the longer term control of gene expression, programmed cell death (apoptosis), and cellular proliferation. In some cases, the CaR on these "nonhomeostatic" cells responds to local changes in Ca(o)(2+) taking place within compartments of the extracellular fluid (ECF) that communicate with the outside environment (e.g., the gastrointestinal tract). In others

  9. Dynamic remodeling of microbial biofilms by functionally distinct exopolysaccharides.

    Science.gov (United States)

    Chew, Su Chuen; Kundukad, Binu; Seviour, Thomas; van der Maarel, Johan R C; Yang, Liang; Rice, Scott A; Doyle, Patrick; Kjelleberg, Staffan

    2014-08-05

    Biofilms are densely populated communities of microbial cells protected and held together by a matrix of extracellular polymeric substances. The structure and rheological properties of the matrix at the microscale influence the retention and transport of molecules and cells in the biofilm, thereby dictating population and community behavior. Despite its importance, quantitative descriptions of the matrix microstructure and microrheology are limited. Here, particle-tracking microrheology in combination with genetic approaches was used to spatially and temporally study the rheological contributions of the major exopolysaccharides Pel and Psl in Pseudomonas aeruginosa biofilms. Psl increased the elasticity and effective cross-linking within the matrix, which strengthened its scaffold and appeared to facilitate the formation of microcolonies. Conversely, Pel reduced effective cross-linking within the matrix. Without Psl, the matrix becomes more viscous, which facilitates biofilm spreading. The wild-type biofilm decreased in effective cross-linking over time, which would be advantageous for the spreading and colonization of new surfaces. This suggests that there are regulatory mechanisms to control production of the exopolysaccharides that serve to remodel the matrix of developing biofilms. The exopolysaccharides were also found to have profound effects on the spatial organization and integration of P. aeruginosa in a mixed-species biofilm model of P. aeruginosa-Staphylococcus aureus. Pel was required for close association of the two species in mixed-species microcolonies. In contrast, Psl was important for P. aeruginosa to form single-species biofilms on top of S. aureus biofilms. Our results demonstrate that Pel and Psl have distinct physical properties and functional roles during biofilm formation. Importance: Most bacteria grow as biofilms in the environment or in association with eukaryotic hosts. Removal of biofilms that form on surfaces is a challenge in clinical

  10. Polysaccharide matrices used in 3D in vitro cell culture systems.

    Science.gov (United States)

    Diekjürgen, Dorina; Grainger, David W

    2017-10-01

    Polysaccharides comprise a diverse class of polymeric materials with a history of proven biocompatibility and continual use as biomaterials. Recent focus on new matrices appropriate for three-dimensional (3D) cell culture offers new opportunities to apply polysaccharides as extracellular matrix mimics. However, chemical and structural bases for specific cell-polysaccharide interactions essential for their utility as 3-D cell matrices are not well defined. This review describes how these naturally sourced biomaterials satisfy several key properties for current 3D cell culture needs and can also be synthetically modified or blended with additional components to tailor their cell engagement properties. Beyond their benign interactions with many cell types in cultures, their economical and high quality sourcing, optical clarity for ex situ analytical interrogation and in situ gelation represent important properties of these polymers for 3D cell culture applications. Continued diversification of their versatile glycan chemistry, new bio-synthetic sourcing strategies and elucidation of new cell-specific properties are attractive to expand the polysaccharide polymer utility for cell culture needs. Many 3D cell culture priorities are addressed with the portfolio of polysaccharide materials available and under development. This review provides a critical analysis of their properties, capabilities and challenges in 3D cell culture applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Polysaccharide production by submerged and solid-state cultures from several medicinal higher Basidiomycetes.

    Science.gov (United States)

    Montoya, Sandra; Sanchez, Oscar Julian; Levin, Laura

    2013-01-01

    Polysaccharides produced by microorganisms represent an industrially unexploited market. An important number of polysaccharides have been isolated from fungi, especially mushrooms, with many interesting biological functions, such as antitumor, hypoglycemic, and immunostimulating activities. In the search of new sources of fungal polysaccharides, the main goal of this research was to test the ability of several species of basidiomycetes, among them various edible mushrooms, to produce both extracellular polysaccharides (EPSs) and intracellular polysaccharides (IPSs). Among 10 species screened for production of EPSs in submerged cultures with glucose, soy oil, and yeast extract, the best results were obtained with Ganoderma lucidum (0.79 g/L EPS) and Pleurotus ostreatus (0.75 g/L EPS). Agitation strongly improved EPS production in most of the studied strains. Eight of 10 species assayed successfully developed basidiomes during synthetic "bag-log" cultivation on a substrate consisting of oak sawdust and corn bran. This work describes for the first time the environmental factors required for fruiting of 4 species under such conditions: Schizophyllum commune, Ganoderma applanatum, Trametes versicolor, and T. trogii. IPSs were extracted from the carpophores. The IPS content of the carpophores varied from 1.4% (G. applanatum) up to 5.5% and 6% in G. lucidum and Grifola frondosa, respectively.

  12. Isolation and chemical characterization of dissolved and particulate polysaccharides in Mikawa Bay

    Science.gov (United States)

    Sakugawa, Hiroshi; Handa, Nobuhiko

    1985-05-01

    Isolation and chemical elucidation of dissolved and particulate polysaccharides in seawater were conducted. The water samples were collected in Mikawa Bay, Japan during a red tide bloom of the dinoflagellate, Prorocentrum minimum. Dissolved polysaccharides were concentrated from 5-101 of seawater with dialysis followed by separation by gel flitration, and isolation by ethanol precipitation. A heteropolysaccharide consisting of glucose, galactose, mannose, xylose, arabinose, fucose and rhamnose and a glucan were isolated from the polysaccharide component having a molecular weight more than 4,000 Dalton and were characterized by several chemical analyses. The heteropolysaccharide is a mucilaginous polysaccharide having a highly branched structure and a molecular weight of 10 4-5 × 10 6 Daltons and probably contains a sulfate half ester: the glucan is a polysaccharide with β-1,3- and 1,6-linkages (chrysolaminaran type). Concentrations of these were respectively ca. 20 and 67 μg l -1 at 1 m, and 2 and 26 μg l -1 at 6 m. A similar heteropolysaccharide was found in the boiling water extract of the particulate matter, while β-glucan was isolated in a much less purified form than the seawater β-glucan. In addition, a large amount of β-1,4 glucan was found in the strong alkali extract of the particulate matter, indicating that this glucan must be a cell wall polysaccharide derived from phytoplankton. These results strongly suggest that the heteropolysaccharide and chrysolaminaran type polysaccharide dissolved in seawater were derived from water soluble carbohydrates of phytoplankton through extracellular release or cell lysis.

  13. Extracellular enzyme activity and biogeochemical cycling in restored prairies

    Science.gov (United States)

    Lynch, L.; Hernandez, D.; Schade, J. D.

    2011-12-01

    Winter microbial activity in mid-latitude prairie ecosystems is thermally sensitive and significantly influenced by snow depth. Snow insulates the soil column facilitating microbial processing of complex organic substrates. Previous studies in forests and tundra ecosystems suggest patterns of substrate utilization and limitation are seasonal; above freezing, soil microbes access fresh litter inputs and sugar exudates from plant roots, while under frozen condition they recycle nutrients incorporated in microbial biomass. In order to liberate nutrients required for carbon degradation, soil microbes invest energy in the production of extracellular enzymes that cleave monomers from polymer bonds. The inverse relationship between relative enzyme abundance and substrate availability makes enzyme assays a useful proxy to assess changes in resources over time. Our objective in this study was to assess patterns in microbial biomass, nutrient availability, and extracellular enzyme activity in four snow exclosure sites over a seven-month period. Over the past three years, we have maintained a snow removal experiment on two restored prairies in central Minnesota. In each prairie, snow was continuously removed annually from two 4 x 4 m plots by shoveling after each snow event. Extractable C, N and P, and microbial C, N and P in soil samples were measured in samples collected from these snow removal plots, as well as in adjacent unmanipulated prairie control plots. Pools of C, N, and P were estimated using standard extraction protocols, and microbial pools were estimated using chloroform fumigation direct extraction (CFDE). We conducted fluorometric extracellular enzyme assays (EEA) to assess how the degradation potential of cellulose (cellobiohydrolase, CBH), protein (leucine aminopeptidase, LAP), and phosphate esters (phosphatase, PHOS) changed seasonally. Microbial C and N declined between October and June, while microbial P declined during the fall and winter, but increased

  14. Sucrose release from polysaccharide gels.

    Science.gov (United States)

    Nishinari, Katsuyoshi; Fang, Yapeng

    2016-05-18

    Sucrose release from polysaccharide gels has been studied extensively because it is expected to be useful in understanding flavour release from solid foods and to find a new processing method which produces more palatable and healthier foods. We provide an overview of the release of sucrose and other sugars from gels of agar and related polysaccharides. The addition of sucrose to agar solutions leads to the increase in transparency of the resulting gels and the decrease in syneresis, which is attributed to the decrease in mesh size in gels. The syneresis occurring in the quiescent condition and fluid release induced by compression is discussed. The relationship between the sugar release and the structural, rheological and thermal properties of gels is also discussed. Finally, the future research direction is proposed.

  15. Polysaccharide-Based Micelles for Drug Delivery

    Directory of Open Access Journals (Sweden)

    Nan Zhang

    2013-05-01

    Full Text Available Delivery of hydrophobic molecules and proteins has been an issue due to poor bioavailability following administration. Thus, micelle carrier systems are being investigated to improve drug solubility and stability. Due to problems with toxicity and immunogenicity, natural polysaccharides are being explored as substitutes for synthetic polymers in the development of new micelle systems. By grafting hydrophobic moieties to the polysaccharide backbone, self-assembled micelles can be readily formed in aqueous solution. Many polysaccharides also possess inherent bioactivity that can facilitate mucoadhesion, enhanced targeting of specific tissues, and a reduction in the inflammatory response. Furthermore, the hydrophilic nature of some polysaccharides can be exploited to enhance circulatory stability. This review will highlight the advantages of polysaccharide use in the development of drug delivery systems and will provide an overview of the polysaccharide-based micelles that have been developed to date.

  16. Fingerprinting of hydroxyl radical-attacked polysaccharides by N-isopropyl-2-aminoacridone labelling.

    Science.gov (United States)

    Vreeburg, Robert A M; Airianah, Othman B; Fry, Stephen C

    2014-10-15

    Hydroxyl radicals (•OH) cause non-enzymic scission of polysaccharides in diverse biological systems. Such reactions can be detrimental (e.g. causing rheumatic and arthritic diseases in mammals) or beneficial (e.g. promoting the softening of ripening fruit, and biomass saccharification). Here we present a method for documenting •OH action, based on fluorescent labelling of the oxo groups that are introduced as glycosulose residues when •OH attacks polysaccharides. The method was tested on several polysaccharides, especially pectin, after treatment with Fenton reagents. 2-Aminoacridone plus cyanoborohydride reductively aminated the oxo groups in treated polysaccharides; the product was then reacted with acetone plus cyanoborohydride, forming a stable tertiary amine with the carbohydrate linked to N-isopropyl-2-aminoacridone (pAMAC). Digestion of labelled pectin with 'Driselase' yielded several fluorescent products which on electrophoresis and HPLC provided a useful 'fingerprint' indicating •OH attack. The most diagnostic product was a disaccharide conjugate of the type pAMAC·UA-GalA (UA=unspecified uronic acid), whose UA-GalA bond was Driselase-resistant (product 2A). 2A was clearly distinguishable from GalA-GalA-pAMAC (disaccharide labelled at its reducing end), which was digestible to GalA-pAMAC. The methodology is applicable, with appropriate enzymes in place of Driselase, for detecting natural and artificial •OH attack in diverse plant, animal and microbial polysaccharides.

  17. Polysaccharides As Safer Release Systems For Agrochemicals

    OpenAIRE

    Campos E.V.R.; de Oliveira J.L.; Fraceto L.F.; Singh B

    2014-01-01

    International audience; Agrochemicals are used to improve the production of crops. Conventional formulations of agrochemicals can contaminate the environment, in particular in the case of intensive cropping. Hence, there is a need for controlled-release formulations of agrochemicals such as polysaccharides to reduce pollution and health hazards. Natural polysaccharides are hydrophilic, biodegradable polymers. This article reviews the use of polysaccharides in the form of micro- and nanopartic...

  18. Retention Behaviors of Uronic Acid-containing Polysaccharides and Neutral Polysaccharides in HPGPC

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The chromatographic behaviors of several uronic acid-containing polysaccharides and neutral polysaccharides were investigated in HPGPC for the first time. The effects of sample concentration and ionic strength of mobile phase on retention time were studied. The mechanism for the effects on Mw determination results of polysaccharides by HPGPC was also discussed.

  19. Advances on Bioactive Polysaccharides from Medicinal Plants.

    Science.gov (United States)

    Xie, Jian-Hua; Jin, Ming-Liang; Morris, Gordon A; Zha, Xue-Qiang; Chen, Han-Qing; Yi, Yang; Li, Jing-En; Wang, Zhi-Jun; Gao, Jie; Nie, Shao-Ping; Shang, Peng; Xie, Ming-Yong

    2016-07-29

    In recent decades, the polysaccharides from the medicinal plants have attracted a lot of attention due to their significant bioactivities, such as anti-tumor activity, antioxidant activity, anticoagulant activity, antidiabetic activity, radioprotection effect, anti-viral activity, hypolipidemic and immunomodulatory activities, which make them suitable for medicinal applications. Previous studies have also shown that medicinal plant polysaccharides are non-toxic and show no side effects. Based on these encouraging observations, most researches have been focusing on the isolation and identification of polysaccharides, as well as their bioactivities. A large number of bioactive polysaccharides with different structural features and biological effects from medicinal plants have been purified and characterized. This review provides a comprehensive summary of the most recent developments in physiochemical, structural features and biological activities of bioactive polysaccharides from a number of important medicinal plants, such as polysaccharides from Astragalus membranaceus, Dendrobium plants, Bupleurum, Cactus fruits, Acanthopanax senticosus, Angelica sinensis (Oliv.) Diels, Aloe barbadensis Miller, and Dimocarpus longan Lour. Moreover, the paper has also been focused on the applications of bioactive polysaccharides for medicinal applications. Recent studies have provided evidence that polysaccharides from medicinal plants can play a vital role in bioactivities. The contents and data will serve as a useful reference material for further investigation, production, and application of these polysaccharides in functional foods and therapeutic agents.

  20. Extracellular Gd-CA

    DEFF Research Database (Denmark)

    Thomsen, Henrik S; Marckmann, Peter

    2008-01-01

    Until recently it was believed that extracellular gadolinium-based contrast agents were safe for both the kidneys and all other organs within the dose range up to 0.3 mmol/kg body weight. However, in 2006, it was demonstrated that some gadolinium-based contrast agents may trig the development of ...

  1. Microbial Flocculant for Nature Soda

    Energy Technology Data Exchange (ETDEWEB)

    Qin, Peiyong; Zhang, Tong; Chen, Cuixian

    2004-03-31

    Microbial flocculant for nature soda has been studied. Lactobacillus TRJ21, which was able to produce an excellent biopolymer flocculant for nature soda, was obtained in our lab. The microbial flocculant was mainly produced when the bacteria laid in stationary growth phase. Fructose or glucose, as carbon sources, were more favorable for the bacterial growth and flocculant production. The bacteria was able to use ammonium sulfate or Urea as nitrogen to produce flocculant, but was not able to use peptone effectively. High C/N ratio was more favorable to Lactobacillus TRJ21 growth and flocculant production than low C/N ratio. The biopolymer flocculant was mainly composed of polysaccharide and protein with a molecular weight 1.38x106 by gel permeation chromatography. It was able to be easily purified from the culture medium by acetone. Protein in the flocculant was tested for the flocculating activity ingredient by heating the flocculant.

  2. Sucrose fed-batch strategy enhanced biomass, polysaccharide, and ganoderic acids production in fermentation of Ganoderma lucidum 5.26.

    Science.gov (United States)

    Wei, Zhen-hua; Liu, Lianliang; Guo, Xiao-feng; Li, Yan-jun; Hou, Bao-chao; Fan, Qiu-ling; Wang, Kai-xiang; Luo, Yingdi; Zhong, Jian-jiang

    2016-01-01

    Ganoderma, as a Chinese traditional medicine, has multiple bioactivities. However, industrial production was limited due to low yield during Ganoderma fermentation. In this work, sucrose was found to greatly enhance intracellular polysaccharide (IPS) content and specific extracellular polysaccharide (EPS) production rate. The mechanism was studied by analyzing the activities of enzymes related to polysaccharide biosynthesis. The results revealed that sucrose regulated the activities of phosphoglucomutase and phosphoglucose isomerase. When glucose and sucrose mixture was used as carbon source, biomass, polysaccharide and ganoderic acids (GAs) production was greatly enhanced. A sucrose fed-batch strategy was developed in 10-L bioreactor, and was scaled up to 300-L bioreactor. The biomass, EPS and IPS production was 25.5, 2.9 and 4.8 g/L, respectively, which was the highest biomass and IPS production in pilot scale. This study provides information for further understanding the regulation mechanism of Ganoderma polysaccharide biosynthesis. It demonstrates that sucrose fed-batch is a useful strategy for enhancing Ganoderma biomass, polysaccharide and GAs production.

  3. Different Types of Diatom-Derived Extracellular Polymeric Substances Drive Changes in Heterotrophic Bacterial Communities from Intertidal Sediments

    Science.gov (United States)

    Bohórquez, Julio; McGenity, Terry J.; Papaspyrou, Sokratis; García-Robledo, Emilio; Corzo, Alfonso; Underwood, Graham J. C.

    2017-01-01

    Intertidal areas support extensive diatom-rich biofilms. Such microphytobenthic (MPB) diatoms exude large quantities of extracellular polymeric substances (EPS) comprising polysaccharides, glycoproteins and other biopolymers, which represent a substantial carbon pool. However, degradation rates of different EPS components, and how they shape heterotrophic communities in sediments, are not well understood. An aerobic mudflat-sediment slurry experiment was performed in the dark with two different EPS carbon sources from a diatom-dominated biofilm: colloidal EPS (cEPS) and the more complex hot-bicarbonate-extracted EPS. Degradation rate constants determined over 9 days for three sediment fractions [dissolved organic carbon (DOC), total carbohydrates (TCHO), and (cEPS)] were generally higher in the colloidal-EPS slurries (0.105–0.123 d−1) compared with the hot-bicarbonate-extracted-EPS slurries (0.060–0.096 d−1). Addition of hot-bicarbonate-EPS resulted in large increases in dissolved nitrogen and phosphorous by the end of the experiment, indicating that the more complex EPS is an important source of regenerated inorganic nutrients. Microbial biomass increased ~4–6-fold over 9 days, and pyrosequencing of bacterial 16S rRNA genes revealed that the addition of both types of EPS greatly altered the bacterial community composition (from 0 to 9 days) compared to a control with no added EPS. Bacteroidetes (especially Tenacibaculum) and Verrucomicrobia increased significantly in relative abundance in both the hot-bicarbonate-EPS and colloidal-EPS treatments. These differential effects of EPS fractions on carbon-loss rates, nutrient regeneration and microbial community assembly improve our understanding of coastal-sediment carbon cycling and demonstrate the importance of diverse microbiota in processing this abundant pool of organic carbon. PMID:28289404

  4. Enhanced stability and dissolution of CuO nanoparticles by extracellular polymeric substances in aqueous environment

    Energy Technology Data Exchange (ETDEWEB)

    Miao, Lingzhan; Wang, Chao; Hou, Jun, E-mail: hhuhjyhj@126.com; Wang, Peifang; Ao, Yanhui; Li, Yi; Lv, Bowen; Yang, Yangyang; You, Guoxiang; Xu, Yi [Hohai University, Key Laboratory of Integrated Regulation and Resources Development on Shallow Lakes, Ministry of Education (China)

    2015-10-15

    Stability of engineered nanoparticles in aquatic environment is an essential parameter to evaluate their fate, bioavailability, and potential toxic effects toward living organisms. As CuO NPs enter the wastewater systems, they will encounter extracellular polymeric substances (EPS) from microbial community before directly interacting with bacterial cells. EPS may play an important role in affecting the stability and the toxicity of CuO NPs in aquatic environment. In this study, the influences of flocculent sludge-derived EPS, as well as model protein (BSA) and natural polysaccharides (alginate) on the dissolution kinetics and colloidal stability of CuO NPs were investigated. Results showed that the presence of NOMs strongly suppressed CuO NPs aggregation, confirmed by DLS, zeta potentials, and TEM analysis. The enhanced stability of CuO NPs in the presence of EPS and alginate were attributed to the electrostatic combined with steric repulsion, while the steric-hindrance effect may be the predominant mechanism retarding nano-CuO aggregation for BSA. Higher degrees of copper release were achieved with the increasing concentrations of NOMs. EPS are more effective than alginate and BSA in releasing copper, probably due to the abundant functional groups and the excellent metal-binding capacity. The ratio of free-Cu{sup 2+}/total dissolved Cu significantly decreased in the presence of EPS, indicating that EPS may affect the speciation and Cu bioavailability in aqueous environment. These results may be important for assessing the fate and transport behaviors of CuO NPs in the environment as well as for setting up usage regulation and treatment strategy.

  5. Starch-degrading polysaccharide monooxygenases.

    Science.gov (United States)

    Vu, Van V; Marletta, Michael A

    2016-07-01

    Polysaccharide degradation by hydrolytic enzymes glycoside hydrolases (GHs) is well known. More recently, polysaccharide monooxygenases (PMOs, also known as lytic PMOs or LPMOs) were found to oxidatively degrade various polysaccharides via a copper-dependent hydroxylation. PMOs were previously thought to be either GHs or carbohydrate binding modules (CBMs), and have been re-classified in carbohydrate active enzymes (CAZY) database as auxiliary activity (AA) families. These enzymes include cellulose-active fungal PMOs (AA9, formerly GH61), chitin- and cellulose-active bacterial PMOs (AA10, formerly CBM33), and chitin-active fungal PMOs (AA11). These PMOs significantly boost the activity of GHs under industrially relevant conditions, and thus have great potential in the biomass-based biofuel industry. PMOs that act on starch are the latest PMOs discovered (AA13), which has expanded our perspectives in PMOs studies and starch degradation. Starch-active PMOs have many common structural features and biochemical properties of the PMO superfamily, yet differ from other PMO families in several important aspects. These differences likely correlate, at least in part, to the differences in primary and higher order structures of starch and cellulose, and chitin. In this review we will discuss the discovery, structural features, biochemical and biophysical properties, and possible biological functions of starch-active PMOs, as well as their potential application in the biofuel, food, and other starch-based industries. Important questions regarding various aspects of starch-active PMOs and possible economical driving force for their future studies will also be highlighted.

  6. Thermal studies on natural polysaccharide

    Institute of Scientific and Technical Information of China (English)

    Sunil B Bothara; Sudarshan Singh

    2012-01-01

    Objective: To characterize thermal property of natural gums obtained from the seeds of Diospyros melonoxylon(D. melonoxylon) Roxb, Buchanania lanzan (B. lanzan) spreng and Manilkara zapota (M. zapota) (Linn.) P. Royen syn. Methods: Natural gums were thermally characterized using differential scanning calorimetry (DSC), differential thermal analysis (DTA) and thermo-gravimetric analysis (TGA) under nitrogen atmosphere. Major thermal transitions as well as activation energies of the major decomposition stages were determined. Elemental analysis was performed in order to determine the composition of carbon, hydrogen, nitrogen and sulfur. Results: DSC traces indicated a major intense exothermic transition (around 200℃) followed by weaker exotherm(s). Thermogravimetric analysis showed two phase of weight loss. The first phase has minor weight loss in samples is attributed to the loss of adsorbed and structural water of biopolymers or due to desorption of moisture as hydrogen bound water to the saccharide structure. The second weight loss event may be attributed to the polysaccharide decomposition. The initial decomposition temperature (IDT) was calculated from thermograms obtained of TGA, seed Polysaccharide of D. melonoxylon (IDT 221.21℃), B. lanzan (IPDT 170.4℃) and M. zapota (IPDT 178.6℃) were obtained. According to the integral procedural decomposition temperature (IPDT) values calculated based on the TGA thermograms; D. melonoxylon (IPDT 563.3℃), B. lanzan (IPDT 598.1℃) and M. zapota (IPDT 600.6℃) were obtained respectively. The elemental analysis study shows that the isolated natural Polysaccharides consist of certain percentage of carbon, nitrogen, sulphur and hydrogen in all the gums. Conclusions: The results of the present investigation reveal that the natural gums are thermally stable and these gums can be used as release modifiers in various dosage forms.

  7. Pharmacological Action of Adenophora Polysaccharides

    Institute of Scientific and Technical Information of China (English)

    李泱; 李春红; 唐富天; 李新芳

    2004-01-01

    Adenophora polysaccharides (AP), is an active principle extracted from the root of Adenophorae Potaninii Korsh originated in Gansu Province and isolated with boiling water. AP is isolated and purified from the crude drug by DEAE-cellulose and Sephadex G-200 column, with a white powder and mean molecular weight of 8.3×104 , and [α]D20 of AP is + 68. AP is only composed of glucose judging from the analysis of it with patina chromatography (PC) and gas chromatography-mass spectrometer (GC-MS) methods.

  8. Bioactive polysaccharides and gut microbiome (abstract)

    Science.gov (United States)

    Many polysaccharides have shown the ability to reduce plasma cholesterol or postprandial glycemia. Viscosity in the small intestine seems to be required to slow glucose uptake. Cereal mixed linkage beta-glucans, psyllium, glucomannans, and other polysaccharides also seem to require higher molecula...

  9. Extracellular DNA: the tip of root defenses?

    Science.gov (United States)

    Hawes, Martha C; Curlango-Rivera, Gilberto; Wen, Fushi; White, Gerard J; Vanetten, Hans D; Xiong, Zhongguo

    2011-06-01

    This review discusses how extracellular DNA (exDNA) might function in plant defense, and at what level(s) of innate immunity this process might operate. A new role for extracellular factors in mammalian defense has been described in a series of studies. These studies reveal that cells including neutrophils, eosinophils, and mast cells produce 'extracellular traps' (ETs) consisting of histone-linked exDNA. When pathogens are attracted to such ETs, they are trapped and killed. When the exDNA component of ETs is degraded, trapping is impaired and resistance against invasion is reduced. Conversely, mutation of microbial genes encoding exDNases that degrade exDNA results in loss of virulence. This discovery that exDNases are virulence factors opens new avenues for disease control. In plants, exDNA is required for defense of the root tip. Innate immunity-related proteins are among a group of >100 proteins secreted from the root cap and root border cell populations. Direct tests revealed that exDNA also is rapidly synthesized and exported from the root tip. When this exDNA is degraded by the endonuclease DNase 1, root tip resistance to fungal infection is lost; when the polymeric structure is degraded more slowly, by the exonuclease BAL31, loss of resistance to fungal infection is delayed accordingly. The results suggest that root border cells may function in a manner analogous to that which occurs in mammalian cells.

  10. Production of a novel extracellular polysaccharide by Lactobacillus sake 0-1 and characterization of the polysaccharide

    NARCIS (Netherlands)

    Vliegenthart, J.F.G.; Berg, D.J.C. van den; Robijn, G.W.; Janssen, A.C.; Giuseppin, M.L.F.; Vreeker, R.; Kamerling, J.P.; Ledeboer, A.M.

    1995-01-01

    A novel exopolysaccharide (EPS) produced by Lactobacillus sake 0-1 (CBS 532.92) has been isolated and characterized. When the strain was grown on glucose, the produced EPS contained glucose and rhamnose in a molar ratio of 3:2 and the average molecular mass distribution (M(infm)) was determined at 6

  11. Polysaccharides in colon-specific drug delivery.

    Science.gov (United States)

    Sinha, V R; Kumria, R

    2001-08-14

    Natural polysaccharides are now extensively used for the development of solid dosage forms for delivery of drug to the colon. The rationale for the development of a polysaccharide based delivery system for colon is the presence of large amounts of polysaccharidases in the human colon as the colon is inhabited by a large number and variety of bacteria which secrete many enzymes e.g. beta-D-glucosidase, beta-D-galactosidase, amylase, pectinase, xylanase, beta-D-xylosidase, dextranase, etc. Various major approaches utilizing polysaccharides for colon-specific delivery are fermentable coating of the drug core, embedding of the drug in biodegradable matrix, formulation of drug-saccharide conjugate (prodrugs). A large number of polysaccharides have already been studied for their potential as colon-specific drug carrier systems, such as chitosan, pectin, chondroitin sulphate, cyclodextrin, dextrans, guar gum, inulin, amylose and locust bean gum. Recent efforts and approaches exploiting these polysaccharides in colon-specific drug delivery are discussed.

  12. Extracellular proteins limit the dispersal of biogenic nanoparticles

    Science.gov (United States)

    Moreau, J.W.; Weber, P.K.; Martin, M.C.; Gilbert, B.; Hutcheon, I.D.; Banfield, J.F.

    2007-01-01

    High-spatial-resolution secondary ion microprobe spectrometry, synchrotron radiation-based Fourier-transform infrared spectroscopy, and polyacrylamide gel analysis demonstrated the intimate association of proteins with spheroidal aggregates of biogenic zinc sulfide nanocrystals, an example of extracellular biomineralization. Experiments involving synthetic zinc sulfide nanoparticles and representative amino acids indicated a driving role for cysteine in rapid nanoparticle aggregation. These findings suggest that microbially derived extracellular proteins can limit the dispersal of nanoparticulate metal-bearing phases, such as the mineral products of bioremediation, that may otherwise be transported away from their source by subsurface fluid flow.

  13. Distinct roles of extracellular polymeric substances in Pseudomonas aeruginosa biofilm development

    DEFF Research Database (Denmark)

    Yang, Liang; Hu, Yifan; Liu, Yang

    2011-01-01

    Bacteria form surface attached biofilm communities as one of the most important survival strategies in nature. Biofilms consist of water, bacterial cells and a wide range of self‐generated extracellular polymeric substances (EPS). Biofilm formation is a dynamic self‐assembly process and several d...... polysaccharide is more important than Pel polysaccharide in P. aeruginosa PAO1 biofilm formation and antibiotic resistance. Our study thus suggests that different EPS materials play distinct roles during bacterial biofilm formation.......Bacteria form surface attached biofilm communities as one of the most important survival strategies in nature. Biofilms consist of water, bacterial cells and a wide range of self‐generated extracellular polymeric substances (EPS). Biofilm formation is a dynamic self‐assembly process and several...

  14. Immunomodulatory and Anti-IBDV Activities of the Polysaccharide AEX from Coccomyxa gloeobotrydiformis

    Science.gov (United States)

    Guo, Qiang; Shao, Qiang; Xu, Wenping; Rui, Lei; Sumi, Ryo; Eguchi, Fumio; Li, Zandong

    2017-01-01

    A number of polysaccharides have been reported to show immunomodulatory and antiviral activities against various animal viruses. AEX is a polysaccharide extracted from the green algae, Coccomyxa gloeobotrydiformis. The aim of this study was to examine the function of AEX in regulating the immune response in chickens and its capacity to inhibit the infectious bursal disease virus (IBDV), to gain an understanding of its immunomodulatory and antiviral ability. Here, preliminary immunological tests in vitro showed that the polysaccharide AEX can activate the chicken peripheral blood molecular cells’ (PBMCs) response by inducing the production of cytokines and NO, promote extracellular antigen presentation but negatively regulate intracellular antigen presentation in chicken splenic lymphocytes, and promote the proliferation of splenic lymphocytes and DT40 cells. An antiviral analysis showed that AEX repressed IBDV replication by the deactivation of viral particles or by interfering with adsorption in vitro and reduced the IBDV viral titer in the chicken bursa of Fabricius. Finally, in this study, when AEX was used as an adjuvant for the IBDV vaccine, specific anti-IBDV antibody (IgY, IgM, and IgA) titers were significantly decreased. These results indicate that the polysaccharide AEX may be a potential alternative approach for anti-IBDV therapy and an immunomodulator for the poultry industry. However, more experimentation is needed to find suitable conditions for it to be used as an adjuvant for the IBDV vaccine. PMID:28208594

  15. Natural polysaccharides promote chondrocyte adhesion and proliferation on magnetic nanoparticle/PVA composite hydrogels.

    Science.gov (United States)

    Hou, Ruixia; Nie, Lei; Du, Gaolai; Xiong, Xiaopeng; Fu, Jun

    2015-08-01

    This paper aims to investigate the synergistic effects of natural polysaccharides and inorganic nanoparticles on cell adhesion and growth on intrinsically cell non-adhesive polyvinyl alcohol (PVA) hydrogels. Previously, we have demonstrated that Fe2O3 and hydroxyapatite (nHAP) nanoparticles are effective in increasing osteoblast growth on PVA hydrogels. Herein, we blended hyaluronic acid (HA) and chondroitin sulfate (CS), two important components of cartilage extracellular matrix (ECM), with Fe2O3/nHAP/PVA hydrogels. The presence of these natural polyelectrolytes dramatically increased the pore size and the equilibrium swelling ratio (ESR) while maintaining excellent compressive strength of hydrogels. Chondrocytes were seeded and cultured on composite PVA hydrogels containing Fe2O3, nHAP and Fe2O3/nHAP hybrids and Fe2O3/nHAP with HA or CS. Confocal laser scanning microscopy (CLSM) and cell counting kit-8 (CCK-8) assay consistently confirmed that the addition of HA or CS promotes chondrocyte adhesion and growth on PVA and composite hydrogels. Particularly, the combination of HA and CS exhibited further promotion to cell adhesion and proliferation compared with any single polysaccharide. The results demonstrated that the magnetic composite nanoparticles and polysaccharides provided synergistic promotion to cell adhesion and growth. Such polysaccharide-augmented composite hydrogels may have potentials in biomedical applications.

  16. [The extraction, purification and assaying of Gynostemma pentaphyllum polysaccharides].

    Science.gov (United States)

    Song, Shu-liang; Tang, Jin-bao; Ji, Ai-guo; Liang, Hao; Zhu, Peng; Wang, Wei-li

    2006-06-01

    By orthogonal design, and considering extracting efficiency and cost, optimizing the extract method of Gynostemma pentaphyllum polysaccharides. We purified the crude Gynostemma pentaphyllum polysaccharides initially, and assayed the polysaccharides content of Gynostemma pentaphyllum polysccharides. The content of Gynostemma pentaphyllum polysaccharides was sigificantly higher than the predecessor. It would provide conditions for the deep exploitation of Gynostemma pentaphyllum.

  17. An exocellular polysaccharide and its interactions with proteins

    NARCIS (Netherlands)

    Tuinier, R.

    1999-01-01

    In the food industry polysaccharides are used as thickening or gelling agents. Polysaccharides are usually extracted from plants. Micro-organisms are also capable of excreting polysaccharides: exocellular polysaccharides (EPSs). In some cases EPSs are produced in-situ in food products, notably in ac

  18. Microbial biosurfactants and biodegradation.

    Science.gov (United States)

    Ward, Owen P

    2010-01-01

    Microbial biosurfactants are amphipathic molecules having typical molecular weights of 500-1500 Da, made up of peptides, saccharides or lipids or their combinations. In biodegradation processes they mediate solubilisation, mobilization and/or accession of hydrophobic substrates to microbes. They may be located on the cell surface or be secreted into the extracellular medium and they facilitate uptake of hydrophobic molecules through direct cellular contact with hydrophobic solids or droplets or through micellarisation. They are also involved in cell physiological processes such as biofilm formation and detachment, and in diverse biofilm associated processes such as wastewater treatment and microbial pathogenesis. The protection of contaminants in biosurfactants micelles may also inhibit uptake of contaminants by microbes. In bioremediation processes biosurfactants may facilitate release of contaminants from soil, but soils also tend to bind surfactants strongly which makes their role in contaminant desorption more complex. A greater understanding of the underlying roles played by biosurfactants in microbial physiology and in biodegradative processes is developing through advances in cell and molecular biology.

  19. Selective reactivity of monochloramine with extracellular matrix components affects the disinfection of biofilm and detached clusters.

    Science.gov (United States)

    Xue, Zheng; Lee, Woo Hyoung; Coburn, Kimberly M; Seo, Youngwoo

    2014-04-01

    The efficiency of monochloramine disinfection was dependent on the quantity and composition of extracellular polymeric substances (EPS) in biofilms, as monochloramine has a selective reactivity with proteins over polysaccharides. Biofilms with protein-based (Pseudomonas putida) and polysaccharide based EPS (Pseudomonas aeruginosa), as well as biofilms with varied amount of polysaccharide EPS (wild-type and mutant P. aeruginosa), were compared. The different reactivity of EPS components with monochloramine influenced disinfectant penetration, biofilm inactivation, as well as the viability of detached clusters. Monochloramine transport profiling measured by a chloramine-sensitive microelectrode revealed a broader diffusion boundary layer between bulk and biofilm surface in the P. putida biofilm compared to those of P. aeruginosa biofilms. The reaction with proteins in P. putida EPS multiplied both the time and the monochloramine mass required to achieve a full biofilm penetration. Cell viability in biofilms was also spatially influenced by monochloramine diffusion and reaction within biofilms, showing a lower survival in the surface section and a higher persistence in the middle section of the P. putida biofilm compared to the P. aeruginosa biofilms. While polysaccharide EPS promoted biofilm cell viability by obstructing monochloramine reactive sites on bacterial cells, protein EPS hindered monochloramine penetration by reacting with monochloramine and reduced its concentration within biofilms. Furthermore, the persistence of bacterial cells detached from biofilm (over 70% for P. putida and ∼40% for polysaccharide producing P. aeruginosa) suggested that currently recommended monochloramine residual levels may underestimate the risk of water quality deterioration caused by biofilm detachment.

  20. Sea Ice Microorganisms: Environmental Constraints and Extracellular Responses

    Directory of Open Access Journals (Sweden)

    Jody W. Deming

    2013-03-01

    Full Text Available Inherent to sea ice, like other high latitude environments, is the strong seasonality driven by changes in insolation throughout the year. Sea-ice organisms are exposed to shifting, sometimes limiting, conditions of temperature and salinity. An array of adaptations to survive these and other challenges has been acquired by those organisms that inhabit the ice. One key adaptive response is the production of extracellular polymeric substances (EPS, which play multiple roles in the entrapment, retention and survival of microorganisms in sea ice. In this concept paper we consider two main areas of sea-ice microbiology: the physico-chemical properties that define sea ice as a microbial habitat, imparting particular advantages and limits; and extracellular responses elicited in microbial inhabitants as they exploit or survive these conditions. Emphasis is placed on protective strategies used in the face of fluctuating and extreme environmental conditions in sea ice. Gaps in knowledge and testable hypotheses are identified for future research.

  1. Influence of O polysaccharides on biofilm development and outer membrane vesicle biogenesis in Pseudomonas aeruginosa PAO1.

    Science.gov (United States)

    Murphy, Kathleen; Park, Amber J; Hao, Youai; Brewer, Dyanne; Lam, Joseph S; Khursigara, Cezar M

    2014-04-01

    Pseudomonas aeruginosa is a common opportunistic human pathogen known for its ability to adapt to changes in its environment during the course of infection. These adaptations include changes in the expression of cell surface lipopolysaccharide (LPS), biofilm development, and the production of a protective extracellular exopolysaccharide matrix. Outer membrane vesicles (OMVs) have been identified as an important component of the extracellular matrix of P. aeruginosa biofilms and are thought to contribute to the development and fitness of these bacterial communities. The goal of this study was to examine the relationships between changes in the cell surface expression of LPS O polysaccharides, biofilm development, and OMV biogenesis in P. aeruginosa. We compared wild-type P. aeruginosa PAO1 with three chromosomal knockouts. These knockouts have deletions in the rmd, wbpM, and wbpL genes that produce changes in the expression of common polysaccharide antigen (CPA), O-specific antigen (OSA), or both. Our results demonstrate that changes in O polysaccharide expression do not significantly influence OMV production but do affect the size and protein content of OMVs derived from both CPA(-) and OSA(-) cells; these mutant cells also exhibited different physical properties from wild-type cells. We further examined biofilm growth of the mutants and determined that CPA(-) cells could not develop into robust biofilms and exhibit changes in cell morphology and biofilm matrix production. Together these results demonstrate the importance of O polysaccharide expression on P. aeruginosa OMV composition and highlight the significance of CPA expression in biofilm development.

  2. Economical evolution: microbes reduce the synthetic cost of extracellular proteins.

    Science.gov (United States)

    Smith, Daniel R; Chapman, Matthew R

    2010-08-24

    Protein evolution is not simply a race toward improved function. Because organisms compete for limited resources, fitness is also affected by the relative economy of an organism's proteome. Indeed, many abundant proteins contain relatively high percentages of amino acids that are metabolically less taxing for the cell to make, thus reducing cellular cost. However, not all abundant proteins are economical, and many economical proteins are not particularly abundant. Here we examined protein composition and found that the relative synthetic cost of amino acids constrains the composition of microbial extracellular proteins. In Escherichia coli, extracellular proteins contain, on average, fewer energetically expensive amino acids independent of their abundance, length, function, or structure. Economic pressures have strategically shaped the amino acid composition of multicomponent surface appendages, such as flagella, curli, and type I pili, and extracellular enzymes, including type III effector proteins and secreted serine proteases. Furthermore, in silico analysis of Pseudomonas syringae, Mycobacterium tuberculosis, Saccharomyces cerevisiae, and over 25 other microbes spanning a wide range of GC content revealed a broad bias toward more economical amino acids in extracellular proteins. The synthesis of any protein, especially those rich in expensive aromatic amino acids, represents a significant investment. Because extracellular proteins are lost to the environment and not recycled like other cellular proteins, they present a greater burden on the cell, as their amino acids cannot be reutilized during translation. We hypothesize that evolution has optimized extracellular proteins to reduce their synthetic burden on the cell.

  3. GEL PERMEATION CHROMATOGRAPHIC ANALYSIS OF LACQUER POLYSACCHARIDE

    Institute of Scientific and Technical Information of China (English)

    QIU Xingping; ZHANG Lina; DU Yumin; QIAN Baogong; LU Zaimin

    1992-01-01

    Ten fractionated samples of Chinese lacquer polysaccharide in aqueous 0.1M NaCl solution were studied by aqueous-phase gel permeation chromatography (GPC). The universal calibration, broad MWD calibration and corrected column dispersion were adopted to the analysis of GPC chromatograms of the polysaccharide. The molecular weights Mw, Mn and polydispersity index Mw/Mn obtained from GPC are in good agreement with the results of light scattering and membrane osmometry. It is verified that the universal calibration concept is applicable to the lacquer polysaccharide having a number of side chains.

  4. Polysaccharides: The “Click” Chemistry Impact

    Directory of Open Access Journals (Sweden)

    Romain Lucas

    2011-09-01

    Full Text Available Polysaccharides are complex but essential compounds utilized in many areas such as biomaterials, drug delivery, cosmetics, food chemistry or renewable energy. Modifications and functionalizations of such polymers are often necessary to achieve molecular structures of interest. In this area, the emergence of the “click” chemistry concept, and particularly the copper-catalyzed version of the Huisgen 1,3-dipolar cycloaddition reaction between terminal acetylenes and azides, had an impact on the polysaccharides chemistry. The present review summarizes the contribution of “click” chemistry in the world of polysaccharides.

  5. Nanocoating of titanium implant surfaces with organic molecules. Polysaccharides including glycosaminoglycans

    DEFF Research Database (Denmark)

    Gurzawska, Katarzyna Aleksandra; Svava, Rikke; Jørgensen, Niklas Rye;

    2012-01-01

    Long-term stability of titanium implants are dependent on a variety of factors. Nanocoating with organic molecules is one of the method used to improve osseointegration. Nanoscale modification of titanium implants affects surface properties, such as hydrophilicity, biochemical bonding capacity...... and roughness. This influences cell behaviour on the surface such as adhesion, proliferation and differentiation of cells as well as the mineralization of the extracellular matrix at the implant surfaces. The aim of the present systematic review was to describe organic molecules used for surface nanocoating...... nanocoatings. The included in vivo studies, showed improvement of bone interface reactions measured as increased Bone-to-Implant Contact length and Bone Mineral Density adjacent to the polysaccharide coated surfaces. Based on existing literature, surface modification with polysaccharide and glycosaminoglycans...

  6. Cryptococcus neoformans capsular polysaccharides form branched and complex filamentous networks viewed by high-resolution microscopy.

    Science.gov (United States)

    Araújo, Glauber R de S; Fontes, Giselle N; Leão, Daniela; Rocha, Gustavo Miranda; Pontes, Bruno; Sant'Anna, Celso; de Souza, Wanderley; Frases, Susana

    2016-01-01

    Cryptococcus neoformans is a fungal pathogen that causes life-threatening infections in immunocompromised individuals. Its main virulence factor is an extracellular polysaccharide capsule whose structure, assembly and dynamics remain poorly understood. In this study, we apply improved protocols for sample preparation and recently-developed scanning microscopy techniques to visualize the ultrastructure of the C. neoformans capsule at high-resolution (up to 1 nm) and improved structural preservation. Although most capsule structures in nature consist of linear polymers, we show here that the C. neoformans capsule is a 'microgel-like' structure composed of branched polysaccharides. Moreover, we imaged the capsule-to-cell wall link, which is formed by thin fibers that branch out of thicker capsule filaments, and have one end firmly embedded in the cell wall structure. Together, our findings provide compelling ultrastructural evidence for a branched and complex capsule conformation, which may have important implications for the biological activity of the capsule as a virulence factor.

  7. Getting to know the extracellular vesicle glycome.

    Science.gov (United States)

    Gerlach, Jared Q; Griffin, Matthew D

    2016-04-01

    Extracellular vesicles (EVs) are a diverse population of complex biological particles with diameters ranging from approximately 20 to 1000 nm. Tremendous interest in EVs has been generated following a number of recent, high-profile reports describing their potential utility in diagnostic, prognostic, drug delivery, and therapeutic roles. Subpopulations, such as exosomes, are now known to directly participate in cell-cell communication and direct material transfer. Glycomics, the 'omic' portion of the glycobiology field, has only begun to catalog the surface oligosaccharide and polysaccharide structures and also the carbohydrate-binding proteins found on and inside EVs. The EV glycome undoubtedly contains vital clues essential to better understanding the function, biogenesis, release and transfer of vesicles, however getting at this information is technically challenging and made even more so because of the small physical size of the vesicles and the typically minute yield from physiological-scale biological samples. Vesicle micro-heterogeneity which may be related to specific vesicle origins and functions presents a further challenge. A number of primary studies carried out over the past decade have turned up specific and valuable clues regarding the composition and roles of glycan structures and also glycan binding proteins involved EV biogenesis and transfer. This review explores some of the major EV glycobiological research carried out to date and discusses the potential implications of these findings across the life sciences.

  8. Extracellular Matrix Proteins

    Directory of Open Access Journals (Sweden)

    Linda Christian Carrijo-Carvalho

    2012-01-01

    Full Text Available Lipocalin family members have been implicated in development, regeneration, and pathological processes, but their roles are unclear. Interestingly, these proteins are found abundant in the venom of the Lonomia obliqua caterpillar. Lipocalins are β-barrel proteins, which have three conserved motifs in their amino acid sequence. One of these motifs was shown to be a sequence signature involved in cell modulation. The aim of this study is to investigate the effects of a synthetic peptide comprising the lipocalin sequence motif in fibroblasts. This peptide suppressed caspase 3 activity and upregulated Bcl-2 and Ki-67, but did not interfere with GPCR calcium mobilization. Fibroblast responses also involved increased expression of proinflammatory mediators. Increase of extracellular matrix proteins, such as collagen, fibronectin, and tenascin, was observed. Increase in collagen content was also observed in vivo. Results indicate that modulation effects displayed by lipocalins through this sequence motif involve cell survival, extracellular matrix remodeling, and cytokine signaling. Such effects can be related to the lipocalin roles in disease, development, and tissue repair.

  9. Pharmacological Action of Adenophora Polysaccharides

    Institute of Scientific and Technical Information of China (English)

    李泱; 李春红; 唐富天; 李新芳

    2004-01-01

    @@ Adenophora polysaccharides (AP), is an active principle extracted from the root of Adenophorae Potaninii Korsh originated in Gansu Province and isolated with boiling water(1). AP is isolated and purified from the crude drug by DEAE-cellulose and Sephadex G-200 column, with a white powder and mean molecular weight of 8.3 × 104 , and [α]D20of AP is + 68(1). AP is only composed of glucose judging from the analysis of it with patina chromatography (PC) and gas chromatography-mass spectrometer (GC-MS) methods.The methylation analysis showed that AP is composed of (1→6) linked glucose residues. The measure of nuclear magnetic resonance imaging (NMR) 1H NMR and 14C NMR techniques further proved that AP is α(l→6) linked by Dglucose. The structure of AP is as follows: -[→6]α-D-Glu(1-)n→ (2).

  10. Transport of Streptococcus pneumoniae capsular polysaccharide in MHC Class II tubules.

    Directory of Open Access Journals (Sweden)

    Tom Li Stephen

    2007-03-01

    Full Text Available Bacterial capsular polysaccharides are virulence factors and are considered T cell-independent antigens. However, the capsular polysaccharide Sp1 from Streptococcus pneumoniae serotype 1 has been shown to activate CD4(+ T cells in a major histocompatibility complex (MHC class II-dependent manner. The mechanism of carbohydrate presentation to CD4(+ T cells is unknown. We show in live murine dendritic cells (DCs that Sp1 translocates from lysosomal compartments to the plasma membrane in MHCII-positive tubules. Sp1 cell surface presentation results in reduction of self-peptide presentation without alteration of the MHCII self peptide repertoire. In DM-deficient mice, retrograde transport of Sp1/MHCII complexes resulting in T cell-dependent immune responses to the polysaccharide in vitro and in vivo is significantly reduced. The results demonstrate the capacity of a bacterial capsular polysaccharide antigen to use DC tubules as a vehicle for its transport as an MHCII/saccharide complex to the cell surface for the induction of T cell activation. Furthermore, retrograde transport requires the functional role of DM in self peptide-carbohydrate exchange. These observations open new opportunities for the design of vaccines against microbial encapsulated pathogens.

  11. Demonstration of pectic polysaccharides in cork cell wall from Quercus suber L.

    Science.gov (United States)

    Rocha, S M; Coimbra, M A; Delgadillo, I

    2000-06-01

    Scanning electron microscopy (SEM) and chemical analysis were used to observe the cell wall changes that occur in cork with "mancha amarela", when compared to a standard cork. To mimic the microbial attack exhibited in cork with mancha amarela, the standard cork was treated enzymatically with commercial pectinase and hemicellulase preparations. The tissues treated with pectinase were comparable with those attacked with mancha amarela. Both were composed by deformed and wrinkly cells and exhibited cell wall separation at the middle lamella level, which suggests solubilization/removal of the pectic polysaccharides. The cork cell wall material, prepared as alcohol-insoluble residue, was fractionated by hot water (Pect(H)()2(O)) and hot dilute acid (Pect(acid)). The relatively large amount of hexuronic acid and the occurrence of Ara in the SPect(H)()2(O) and SPect(acid) allow to confirm, as far as we know, for the first time the presence of pectic polysaccharides in the cell walls of cork from Quercus suber L. They accounted for ca. 1.5% of the cork and may consist of polymers with long side chains of arabinosyl residues. These polymers have to be taken into account in any realistic model of the cork cell wall. Cork with mancha amarela contained a smaller amount of pectic polysaccharides (ca. 0.5%), which confirms that the cellular separation observed by SEM is related to the degradation/removal of the middle lamella pectic polysaccharides.

  12. Serogroup quantitation of multivalent polysaccharide and polysaccharide-conjugate meningococcal vaccines from China.

    Science.gov (United States)

    Cook, Matthew C; Gibeault, Sabrina; Filippenko, Vasilisa; Ye, Qiang; Wang, Junzhi; Kunkel, Jeremy P

    2013-07-01

    The active components of most meningococcal vaccines are four antigenic serogroup capsular polysaccharides (A, C, Y, W135). The vaccines, monovalent or multivalent mixtures of either free polysaccharides or polysaccharides conjugated to antigenic carrier proteins, may be in liquid or lyophilised formulations, with or without excipients. Acid hydrolysis and chromatographic methods for serogroup quantitation, which were previously optimised and qualified using polysaccharide-based standards and a narrow range of real vaccines, are here challenged with multiple lots of a broad assortment of additional multivalent polysaccharide-based meningococcal vaccine products. Centrifugal filtration successfully removed all interfering lactose excipient without loss of polysaccharides to allow for the determination of Y and W135 serogroups. Replicate operations by three different analysts indicated high method reproducibility. Results indicated some lot-to-lot and product-to-product variations. However, all vaccines were within general specifications for each serogroup polysaccharide, with the exception of all lots of one polysaccharide vaccine - which by these methods were found to be deficient in the serogroup A component only. These robust techniques are very useful for the evaluation of antigen content and consistency of manufacture. The deformulation, hydrolysis and chromatographic methods may be adaptable for the evaluation of other types of polysaccharide-based vaccines.

  13. Functional advantages conferred by extracellular prokaryotic membrane vesicles.

    Science.gov (United States)

    Manning, Andrew J; Kuehn, Meta J

    2013-01-01

    The absence of subcellular organelles is a characteristic typically used to distinguish prokaryotic from eukaryotic cells. But recent discoveries do not support this dogma. Over the past 50 years, researchers have begun to appreciate and characterize Gram-negative bacterial outer membrane-derived vesicles and Gram-positive and archaeal membrane vesicles. These extracellular, membrane-bound organelles can perform a variety of functions, including binding and delivery of DNA, transport of virulence factors, protection of the cell from outer membrane targeting antimicrobials and ridding the cell of toxic envelope proteins. Here, we review the contributions of these extracellular organelles to prokaryotic physiology and compare these with the contributions of the bacterial interior membrane-bound organelles responsible for harvesting light energy and for generating magnetic crystals of heavy metals. Understanding the roles of these multifunctional extracellular vesicle organelles as microbial tools will help us to better realize the diverse interactions that occur in our polymicrobial world.

  14. Acathopanas Senticosus Polysaccharides: Effects on Growth Performance,Serum Immune Indices and Microbial Flora in Feces of Weaner Piglets%刺五加多糖对断奶仔猪生长性能、血清免疫指标及粪便微生物菌群的影响

    Institute of Scientific and Technical Information of China (English)

    杨侃侃; 边连全; 刘显军; 韩杰; 张飞

    2013-01-01

    本试验在基础饲粮中添加刺五加多糖,旨在研究其对断奶仔猪生长性能、血清免疫指标及粪便微生物菌群的影响.试验选用96头平均体重为(7.125 ±0.576) kg的28日龄断奶的“长白×约克夏”二元杂交仔猪,随机分为6个组,每组4个重复,每个重复4头猪.其中Ⅰ组为对照组,饲喂基础饲粮;Ⅱ~Ⅵ组饲喂在基础饲粮中分别添加150、300、500、800、1 000 mg/kg刺五加多糖的试验饲粮,试验期为21 d.结果表明:1)饲粮中添加刺五加多糖能够显著地提高试验全期(1 ~21 d)Ⅳ、V、Ⅵ组断奶仔猪的平均日增重(P<0.05),显著地降低试验15~21 d时Ⅲ、Ⅳ、V和Ⅵ组的料重比以及试验全期各试验组的腹泻率(P<0.05),但对试验各阶段的平均日采食量无显著影响(P>0.05).2)刺五加多糖能够显著提高试验全期Ⅲ、Ⅳ、V组断奶仔猪血清中免疫球蛋白A含量(P<0.05),显著提高试验l ~14d时Ⅲ、Ⅳ、V组和试验全期Ⅳ、V组免疫球蛋白G含量(P<0.05),显著提高试验全期V组免疫球蛋白M含量(P<0.05).3)刺五加多糖可以显著提高断奶仔猪粪便中乳酸杆菌和双歧杆菌的数量(P<0.05),显著降低大肠杆菌的数量(P<0.05).综合本试验各项指标,断奶仔猪饲粮中刺五加多糖最适宜添加量为800mg/kg.%This experiment was conducted to study the effects of Acathopanas senticosus polysaccharides (ASPS) on growth performance, serum immune indices and microbial flora in feces of weaner piglets. Ninety-six 28-day-old weaner piglets (Landrace × Yorkshire) with an average body weight of (7. 125 ±0. 576) kg were randomly allocated into 6 groups with 4 replicates in each group and 4 piglets in each replicate. Piglets in group Ⅰ (control) was fed a basal diet, and those in groups Ⅱ to Ⅵ were fed the basal diet supplemented with 150, 300, 500, 800 and 1 000 mg/kg ASPS, respectively. Piglets were raised for 21 days. The results showed as

  15. Role of Soil Microstructure in Microbially-mediated Drying Resistance

    Science.gov (United States)

    Cruz, B. C.; Shor, L. M.; Gage, D. J.

    2015-12-01

    The retention of soil moisture between rainfall or irrigation events is imperative to the productivity of terrestrial ecosystems. Amplified weather conditions are expected to result in widespread reduction in soil moisture. Extracellular polysaccharides (EPS) produced by soil bacteria have the ability to influence soil moisture by (i) retaining water directly within the hydrogel matrix, and (ii) promoting an aggregated soil structure. We have developed microfluidic devices that emulate realistic soil microstructures and enable direct observation of EPS production and drying resistance. The objective of this study was to compare moisture retention in emulated soil micromodels containing different soil microstructures. "Aggregated" devices contain a greater number of small (100 μm) pores, while "non-aggregated" devices contained more intermediate-sized (30-100 μm) pores. Particle-size distributions, similar to a sandy loam, were identical in both cases. Dilute suspensions of either of two strains of Sinorhizobium meliloti were introduced into replicate micromodels: one strain produced EPS ("EPS+") and the other did not produce EPS ("EPS-"). Loaded micromodels were equilibrated at saturated conditions, then dried at 83% RH for several days. Direct observation showed micro-scale patterns of air infiltration. The rate and extent of moisture loss was determined as a function of bacterial strain and microstructure aggregation state. Results showed devices loaded with EPS+ bacteria retained moisture longer than devices loaded with EPS- bacteria. Moisture retention by EPS+ bacteria was enhanced in aggregated versus non-aggregated microstructures. This work illustrates how moisture retention in soil is the result of microbial processes acting within pore-scale soil microstructures. Validated microfluidics-based approaches may help quantitatively link pore-scale phenomena to ecosystem function.

  16. Improved polysaccharide production in a submerged culture of Ganoderma lucidum by the heterologous expression of Vitreoscilla hemoglobin gene.

    Science.gov (United States)

    Li, Huan-Jun; Zhang, De-Huai; Yue, Tong-Hui; Jiang, Lu-Xi; Yu, Xuya; Zhao, Peng; Li, Tao; Xu, Jun-Wei

    2016-01-10

    Expression of Vitreoscilla hemoglobin (VHb) gene was used to improve polysaccharide production in Ganoderma lucidum. The VHb gene, vgb, under the control of the constitutive glyceraldehyde-3-phosphate dehydrogenase gene promoter was introduced into G. lucidum. The activity of expressed VHb was confirmed by the observation of VHb specific CO-difference spectrum with a maximal absorption at 419 nm for the transformant. The effects of VHb expression on intracellular polysaccharide (IPS) content, extracellular polysaccharide (EPS) production and transcription levels of three genes encoding the enzymes involved in polysaccharide biosynthesis, including phosphoglucomutase (PGM), uridine diphosphate glucose pyrophosphorylase (UGP), and β-1,3-glucan synthase (GLS), were investigated. The maximum IPS content and EPS production in the vgb-bearing G. lucidum were 26.4 mg/100mg dry weight and 0.83 g/L, respectively, which were higher by 30.5% and 88.2% than those of the wild-type strain. The transcription levels of PGM, UGP and GLS were up-regulated by 1.51-, 1.55- and 3.83-fold, respectively, in the vgb-bearing G. lucidum. This work highlights the potential of VHb to enhance G. lucidum polysaccharide production by large scale fermentation.

  17. The pel polysaccharide can serve a structural and protective role in the biofilm matrix of Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Kelly M Colvin

    2011-01-01

    Full Text Available Bacterial extracellular polysaccharides are a key constituent of the extracellular matrix material of biofilms. Pseudomonas aeruginosa is a model organism for biofilm studies and produces three extracellular polysaccharides that have been implicated in biofilm development, alginate, Psl and Pel. Significant work has been conducted on the roles of alginate and Psl in biofilm development, however we know little regarding Pel. In this study, we demonstrate that Pel can serve two functions in biofilms. Using a novel assay involving optical tweezers, we demonstrate that Pel is crucial for maintaining cell-to-cell interactions in a PA14 biofilm, serving as a primary structural scaffold for the community. Deletion of pelB resulted in a severe biofilm deficiency. Interestingly, this effect is strain-specific. Loss of Pel production in the laboratory strain PAO1 resulted in no difference in attachment or biofilm development; instead Psl proved to be the primary structural polysaccharide for biofilm maturity. Furthermore, we demonstrate that Pel plays a second role by enhancing resistance to aminoglycoside antibiotics. This protection occurs only in biofilm populations. We show that expression of the pel gene cluster and PelF protein levels are enhanced during biofilm growth compared to liquid cultures. Thus, we propose that Pel is capable of playing both a structural and a protective role in P. aeruginosa biofilms.

  18. Biochemical Aspects of Non-Starch Polysaccharides

    Directory of Open Access Journals (Sweden)

    Rodica Căpriţă

    2010-05-01

    Full Text Available Polysaccharides are macromolecules of monosaccharides linked by glycosidic bonds. Non-starch polysaccharides (NSP are principally non-α-glucan polysaccharides of the plant cell wall. They are a heterogeneous group of polysaccharides with varying degrees of water solubility, size, and structure. The water insoluble fiber fraction include cellulose, galactomannans, xylans, xyloglucans, and lignin, while the water-soluble fibers are the pectins, arabinogalactans, arabinoxylans, and β-(1,3(1,4-D-glucans (β-glucans. Knowledge of the chemical structure of NSP has permitted the development of enzyme technology to overcome their antinutritional effects. The physiological effects of NSP on the digestion and absorption of nutrients in human and monogastric animals have been attributed to their physicochemical properties: hydration properties, viscosity, cation exchange capacity and organic compound absorptive properties. This paper reviews and presents information on NSPs chemistry, physicochemical properties and physiological effects on the nutrient entrapment.

  19. NMR analysis of compositional heterogeneity in polysaccharides

    Science.gov (United States)

    Many copolysaccharides are compositionally heterogeneous, and the composition determined by the usual analytical or spectroscopic methods provides only an average value. For some polysaccharides, the NMR data contain copolymer sequence information, such as diad, triad, and tetrad sequence intensiti...

  20. Extraction Optimization of Polysaccharides from Pitaya Stems

    Institute of Scientific and Technical Information of China (English)

    HE Cong-fen; LI Peng; ZHAO Hua; SONG Li-ya; ZHU Jun; DONG Yin-mao

    2011-01-01

    [Objective] The aim was to describe the extraction of polysaccharides from pitaya stems.[Method] The hot water,enzyme-assisted and microwave-assisted methods were used,with the microwave-assisted extraction being deemed optimal by general evaluation.[Result] The main factors affecting the yield of polysaccharides in the microwave-assisted extraction,by order of magnitude,were as follows:timemicrowave powertemperature;additionally,optimal conditions included a 10 min extraction time,an 80 ℃ extraction temperature and a microwave setting of 200 W.Using these optimal conditions,the yield of PSPS(Polysaccharides from Pitaya Stems) was 1.42%.After purification,the yield of PSPS was 0.74%.[Conclusion] The PSPS was analyzed by IR,MALDI-TOF-MS and an element analysis technique.It was shown to be a polysaccharide mixture,and the molecular weight was between 3 900 and 4 300 Da.

  1. Electrospinning of polysaccharides for regenerative medicine.

    Science.gov (United States)

    Lee, Kuen Yong; Jeong, Lim; Kang, Yun Ok; Lee, Seung Jin; Park, Won Ho

    2009-10-05

    Electrospinning techniques enable the production of continuous fibers with dimensions on the scale of nanometers from a wide range of natural and synthetic polymers. The number of recent studies regarding electrospun polysaccharides and their derivatives, which are potentially useful for regenerative medicine, is increasing dramatically. However, difficulties regarding the processibility of the polysaccharides (e.g., poor solubility and high surface tension) have limited their application. In this review, we summarize the characteristics of various polysaccharides such as alginate, cellulose, chitin, chitosan, hyaluronic acid, starch, dextran, and heparin, which are either currently being used or have potential to be used for electrospinning. The recent progress of nanofiber matrices electrospun from polysaccharides and their biomedical applications in tissue engineering, wound dressings, drug delivery, and enzyme immobilization are discussed.

  2. Precipitation of low-temperature dolomite from an anaerobic microbial consortium: the role of methanogenic Archaea.

    Science.gov (United States)

    Kenward, P A; Goldstein, R H; González, L A; Roberts, J A

    2009-12-01

    Here we report precipitation of dolomite at low temperature (30 degrees C) mediated by a mixed anaerobic microbial consortium composed of dissimilatory iron-reducing bacteria (DIRB), fermenters, and methanogens. Initial solution geochemistry is controlled by DIRB, but after 90 days shifts to a system dominated by methanogens. In live experiments conditions are initially saturated with respect to dolomite (Omega(dol) = 19.40) and increase by two orders of magnitude (Omega(dol) = 2 330.77) only after the onset of methanogenesis, as judged by the increasing [CH(4)] and the detection of methanogenic micro-organisms. We identify ordered dolomite in live microcosms after 90 days via powder X-ray diffraction, while sterile controls precipitate only calcite. Scanning electron microscopy and transmitted electron microscopy demonstrate that the precipitated dolomite is closely associated with cell walls and putative extra-cellular polysaccharides. Headspace gas measurements and denaturing gradient gel electrophoresis confirm the presence of both autotrophic and acetoclastic methanogens and exclude the presence of DIRB and sulfate-reducing bacteria after dolomite begins forming. Furthermore, the absence of dolomite in the controls and prior to methanogenesis confirm that methanogenic Archaea are necessary for the low-temperature precipitation of dolomite under the experimental conditions tested.

  3. Soluble microbial products (SMPs release in activated sludge systems: a review

    Directory of Open Access Journals (Sweden)

    Azami Hamed

    2012-12-01

    Full Text Available Abstract This review discusses the characterization, production and implications of soluble microbial products (SMPs in biological wastewater treatment. The precise definition of SMPs is open to talk about, but is currently regarded as “the pool of organic compounds that are released into solution from substrate metabolism and biomass decay”'. Some of the SMPs have been identified as humic acids, polysaccharides, proteins, amino acids, antibiotics, extracellular enzymes and structural components of cells and products of energy metabolism. They adversely affect the kinetic activity, flocculating and settling properties of sludge. This review outlines some important findings with regard to biodegradability and treatability of SMPs and also the effect of process parameters on their production. As SMPs are produced during biological treatment process, their trace amounts normally remain in the effluent that defines the highest COD removal efficiency. Their presence in effluent represents a high potential risk of toxic by-product formation during chlorine disinfection. Studies have indicated that among all wastewater post-treatment processes, the adsorption by granular activated carbon combined with biologically induced degradation is the most effective method for removal of SMPs. However, it may be concludes that the knowledge regarding SMPs is still under progress and more work is required to fully understand their contribution to the treatment process.

  4. Microbial extracellular polymeric substances in marine biogeochemical processes

    Digital Repository Service at National Institute of Oceanography (India)

    Bhaskar, P.V.; Bhosle, N.B.

    for the fo ation and maintenance of microcolonies and oxygenfree con- ditions within the flocs that influence the spatial distribu- tion of different micro-organisms 68. Such niches in the micro-colonies become the hotspots for microbialaided organic... buried into the sediment. EPS in aggregation process In simple terms, aggregation is defined as a physical pro cess wherein micro particles collide and remain stuck to each other to form clumps of particles called aggrgates. These aggregates...

  5. Nanoparticle Facilitated Extracellular Electron Transfer in Microbial Fuel Cells

    Science.gov (United States)

    2014-10-13

    tances.19 Nanoscale materials and devices, such as metal /semi- conductor nanoparticles, nanowires, and carbon nanotubes, have been widely exploited...redox centers. In addition, dissimilatory intermembrane and intramembrane reduction of metals into nanoparticles has been well-documented29,30...different length scales. Biomineralized iron sulfide was employed for the current studies because of the importance of sulfur-mediated electron

  6. Extracellular DNA as matrix component in microbial biofilms

    DEFF Research Database (Denmark)

    Chiang, Wen-Chi; Tolker-Nielsen, Tim

    2010-01-01

    to various persistent infections in humans and animals, and to a variety of complications in industry, where solid–water interfaces occur. Knowledge about the molecular mechanisms involved in biofilm formation is necessary for creating strategies to control biofilms. Recent studies have shown...

  7. Extracellular matrix structure.

    Science.gov (United States)

    Theocharis, Achilleas D; Skandalis, Spyros S; Gialeli, Chrysostomi; Karamanos, Nikos K

    2016-02-01

    Extracellular matrix (ECM) is a non-cellular three-dimensional macromolecular network composed of collagens, proteoglycans/glycosaminoglycans, elastin, fibronectin, laminins, and several other glycoproteins. Matrix components bind each other as well as cell adhesion receptors forming a complex network into which cells reside in all tissues and organs. Cell surface receptors transduce signals into cells from ECM, which regulate diverse cellular functions, such as survival, growth, migration, and differentiation, and are vital for maintaining normal homeostasis. ECM is a highly dynamic structural network that continuously undergoes remodeling mediated by several matrix-degrading enzymes during normal and pathological conditions. Deregulation of ECM composition and structure is associated with the development and progression of several pathologic conditions. This article emphasizes in the complex ECM structure as to provide a better understanding of its dynamic structural and functional multipotency. Where relevant, the implication of the various families of ECM macromolecules in health and disease is also presented.

  8. Polysaccharides for the Delivery of Antitumor Drugs

    Directory of Open Access Journals (Sweden)

    Bianca Posocco

    2015-05-01

    Full Text Available Among the several delivery materials available so far, polysaccharides represent very attractive molecules as they can undergo a wide range of chemical modifications, are biocompatible, biodegradable, and have low immunogenic properties. Thus, polysaccharides can contribute to significantly overcome the limitation in the use of many types of drugs, including anti-cancer drugs. The use of conventional anti-cancer drugs is hampered by their high toxicity, mostly depending on the indiscriminate targeting of both cancer and normal cells. Additionally, for nucleic acid based drugs (NABDs, an emerging class of drugs with potential anti-cancer value, the practical use is problematic. This mostly depends on their fast degradation in biological fluids and the difficulties to cross cell membranes. Thus, for both classes of drugs, the development of optimal delivery materials is crucial. Here we discuss the possibility of using different kinds of polysaccharides, such as chitosan, hyaluronic acid, dextran, and pullulan, as smart drug delivery materials. We first describe the main features of polysaccharides, then a general overview about the aspects ruling drug release mechanisms and the pharmacokinetic are reported. Finally, notable examples of polysaccharide-based delivery of conventional anti-cancer drugs and NABDs are reported. Whereas additional research is required, the promising results obtained so far, fully justify further efforts, both in terms of economic support and investigations in the field of polysaccharides as drug delivery materials.

  9. Lytic polysaccharide monooxygenases: a crystallographer's view on a new class of biomass-degrading enzymes

    Directory of Open Access Journals (Sweden)

    Kristian E. H. Frandsen

    2016-11-01

    Full Text Available Lytic polysaccharide monooxygenases (LPMOs are a new class of microbial copper enzymes involved in the degradation of recalcitrant polysaccharides. They have only been discovered and characterized in the last 5–10 years and have stimulated strong interest both in biotechnology and in bioinorganic chemistry. In biotechnology, the hope is that these enzymes will finally help to make enzymatic biomass conversion, especially of lignocellulosic plant waste, economically attractive. Here, the role of LPMOs is likely to be in attacking bonds that are not accessible to other enzymes. LPMOs have attracted enormous interest since their discovery. The emphasis in this review is on the past and present contribution of crystallographic studies as a guide to functional understanding, with a final look towards the future.

  10. Polysaccharides serve as scaffold of biofilms formed by mucoid Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Yang, Liang; Hengzhuang, Wang; Wu, Hong

    2012-01-01

    Chronic lung infection by mucoid Pseudomonas aeruginosa is one of the major pathologic features in patients with cystic fibrosis. Mucoid P. aeruginosa is notorious for its biofilm forming capability and resistance to immune attacks. In this study, the roles of extracellular polymeric substances...... from biofilms formed by mucoid P. aeruginosa were investigated. Alginate is not an essential structure component for mucoid P. aeruginosa biofilms. Genetic studies revealed that Pel and Psl polysaccharides serve as essential scaffold and mediate macrocolony formation in mucoid P. aeruginosa biofilms...

  11. Escaping Underground Nets: Extracellular DNases Degrade Plant Extracellular Traps and Contribute to Virulence of the Plant Pathogenic Bacterium Ralstonia solanacearum.

    Science.gov (United States)

    Tran, Tuan Minh; MacIntyre, April; Hawes, Martha; Allen, Caitilyn

    2016-06-01

    Plant root border cells have been recently recognized as an important physical defense against soil-borne pathogens. Root border cells produce an extracellular matrix of protein, polysaccharide and DNA that functions like animal neutrophil extracellular traps to immobilize pathogens. Exposing pea root border cells to the root-infecting bacterial wilt pathogen Ralstonia solanacearum triggered release of DNA-containing extracellular traps in a flagellin-dependent manner. These traps rapidly immobilized the pathogen and killed some cells, but most of the entangled bacteria eventually escaped. The R. solanacearum genome encodes two putative extracellular DNases (exDNases) that are expressed during pathogenesis, suggesting that these exDNases contribute to bacterial virulence by enabling the bacterium to degrade and escape root border cell traps. We tested this hypothesis with R. solanacearum deletion mutants lacking one or both of these nucleases, named NucA and NucB. Functional studies with purified proteins revealed that NucA and NucB are non-specific endonucleases and that NucA is membrane-associated and cation-dependent. Single ΔnucA and ΔnucB mutants and the ΔnucA/B double mutant all had reduced virulence on wilt-susceptible tomato plants in a naturalistic soil-soak inoculation assay. The ΔnucA/B mutant was out-competed by the wild-type strain in planta and was less able to stunt root growth or colonize plant stems. Further, the double nuclease mutant could not escape from root border cells in vitro and was defective in attachment to pea roots. Taken together, these results demonstrate that extracellular DNases are novel virulence factors that help R. solanacearum successfully overcome plant defenses to infect plant roots and cause bacterial wilt disease.

  12. Escaping Underground Nets: Extracellular DNases Degrade Plant Extracellular Traps and Contribute to Virulence of the Plant Pathogenic Bacterium Ralstonia solanacearum.

    Directory of Open Access Journals (Sweden)

    Tuan Minh Tran

    2016-06-01

    Full Text Available Plant root border cells have been recently recognized as an important physical defense against soil-borne pathogens. Root border cells produce an extracellular matrix of protein, polysaccharide and DNA that functions like animal neutrophil extracellular traps to immobilize pathogens. Exposing pea root border cells to the root-infecting bacterial wilt pathogen Ralstonia solanacearum triggered release of DNA-containing extracellular traps in a flagellin-dependent manner. These traps rapidly immobilized the pathogen and killed some cells, but most of the entangled bacteria eventually escaped. The R. solanacearum genome encodes two putative extracellular DNases (exDNases that are expressed during pathogenesis, suggesting that these exDNases contribute to bacterial virulence by enabling the bacterium to degrade and escape root border cell traps. We tested this hypothesis with R. solanacearum deletion mutants lacking one or both of these nucleases, named NucA and NucB. Functional studies with purified proteins revealed that NucA and NucB are non-specific endonucleases and that NucA is membrane-associated and cation-dependent. Single ΔnucA and ΔnucB mutants and the ΔnucA/B double mutant all had reduced virulence on wilt-susceptible tomato plants in a naturalistic soil-soak inoculation assay. The ΔnucA/B mutant was out-competed by the wild-type strain in planta and was less able to stunt root growth or colonize plant stems. Further, the double nuclease mutant could not escape from root border cells in vitro and was defective in attachment to pea roots. Taken together, these results demonstrate that extracellular DNases are novel virulence factors that help R. solanacearum successfully overcome plant defenses to infect plant roots and cause bacterial wilt disease.

  13. Structural analysis of cell wall polysaccharides using PACE

    Energy Technology Data Exchange (ETDEWEB)

    Mortimer, Jennifer C. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Joint BioEnergy Institute

    2017-01-01

    The plant cell wall is composed of many complex polysaccharides. The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. Many techniques to characterize polysaccharide structure are complicated, requiring expensive equipment and specialized operators e.g. NMR, MALDI-MS. PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al. 2002). Whilst the method here describes xylan analysis, it can be applied (by use of the appropriate glycosyl hydrolase) to any cell wall polysaccharide.

  14. Recognition and degradation of plant cell wall polysaccharides by two human gut symbionts.

    Directory of Open Access Journals (Sweden)

    Eric C Martens

    2011-12-01

    Full Text Available Symbiotic bacteria inhabiting the human gut have evolved under intense pressure to utilize complex carbohydrates, primarily plant cell wall glycans in our diets. These polysaccharides are not digested by human enzymes, but are processed to absorbable short chain fatty acids by gut bacteria. The Bacteroidetes, one of two dominant bacterial phyla in the adult gut, possess broad glycan-degrading abilities. These species use a series of membrane protein complexes, termed Sus-like systems, for catabolism of many complex carbohydrates. However, the role of these systems in degrading the chemically diverse repertoire of plant cell wall glycans remains unknown. Here we show that two closely related human gut Bacteroides, B. thetaiotaomicron and B. ovatus, are capable of utilizing nearly all of the major plant and host glycans, including rhamnogalacturonan II, a highly complex polymer thought to be recalcitrant to microbial degradation. Transcriptional profiling and gene inactivation experiments revealed the identity and specificity of the polysaccharide utilization loci (PULs that encode individual Sus-like systems that target various plant polysaccharides. Comparative genomic analysis indicated that B. ovatus possesses several unique PULs that enable degradation of hemicellulosic polysaccharides, a phenotype absent from B. thetaiotaomicron. In contrast, the B. thetaiotaomicron genome has been shaped by increased numbers of PULs involved in metabolism of host mucin O-glycans, a phenotype that is undetectable in B. ovatus. Binding studies of the purified sensor domains of PUL-associated hybrid two-component systems in conjunction with transcriptional analyses demonstrate that complex oligosaccharides provide the regulatory cues that induce PUL activation and that each PUL is highly specific for a defined cell wall polymer. These results provide a view of how these species have diverged into different carbohydrate niches by evolving genes that target

  15. Metabolic requirements for neutrophil extracellular traps formation

    Science.gov (United States)

    Rodríguez-Espinosa, Oscar; Rojas-Espinosa, Oscar; Moreno-Altamirano, María Maximina Bertha; López-Villegas, Edgar Oliver; Sánchez-García, Francisco Javier

    2015-01-01

    As part of the innate immune response, neutrophils are at the forefront of defence against infection, resolution of inflammation and wound healing. They are the most abundant leucocytes in the peripheral blood, have a short lifespan and an estimated turnover of 1010 to 1011 cells per day. Neutrophils efficiently clear microbial infections by phagocytosis and by oxygen-dependent and oxygen-independent mechanisms. In 2004, a new neutrophil anti-microbial mechanism was described, the release of neutrophil extracellular traps (NETs) composed of DNA, histones and anti-microbial peptides. Several microorganisms, bacterial products, as well as pharmacological stimuli such as PMA, were shown to induce NETs. Neutrophils contain relatively few mitochondria, and derive most of their energy from glycolysis. In this scenario we aimed to analyse some of the metabolic requirements for NET formation. Here it is shown that NETs formation is strictly dependent on glucose and to a lesser extent on glutamine, that Glut-1, glucose uptake, and glycolysis rate increase upon PMA stimulation, and that NET formation is inhibited by the glycolysis inhibitor, 2-deoxy-glucose, and to a lesser extent by the ATP synthase inhibitor oligomycin. Moreover, when neutrophils were exposed to PMA in glucose-free medium for 3 hr, they lost their characteristic polymorphic nuclei but did not release NETs. However, if glucose (but not pyruvate) was added at this time, NET release took place within minutes, suggesting that NET formation could be metabolically divided into two phases; the first, independent from exogenous glucose (chromatin decondensation) and, the second (NET release), strictly dependent on exogenous glucose and glycolysis. PMID:25545227

  16. A comparative assessment of the potential of polysaccharide production and intracellular sugar composition within Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.:Fr.)P. Karst. (Aphyllophoromycetideae).

    Science.gov (United States)

    Stajić, Mirjana; Glamoclija, Jasmina; Maksimović, Vuk; Vukojević, Jelena; Simonić, Jasmina; Zervakis, George

    2011-01-01

    Ganoderma lucidum is a well-known medicinal mushroom species in which polysaccharides are one of the major sources of biological activity. The species was considered as a species-complex due to significant variations in morphological, biochemical, and genetic features among populations with a worldwide distribution. This fact was the basis for setting the aim of this research: to study intraspecific diversity in polysaccharide production and intracellular sugar composition among selected G. lucidum strains. The presence ofintraspecific diversity among 10 G. lucidum strains, from different areas worldwide, was noted. Values of produced mycelia biomass and intracellular polysaccharides were found in wide ranges (3.1 - 28.2 g L(-1) and 20.0 - 53.3 mg g(-1), respectively), while differences in extracellular polysaccharide amounts were minor (0.2 - 1.5 mg mL(-1)). The significant quantitative and qualitative differences in intracellular sugar composition were noted. Glucose was the predominant sugar in almost all strains except one (HAI 447), where sucrose was dominant. The potential of polysaccharide production and intracellular sugar composition could be one more taxonomic criterion for strain characterization within G. lucidum. The differences in intracellular sugar composition and proportions could be reflected in features of produced polysaccharides and also in their biological activities.

  17. Extracellular matrix structure governs invasion resistance in bacterial biofilms.

    Science.gov (United States)

    Nadell, Carey D; Drescher, Knut; Wingreen, Ned S; Bassler, Bonnie L

    2015-08-01

    Many bacteria are highly adapted for life in communities, or biofilms. A defining feature of biofilms is the production of extracellular matrix that binds cells together. The biofilm matrix provides numerous fitness benefits, including protection from environmental stresses and enhanced nutrient availability. Here we investigate defense against biofilm invasion using the model bacterium Vibrio cholerae. We demonstrate that immotile cells, including those identical to the biofilm resident strain, are completely excluded from entry into resident biofilms. Motile cells can colonize and grow on the biofilm exterior, but are readily removed by shear forces. Protection from invasion into the biofilm interior is mediated by the secreted protein RbmA, which binds mother-daughter cell pairs to each other and to polysaccharide components of the matrix. RbmA, and the invasion protection it confers, strongly localize to the cell lineages that produce it.

  18. A refined technique for extraction of extracellular matrices from bacterial biofilms and its applicability.

    Science.gov (United States)

    Chiba, Akio; Sugimoto, Shinya; Sato, Fumiya; Hori, Seiji; Mizunoe, Yoshimitsu

    2015-05-01

    Biofilm-forming bacteria embedded in polymeric extracellular matrices (ECMs) that consist of polysaccharides, proteins and/or extracellular DNAs (eDNAs) acquire high resistance to antimicrobial agents and host immune systems. To understand molecular mechanisms of biofilm formation and maintenance and to develop therapeutic countermeasures against chronic biofilm-associated infections, reliable methods to isolate ECMs are inevitable. In this study, we refined the ECM extraction method recently reported and evaluated its applicability. Using three Staphylococcus aureus biofilms in which proteins, polysaccharides or eDNAs are major contributors to their integrity, ECMs were extracted using salts and detergents. We found that extraction with 1.5 M sodium chloride (NaCl) could be optimum for not only ECM proteins but also polysaccharides and eDNAs. In addition, long-time incubation was not necessary for efficient ECM isolation. Lithium chloride (LiCl) was comparative to NaCl but is more expensive. In contrast to SDS, NaCl hardly caused leakage of intracellular proteins and did not affect viability of bacterial cells within biofilms. Furthermore, this method is applicable to other bacteria such as Gram-positive Staphylococcus epidermidis and Gram-negative Escherichia coli and Pseudomonas aeruginosa. Thus, this refined method is very simple, rapid, low cost and non-invasive and could be used for a broad range of applications.

  19. Immunomodulatory effects of a polysaccharide from Tamarindus indica.

    Science.gov (United States)

    Sreelekha, T T; Vijayakumar, T; Ankanthil, R; Vijayan, K K; Nair, M K

    1993-04-01

    A polysaccharide isolated and purified from Tamarindus indica shows immunomodulatory activities such as phagocytic enhancement, leukocyte migration inhibition and inhibition of cell proliferation. These properties suggest that this polysaccharide from T. indica may have some biological applications.

  20. Three-Dimensional Structural Aspects of Protein–Polysaccharide Interactions

    Directory of Open Access Journals (Sweden)

    Masamichi Nagae

    2014-03-01

    Full Text Available Linear polysaccharides are typically composed of repeating mono- or disaccharide units and are ubiquitous among living organisms. Polysaccharide diversity arises from chain-length variation, branching, and additional modifications. Structural diversity is associated with various physiological functions, which are often regulated by cognate polysaccharide-binding proteins. Proteins that interact with linear polysaccharides have been identified or developed, such as galectins and polysaccharide-specific antibodies, respectively. Currently, data is accumulating on the three-dimensional structure of polysaccharide-binding proteins. These proteins are classified into two types: exo-type and endo-type. The former group specifically interacts with the terminal units of polysaccharides, whereas the latter with internal units. In this review, we describe the structural aspects of exo-type and endo-type protein-polysaccharide interactions. Further, we discuss the structural basis for affinity and specificity enhancement in the face of inherently weak binding interactions.

  1. Ice nucleation activity of polysaccharides

    Science.gov (United States)

    Bichler, Magdalena; Felgitsch, Laura; Haeusler, Thomas; Seidl-Seiboth, Verena; Grothe, Hinrich

    2015-04-01

    Heterogeneous ice nucleation is an important process in the atmosphere. It shows direct impact on our climate by triggering ice cloud formation and therefore it has much influence on the radiation balance of our planet (Lohmann et al. 2002; Mishchenko et al. 1996). The process itself is not completely understood so far and many questions remain open. Different substances have been found to exhibit ice nucleation activity (INA). Due to their vast differences in chemistry and morphology it is difficult to predict what substance will make good ice nuclei and which will not. Hence simple model substances must be found and be tested regarding INA. Our work aims at gaining to a deeper understanding of heterogeneous ice nucleation. We intend to find some reference standards with defined chemistry, which may explain the mechanisms of heterogeneous ice nucleation. A particular focus lies on biological carbohydrates in regards to their INA. Biological carbohydrates are widely distributed in all kingdoms of life. Mostly they are specific for certain organisms and have well defined purposes, e.g. structural polysaccharides like chitin (in fungi and insects) and pectin (in plants), which has also water-binding properties. Since they are widely distributed throughout our biosphere and mostly safe to use for nutrition purposes, they are well studied and easily accessible, rendering them ideal candidates as proxies. In our experiments we examined various carbohydrates, like the already mentioned chitin and pectin, as well as their chemical modifications. Lohmann U.; A Glaciation Indirect Aerosol Effect Caused by Soot Aerosols; J. Geoph. Res.; Vol. 24 No.4; pp 11-1 - 11-4; 2002 Mishchenko M.I., Rossow W.B., Macke A., Lacis A. A.; Sensitivity of Cirrus Cloud Albedo, Bidirectional Reflectance and Optical Thickness Retrieval Accuracy to Ice Particle Shape, J. Geoph. Res.; Vol. 101, No D12; pp. 16,973 - 16,985; 1996

  2. Examination of some protective conditions on technological properties of irradiated food grade polysaccharides

    Energy Technology Data Exchange (ETDEWEB)

    Adeil Pietranera, M.S. E-mail: pietrane@cae.cnea.gov.ar; Narvaiz, P. E-mail: narvaiz@cae.cnea.gov.ar

    2001-02-01

    Corn and cassava starches, agar-agar and kappa carrageenan, polysaccharides used as food additives, were gamma irradiated with doses suitable for microbial decontamination. Starches were analysed in paste and pudding viscosity and paper chromatography; agar and carrageenan, in viscosity and compression tests on gels and custards. Afterwards, substances were added to verify whether they could prevent irradiation technological impairment. Results revealed significant differences between control and irradiated samples in most determinations. No added substance was able to protect the functional properties of these additives. (author)

  3. Elucidation of interaction mechanism between lacquer polysaccharides and proteins

    OpenAIRE

    Bai, Yuting

    2013-01-01

    Lacquer polysaccharides, which exist in the sap of Asian lacquer tree, are highly branched acidic polysaccharides with 1,3-β-galactan backbone and glucouronic acid terminals. Lacquer polysaccharides were reported to be antitumor active and blood coagulation promoting effective, which specific activities might be caused from electrostatic attraction between negative-charged carboxyl groups from the uronic acid terminal of polysaccharides, and positive-charged amino groups from target proteins....

  4. Capsular Polysaccharide Expression in Commensal Streptococcus Species

    DEFF Research Database (Denmark)

    Skov Sørensen, Uffe B; Yao, Kaihu; Yang, Yonghong

    2016-01-01

    Expression of a capsular polysaccharide is considered a hallmark of most invasive species of bacteria, including Streptococcus pneumoniae, in which the capsule is among the principal virulence factors and is the basis for successful vaccines. Consequently, it was previously assumed that capsule...... evolved by import of cps fragments from commensal Streptococcus species, resulting in a mosaic of genes of different origins. The demonstrated antigenic identity of at least eight of the numerous capsular polysaccharide structures expressed by commensal streptococci with recognized serotypes of S. pneumoniae...... of Streptococcus pneumoniae and is the basis for successful vaccines against infections caused by this important pathogen. Contrasting with previous assumptions, this study showed that expression of capsular polysaccharides by the same genetic mechanisms is a general property of closely related species...

  5. Influence of polysaccharides on wine protein aggregation.

    Science.gov (United States)

    Jaeckels, Nadine; Meier, Miriam; Dietrich, Helmut; Will, Frank; Decker, Heinz; Fronk, Petra

    2016-06-01

    Polysaccharides are the major high-molecular weight components of wines. In contrast, proteins occur only in small amounts in wine, but contribute to haze formation. The detailed mechanism of aggregation of these proteins, especially in combination with other wine components, remains unclear. This study demonstrates the different aggregation behavior between a buffer and a model wine system by dynamic light scattering. Arabinogalactan-protein, for example, shows an increased aggregation in the model wine system, while in the buffer system a reducing effect is observed. Thus, we could show the importance to examine the behavior of wine additives under conditions close to reality, instead of simpler buffer systems. Additional experiments on melting points of wine proteins reveal that only some isoforms of thaumatin-like proteins and chitinases are involved in haze formation. We can confirm interactions between polysaccharides and proteins, but none of these polysaccharides is able to prevent haze in wine.

  6. Depolymerization of sulfated polysaccharides under hydrothermal conditions.

    Science.gov (United States)

    Morimoto, Minoru; Takatori, Masaki; Hayashi, Tetsuya; Mori, Daiki; Takashima, Osamu; Yoshida, Shinichi; Sato, Kimihiko; Kawamoto, Hitoshi; Tamura, Jun-ichi; Izawa, Hironori; Ifuku, Shinsuke; Saimoto, Hiroyuki

    2014-01-30

    Fucoidan and chondroitin sulfate, which are well known sulfated polysaccharides, were depolymerized under hydrothermal conditions (120-180°C, 5-60min) as a method for the preparation of sulfated polysaccharides with controlled molecular weights. Fucoidan was easily depolymerized, and the change of the molecular weight values depended on the reaction temperature and time. The degree of sulfation and IR spectra of the depolymerized fucoidan did not change compared with those of untreated fucoidan at reaction temperatures below 140°C. However, fucoidan was partially degraded during depolymerization above 160°C. Nearly the same depolymerization was observed for chondroitin sulfate. These results indicate that hydrothermal treatment is applicable for the depolymerization of sulfated polysaccharides, and that low molecular weight products without desulfation and deformation of the initial glycan structures can be obtained under mild hydrothermal conditions.

  7. Capsular Polysaccharide Expression in Commensal Streptococcus Species

    DEFF Research Database (Denmark)

    Skov Sørensen, Uffe B; Yao, Kaihu; Yang, Yonghong

    2016-01-01

    biosynthesis (cps) loci in all strains tested. Truncated cps loci were detected in three strains of S. pseudopneumoniae, in 26% of S. mitis strains, and in a single S. oralis strain. The level of sequence identities of cps locus genes confirmed that the structural polymorphism of capsular polysaccharides in S....... pneumoniae evolved by import of cps fragments from commensal Streptococcus species, resulting in a mosaic of genes of different origins. The demonstrated antigenic identity of at least eight of the numerous capsular polysaccharide structures expressed by commensal streptococci with recognized serotypes of S...... of streptococci that form a significant part of our commensal microbiota. The demonstrated antigenic identity of many capsular polysaccharides expressed by commensal streptococci and S. pneumoniae raises important questions concerning the consequences for vaccination and host-parasite relationships both...

  8. Semi-synthesis of unusual chondroitin sulfate polysaccharides containing GlcA(3-O-sulfate) or GlcA(2,3-di-O-sulfate) units.

    Science.gov (United States)

    Bedini, Emiliano; De Castro, Cristina; De Rosa, Mario; Di Nola, Annalida; Restaino, Odile F; Schiraldi, Chiara; Parrilli, Michelangelo

    2012-02-13

    The extraction from natural sources of Chondroitin sulfate (CS), a polysaccharide used for management of osteoarthritis, leads to very complex mixtures. The synthesis of CS by chemical modification of other polysaccharides has seldom been reported due to the intrinsic complexity that arises from fine chemical modifications of the polysaccharide structure. In view of the growing interest in expanding the application of CS to pharmacological fields other than osteoarthritis treatment, we launched a program to find new sources of known or even unprecedented CS polysaccharides. As part of this program, we report herein on an investigation of the use of a cyclic orthoester group to selectively protect the 4,6-diol of N-acetyl-galactosamine residues in chondroitin (obtained from a microbial source), thereby facilitating its transformation into CSs. In particular, three CS polysaccharides were obtained and demonstrated to possess rare or hitherto unprecedented sulfation patterns by 2D NMR spectroscopy characterization. Two of them contained disaccharide subunits characterized by glucuronic acid residues selectively sulfated at position 3 (GlcA(3S)), the biological functions of which are known but have yet to be fully investigated. This first semi-synthetic access to GlcA(3S)-containing CS could greatly expedite such studies, since it can easily furnish considerable amounts of these polysaccharides, which are usually isolated with difficulty and in very low quantity from natural sources.

  9. Anionic Polyacrylamide (PAM) and Extracellular Polysaccharides (EPS) effects on flocculation and aggregate stability of soil

    Science.gov (United States)

    Albalasmeh, A. A.; Gharaibeh, M. A.; Ghezzehei, T. A.

    2015-12-01

    Soil structure influences many soil properties including aeration, water retention, drainage, bulk density, and resistance to erosion and indirectly influences most biological and chemical processes that occur in and around soil. A significant amount of literature showed that PAM plays an important role to control erosion. However, researchers are looking for more natural alternative for PAM. This study evaluated two anionic polymers including low and high molecular weight (MW), root exudates and bacterial exudates. We evaluated their influence on the rate and efficacy of colloid flocculation and the percent of water stable aggregates. We found that PAM was more effective than EPS in flocculating the colloids and all polymers increased the percent of stable soil aggregates although the PAM was more effective. These data suggest that the EPS would be less effective than PAM for reducing water erosion owing to its lesser flocculation and aggregate stabilizing potential.

  10. Screening and characterization of extracellular polysaccharides produced by Leuconostoc kimchii isolated from traditional fermented pulque beverage.

    Science.gov (United States)

    Torres-Rodríguez, Ingrid; Rodríguez-Alegría, María Elena; Miranda-Molina, Alfonso; Giles-Gómez, Martha; Conca Morales, Rodrigo; López-Munguía, Agustín; Bolívar, Francisco; Escalante, Adelfo

    2014-01-01

    We report the screening and characterization of EPS produced by LAB identified as Leuconostoc kimchii isolated from pulque, a traditional Mexican fermented, non-distilled alcoholic beverage produced by the fermentation of the sap extracted from several (Agave) maguey species. EPS-producing LAB constitutes an abundant bacterial group relative to total LAB present in sap and during fermentation, however, only two EPS-producing colony phenotypes (EPSA and EPSB, respectively) were detected and isolated concluding that despite the high number of polymer-producing LAB their phenotypic diversity is low. Scanning electron microcopy analysis during EPS-producing conditions revealed that both types of EPS form a uniform porous structure surrounding the bacterial cells. The structural characterization of the soluble and cell-associated EPS fractions of each polymer by enzymatic and acid hydrolysis, as by 1D- and 2D-NMR, showed that polymers produced by the soluble and cell-associated fractions of EPSA strain are dextrans consisting of a linear backbone of linked α-(1→6) Glcp in the main chain with α-(1→2) and α-(1→3)-linked branches. The polymer produced by the soluble fraction of EPSB strain was identified as a class 1 dextran with a linear backbone containing consecutive α-(1→6)-linked D-glucopyranosyl units with few α-(1→3)-linked branches, whereas the cell-associated EPS is a polymer mixture consisting of a levan composed of linear chains of (2→6)-linked β-D-fructofuranosyl residues with β-(2→6) connections, and a class 1 dextran. According to our knowledge this is the first report of dextrans and a levan including their structural characterization produced by L. kimchii isolated from a traditional fermented source.

  11. Endurance of Australian dry-savannah cyanobacteria regulated by extra-cellular polysaccharides - EPS and environment

    Science.gov (United States)

    Williams, W.; Budel, B.; Reichenberger, H.; Rose, N.

    2012-04-01

    Cyanobacterial crusts are an important driver of ecosystem function throughout Queensland's dry savannah. Annually there is very little rainfall during the winter-dry season. In the summer-wet season build-up early storms precede its onset; days are low in humidity with high ambient (>40°C) and soil surface temperatures (60-74°C). In the wet season monsoon rains and tropical storms result in vast flooded plains and ephemeral wetlands, leaving the ground saturated for several weeks. At Boodjamulla National Park (NW Qld), cyanobacterial crusts were sampled during the dry season, after 125 days without rain. An Imaging PAM (Walz) was used to determine the resurrection and quantum yield of Photosystem II (PSII). The crusts were periodically watered for ten days and multiple PAM measurements were made on a daily basis. PSII in cyanobacteria showed no signs of resurrection; however new Nostoc colonies emerged on the eighth day. Microscopic examination revealed other cyanobacteria remained in a desiccated state and EPS seemed hydrophobic. In the following dry season, crust samples were preserved at 40°C at low humidity. During the wet season these samples were reintroduced into their natural environment of high humidity and subject to periodic rains. The resurrection of PSII commenced within two hours of the first rainfall and was fully functional within 24 hours at which time existing cyanobacterial cells rapidly re-hydrated and EPS exhibited hydrophilicity. These are the first field studies demonstrating the environmental conditions controlling the function of cyanobacterial EPS and the resurrection of PSII. Mass EPS production occurs several times throughout the wet season. Eventually, as the humidity drops, temperatures remain high and sunny conditions prevail the EPS hardens and dries forming thick hydrophobic polymeric surfaces. As temperatures rise and fall, crusts crack and curl, then start to disintegrate after the first rains of the wet season before new crusts start to grow.

  12. [Component analysis on polysaccharides in exocarp of Ginkgo biloba].

    Science.gov (United States)

    Song, G; Xu, A; Chen, H; Wang, X

    1997-09-01

    This paper reports the content and component analysis on polysaccharides in exocarp of Ginkgo biloba. The results show that the content of total saccharides is 89.7%; content of polysaccharides is 84.6%; content of reductic saccharides is 5.1%; the polysaccharides are composed of glucose, fructose, galactose and rhamnose.

  13. Extracellular vesicles for drug delivery

    NARCIS (Netherlands)

    Vader, Pieter; Mol, Emma A; Pasterkamp, Gerard; Schiffelers, Raymond M

    Extracellular vesicles (EVs) are cell-derived membrane vesicles, and represent an endogenous mechanism for intercellular communication. Since the discovery that EVs are capable of functionally transferring biological information, the potential use of EVs as drug delivery vehicles has gained

  14. Studies on Sulfation of Lycium barbarum Polysaccharides

    Institute of Scientific and Technical Information of China (English)

    YI,Jian-Ping; YAN,Hong; ZHONG,Ru-Gang

    2004-01-01

    @@ Polysaccharides can anti-virus, such as human immunodeficiency virus (HIV-1),[1] herpes simplex virus (HSV-1,HSV-2) and cytomegalovirus. Some of them are sulfates, e.g. dextran sulfate, heparin, sulfonation of chitosan and sulfated derivatives of Lentinan. Our results showed that sulfated derivatives of Lycium barbarum polysaccharides (LBP)have anti-HIV activity. Because the anti-HIV activity of LBP was deeply dependent on the molecular weight, the sulfation pattern and glycosidic branches besides degree of sulfation (DS), so we emphasized our work on the factors of DS.

  15. Galactomannan: a versatile biodegradable seed polysaccharide.

    Science.gov (United States)

    Prajapati, Vipul D; Jani, Girish K; Moradiya, Naresh G; Randeria, Narayan P; Nagar, Bhanu J; Naikwadi, Nikhil N; Variya, Bhavesh C

    2013-09-01

    Polysaccharides have been finding, in the last decades, very interesting and useful applications in the biomedical and, specifically, in the biopharmaceutical field. Galactomannans are a group of storage polysaccharides from various plant seeds that reserve energy for germination in the endosperm. There are four major sources of seed galactomannans: locust bean (Ceratonia siliqua), guar (Cyamopsis tetragonoloba), tara (Caesalpinia spinosa Kuntze), and fenugreek (Trigonella foenum-graecum L.). Through keen references of reported literature on galactomannans, in this review, we have described occurrence of various galactomannans, its physicochemical properties, characterization, applications, and overview of some major galactomannans.

  16. Influence of substrate and microbial interaction on efficiency of rumen microbial growth.

    Science.gov (United States)

    Demeyer, D; Van Nevel, C

    1986-01-01

    Microbial N produced in the rumen and flowing to the duodenum (Ni) is related to the total amount of OM fermented or apparently digested in the rumen (OMf). This relationship, best expressed as microbial N yield (gNi/kgOMf), is affected mainly by the physical and chemical properties of feed carbohydrates and the amounts ingested. These factors influence yields at three levels of increasing complexity: Bacterial fermentation within one compartment following the continuous culture model. Fermentation pattern as such does not seem to affect yields. High fermentation rates are associated with lactate production, low methane production and transient polysaccharide synthesis. These effects induce acidification and lower yields, partly compensated by faster growth. Protozoal action, determined by the presence of sequestration spaces provided mainly by roughage diets. The presence of protozoa depresses microbial N yield but allows more complete fibre digestion. Compartmentation and differential passage. With roughage diets, optimal microbial N yield seems to require well developed microbial compartmentation, involving a large proportion of microbes in a large-particle pool with a slow turnover, balanced by a small proportion in liquid, small-particle pools with a fast turnover. Such a situation is associated with long roughage feeding. It is hypothesized that microbial N yields in the rumen may vary between two extremes which are associated with the feeding of long roughage on the one hand or with concentrate (starch) feeding on the other.

  17. Extensive microbial and functional diversity within the chicken cecal microbiome.

    Directory of Open Access Journals (Sweden)

    Martin J Sergeant

    Full Text Available Chickens are major source of food and protein worldwide. Feed conversion and the health of chickens relies on the largely unexplored complex microbial community that inhabits the chicken gut, including the ceca. We have carried out deep microbial community profiling of the microbiota in twenty cecal samples via 16S rRNA gene sequences and an in-depth metagenomics analysis of a single cecal microbiota. We recovered 699 phylotypes, over half of which appear to represent previously unknown species. We obtained 648,251 environmental gene tags (EGTs, the majority of which represent new species. These were binned into over two-dozen draft genomes, which included Campylobacter jejuni and Helicobacter pullorum. We found numerous polysaccharide- and oligosaccharide-degrading enzymes encoding within the metagenome, some of which appeared to be part of polysaccharide utilization systems with genetic evidence for the co-ordination of polysaccharide degradation with sugar transport and utilization. The cecal metagenome encodes several fermentation pathways leading to the production of short-chain fatty acids, including some with novel features. We found a dozen uptake hydrogenases encoded in the metagenome and speculate that these provide major hydrogen sinks within this microbial community and might explain the high abundance of several genera within this microbiome, including Campylobacter, Helicobacter and Megamonas.

  18. Polysaccharide charge density regulating protein adsorption to air/water interfaces by protein/polysaccharide complex formation

    NARCIS (Netherlands)

    Ganzevles, R.A.; Kosters, H.; Vliet, T. van; Stuart, M.A.C.; Jongh, H.H.J. de

    2007-01-01

    Because the formation of protein/polysaccharide complexes is dominated by electrostatic interaction, polysaccharide charge density is expected to play a major role in the adsorption behavior of the complexes. In this study, pullulan (a non-charged polysaccharide) carboxylated to four different

  19. Influences of environmental factors on bacterial extracellular polymeric substances production in porous media

    Science.gov (United States)

    Xia, Lu; Zheng, Xilai; Shao, Haibing; Xin, Jia; Peng, Tao

    2014-11-01

    Bioclogging of natural porous media occurs frequently under a wide range of conditions. It may influence the performance of permeable reactive barrier and constructed wetland. It is also one of the factors that determine the effect of artificial groundwater recharge and in situ bioremediation process. In this study, a series of percolation column experiments were conducted to simulate bioclogging process in porous media. The predominant bacteria in porous media which induced clogging were identified to be Methylobacterium, Janthinobacterium, Yersinia, Staphylococcus and Acidovorax, most of which had been shown to effectively produce viscous extracellular polymeric substances (EPS). The column in which EPS production was maximized also coincided with the largest reduction in saturated hydraulic conductivity of porous media. In addition, carbon concentration was the most significant factor to affect polysaccharide, protein and EPS secretion, followed by phosphorus concentration and temperature. The coupled effect of carbon and phosphorus concentration was also very important to stimulate polysaccharide and EPS production.

  20. Production of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt. :Fr.) P. Karst. (higher Basidiomycetes), biomass and polysaccharides by solid state cultivation.

    Science.gov (United States)

    Berovic, Marin; Habijanic, Jozica; Boh, Bojana; Wraber, Branka; Petravic-Tominac, Vlatka

    2012-01-01

    Solid state cultivation of Ganoderma lucidum biomass, strain BFWS Gal 4, originally isolated from the Slovenian forest, was studied in a horizontal stirred tank reactor. Periodic mixing of N = 80 rpm, 2 min/day was used. Production of fungal polysaccharides and fungal biomass on solid substrate based on beech sawdust, olive oil, and mineral salts was studied. Optimal moisture of the solid matrix was in the range of 80% to 74%. When the moisture content dropped below 57%, the growth of the mycelium and polysaccharide production stopped, but it revived when wet air was applied in further processing. Final concentration of biomass was 0.68 mg/g of solid substrate, while proportions of extracellular and intracellular polysaccharides were 4.5 mg/g and 1.05 mg/g, respectively.

  1. Effects of low-level laser therapy combined with toluidine blue on polysaccharides and biofilm of Streptococcus mutans.

    Science.gov (United States)

    de Sousa Farias, S S; Nemezio, M A; Corona, S A M; Aires, C P; Borsatto, M C

    2016-07-01

    The aim of this study was to evaluate the effect of a low-level laser therapy in combination with toluidine blue on polysaccharides and biofilm of Streptococcus mutans. S. mutans biofilms were formed on acrylic resin blocks. These biofilms were exposed eight times/day to 10 % sucrose, and two times/day, they were subjected to one of the following treatments: G1, 0.9 % NaCl as a negative control; G2, 0.12 % chlorhexidine digluconate (CHX) as a positive antibacterial control; and G3 and G4 antimicrobial photodynamic therapy (aPDT) combined with toluidine blue using dosages of 320 and 640 J/cm(2), respectively. The experiment was performed in triplicate. The biofilm formed on each block was collected for determination of the viable bacteria and concentration of insoluble extracellular polysaccharides (IEPS) and intracellular polysaccharides (IPS). CHX and aPDT treatments were able to inhibit bacterial growth in comparison with negative control (p  0.05). The results suggest that low-level laser therapy presents effects on biofilm bacteria viability and in polysaccharides concentration.

  2. Polysaccharides from Ganoderma lucidum Promote Cognitive Function and Neural Progenitor Proliferation in Mouse Model of Alzheimer's Disease

    Directory of Open Access Journals (Sweden)

    Shichao Huang

    2017-01-01

    Full Text Available Promoting neurogenesis is a promising strategy for the treatment of cognition impairment associated with Alzheimer's disease (AD. Ganoderma lucidum is a revered medicinal mushroom for health-promoting benefits in the Orient. Here, we found that oral administration of the polysaccharides and water extract from G. lucidum promoted neural progenitor cell (NPC proliferation to enhance neurogenesis and alleviated cognitive deficits in transgenic AD mice. G. lucidum polysaccharides (GLP also promoted self-renewal of NPC in cell culture. Further mechanistic study revealed that GLP potentiated activation of fibroblast growth factor receptor 1 (FGFR1 and downstream extracellular signal-regulated kinase (ERK and AKT cascades. Consistently, inhibition of FGFR1 effectively blocked the GLP-promoted NPC proliferation and activation of the downstream cascades. Our findings suggest that GLP could serve as a regenerative therapeutic agent for the treatment of cognitive decline associated with neurodegenerative diseases.•G. lucidum polysaccharides (GLP improve cognition in transgenic AD mice•GLP promote neural progenitor proliferation and self-renewal to enhance neurogenesis•GLP potentiate FGFR signalingG. lucidum is a revered medicinal herb for promoting health. Pei and colleagues found that the polysaccharides from G. lucidum (GLP promoted neural progenitor proliferation to enhance neurogenesis and ameliorated cognition deficits in transgenic Alzheimer's disease model mice. These findings suggest that GLP could serve as a regenerative therapeutic agent for the treatment of cognitive decline associated with neurodegenerative diseases.

  3. Lentinula edodes-derived polysaccharide alters the spatial structure of gut microbiota in mice.

    Directory of Open Access Journals (Sweden)

    Xiaofei Xu

    Full Text Available Lentinula edodes-derived polysaccharides possess many therapeutic characteristics, including anti-tumor and immuno-modulation. The gut microbes play a critical role in modulation of immune function. However, the impact of Lentinula edodes-derived polysaccharides on the gut microbes have not yet been explored. In this study, high-throughput pyrosequencing technique was employed to investigate the effects of a new heteropolysaccharide L2 from Lentinula edodes on microbiota diversity and composition of small intestine, cecum, colon and distal end of colon (feces in mice. The results demonstrated that along mouse intestine the microbiota exhibit distinctly different space distribution. L2 treatment reduced the diversity and evenness of gut microbiota along the intestine, especially in the cecum and colon. In the fecal microbial communities, the decrease of Bacteroidetes by significantly increasing Proteobacteria were observed, which were characterized by the increased Helicobacteraceae and reduced S24-7 at family level. Some OTUs, corresponding to Bacteroides acidifaciens, Alistipes and Helicobacter suncus, were found to be significantly increased in L2 treated-mice. In particular, 4 phyla Chloroflexi, Gemmatimonadetes, Nitrospirae and Planctomycetes are exclusively present in L2-treated mice. This is helpful for further demonstrating healthy action mechanism of Lentinula edodes-derived polysaccharide L2.

  4. Effect of proteins, polysaccharides, and particle sizes on sludge dewaterability

    Institute of Scientific and Technical Information of China (English)

    SHAO Liming; HE Peipei; YU Guanghui; HE Pinjing

    2009-01-01

    Four batch experiments of hydrolysis and acidification were carried out to investigate the distributions of proteins (PN) and polysaccharides (PS) in the sludge, the PN/PS ratio, the particle sizes, and their relationship with sludge dewaterability (as determined by capillary suction time, CST). The sludge flocs were stratified through centrifugation- and ultrasound-based method into four layers: (1) slime, (2) loosely bound extracellular polymeric substances (LB-EPS), (3) tightly bound EPS (TB-EPS), and (4) pellet. The results showed that PN was mainly partitioned in the pellet (80.7%) and TB-EPS (9.6%) layers, while PS distributed evenly in the four layers. During hydrolysis and acidification, PN was transferred from the pellet and TB-EPS layers to the slime layer, but PS had no significant transfer trends. The mean particle sizes of the sludge flocs decreased with hydrolysis and acidification. The pH had a more significant influence on the dewaterability of sludge flocs than temperature. Sludge dewaterability during hydrolysis and acidification processes greatly deteriorated from 9.7 s at raw sludge to 340--450 s under alkaline conditions. However, it was just slightly increased under acidic conditions. Further investigation suggested that CST was affected by soluble PN, soluble PN/PS, and particle sizes of sludge flocs, but was affected slightly by total PN, PS, or PN/PS in the whole sludge flocs and other layers (except slime).

  5. Analysis of Lignin-Polysaccharide Complexes Formed during Grass Lignin Degradation by Cultures of Pleurotus Species.

    Science.gov (United States)

    Gutierrez, A; Bocchini, P; Galletti, G C; Martinez, A T

    1996-06-01

    A brown material, precipitable with ethanol, was formed during wheat straw and lignin degradation by liquid cultures of different species of Pleurotus. Fourier transform infrared spectroscopy and cross-polarization and magic-angle-spinning (sup13)C nuclear magnetic resonance spectroscopy showed that most of the precipitable material was formed from exopolysaccharide secreted by the fungus but it also contained an aromatic fraction. The results of acid hydrolysis, methylation analysis, and Smith degradation indicated that the major exopolysaccharide produced by these fungi is a (1(symbl)3)-(beta)-glucan branched at C-6 every two or three residues along the main chain. The presence of lignin or straw in the culture medium had little effect on the composition and structure of the extracellular polysaccharide. Cross-polarization and magic-angle-spinning (sup13)C nuclear magnetic resonance spectroscopy provided an estimation of the aromatic content of the lignin-polysaccharide complexes, assigning 20% of the total (sup13)C signal in the material recovered from cultures of Pleurotus eryngii in lignin medium to aromatic carbon. Analytical pyrolysis indicated that the aromatic fractions of the lignin-polysaccharide complexes were derived from lignin, since products characteristic of pyrolytic breakdown of H (p-hydroxyphenylpropane), G (guaiacylpropane), and S (syringylpropane) lignin units were identified. These complexes cannot be fractionated by treatment with polyvinylpyrrolidone or extraction with lignin solvents, suggesting that the two polymers were chemically linked. Moreover, differences in composition with respect to the original lignin indicated that this macromolecule was modified by the fungi during the process of formation of the lignin-polysaccharide complexes.

  6. Microbial xanthophylls.

    Science.gov (United States)

    Bhosale, Prakash; Bernstein, Paul S

    2005-09-01

    Xanthophylls are oxygenated carotenoids abundant in the human food supply. Lutein, zeaxanthin, and cryptoxanthin are major xanthophyll carotenoids in human plasma. The consumption of these xanthophylls is directly associated with reduction in the risk of cancers, cardiovascular disease, age-related macular degeneration, and cataract formation. Canthaxanthin and astaxanthin also have considerable importance in aquaculture for salmonid and crustacean pigmentation, and are of commercial interest for the pharmaceutical and food industries. Chemical synthesis is a major source for the heavy demand of xanthophylls in the consumer market; however, microbial producers also have potential as commercial sources. In this review, we discuss the biosynthesis, commercial utility, and major microbial sources of xanthophylls. We also present a critical review of current research and technologies involved in promoting microbes as potential commercial sources for mass production.

  7. Improved coupling of bacterial polysaccharides to macromolecules and solid supports

    DEFF Research Database (Denmark)

    2013-01-01

    The invention relates to a method of producing a polysaccharide-carrier conjugate comprising coupling a polysaccharide to a carrier, said polysaccharide comprising at least one monosaccharide unit comprising a keto-carboxy group according to the formula -C(=O)COOR, where R is either hydrogen or C1......-alkoxyamine group of the carrier with a keto-carboxy group of said polysaccharide to form a covalent amide bond between the carrier and the polysaccharide. Another aspect of the present invention relates to a compound or solid surface obtained when performing the method of the present invention. A third aspect...

  8. SORPTION PROPERTIES OF PLANT POLYSACCHARIDE COMPLEXES

    Directory of Open Access Journals (Sweden)

    L. E. Glagoleva

    2012-01-01

    Full Text Available The article presents information on the laws of the sorption of water to grow-negative polysaccharide complexes of the pumpkin and briar, deter-mined the rate constant of swelling as a function of temperature and pH, the maximum degree of swelling and limit the time to achieve it.

  9. Fucoidans - sulfated polysaccharides of brown algae

    Energy Technology Data Exchange (ETDEWEB)

    Usov, Anatolii I; Bilan, M I [N.D.Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow (Russian Federation)

    2009-08-31

    The methods of isolation of fucoidans and determination of their chemical structures are reviewed. The fucoidans represent sulfated polysaccharides of brown algae, the composition of which varies from simple fucan sulfates to complex heteropolysaccharides. The currently known structures of such biopolymers are presented. A variety of the biological activities of fucoidans is briefly summarised.

  10. Fucoidans — sulfated polysaccharides of brown algae

    Science.gov (United States)

    Usov, Anatolii I.; Bilan, M. I.

    2009-08-01

    The methods of isolation of fucoidans and determination of their chemical structures are reviewed. The fucoidans represent sulfated polysaccharides of brown algae, the composition of which varies from simple fucan sulfates to complex heteropolysaccharides. The currently known structures of such biopolymers are presented. A variety of the biological activities of fucoidans is briefly summarised.

  11. Polysaccharides from Polygonatum odoratum strengthen antioxidant ...

    African Journals Online (AJOL)

    . Purpose: To investigate the potential effects of polysaccharides isolated from ... groups were treated with different doses (150, 300, and 600 mg/kg) of PPO, ..... anti-fatigue properties. .... Zhao XN, Liang JL, Chen HB, Liang YE, Guo HZ, Su ZR,.

  12. Sustainability, polysaccharide science, and bio-economy

    NARCIS (Netherlands)

    Bos, Ten R.; Dam, Van J.E.G.

    2013-01-01

    At the opening of the 2nd EPNOE conference the role and responsibility of polysaccharide scientists was reflected upon and placed in the context of actual global issues like the transition process towards “sustainable bio-economy”. Difficulties in the chain of communication between the different

  13. Enzymatic production of polysaccharides from gum tragacanth

    DEFF Research Database (Denmark)

    2014-01-01

    Plant polysaccharides, relating to the field of natural probiotic components, can comprise structures similar to human milk oligosaccharides. A method for enzymatic hydrolysis of gum tragacanth from the bush-like legumes of the genus Astragalus, using a combination of pectin hydrolases...

  14. Polysaccharide coating of human corneal endothelium

    DEFF Research Database (Denmark)

    Schroder, H D; Sperling, S

    1977-01-01

    Electron microscopy revealed the presence of a 600-1500 A thick layer of polysaccharide on the surface of human corneal endothelial cells. The surface layer was visualized by combined fixation and staining in a mixture of ruthenium red and osmium tetroxide. The coating material was stable...

  15. Immunomodulatory activities of five clinically used Chinese herbal polysaccharides

    Directory of Open Access Journals (Sweden)

    Aiping Lu

    2012-02-01

    Full Text Available Polysaccharide is a natural macromolecular compound with complex, important and multifaceted biological activities. Some of polysaccharides have been marketed in China as drugs or healthy products. More studies confirm that the active ingredient of many traditional Chinese medicine exist in the form of polysaccharides. They play a role in disease therapy by activating immune cells and the complement system; regulating the cytokines expression; promoting the production of antibodies; inhibiting tumor cell proliferation and inducing tumor cell apoptosis; inhibiting virus entering cells and replication; increasing activity of antioxidant enzyme; scavenging free radicals; and inhibiting lipid peroxidation. In this review, we focus on the immunomodulatory effects and its possible mechanism of polysaccharides from Chinese herbal polysaccharides products, including Lentinan, Astragalus polysaccharide, Polyporus polysaccharide and Achyranthes bidentata polysaccharide. The immunomodulatory activities of polysaccharides were categorized in the paper into general immunoregulatory activity, anti-tumor, anti-infections, anti-inflammatory, anti-oxidative, anti-mutagenic and radioprotective, anti-complementary, anti-adhesive, and anti-allergy since all the activities are related to modulate immune responses by the polysaccharides. Also the challenges in the research of polysaccharides will be discussed. [J Exp Integr Med 2012; 2(1.000: 15-27

  16. Microscopic monitoring of extracellular pH in dental biofilms

    DEFF Research Database (Denmark)

    Schlafer, Sebastian; Garcia, Javier; Greve, Matilde

    pH in dental biofilm is a key virulence factor for the development of caries lesions. The complex three-dimensional architecture of dental biofilms leads to steep gradients of nutrients and metabolites, including organic acids, across the biofilm. For decades, measuring pH in dental biofilm has...... been limited to monitoring bulk pH with electrodes. Although pH microelectrodes with a better spatial resolution have been developed, they do not permit to monitor horizontal pH gradients in real-time. Quantitative fluorescent microscopic techniques, such as fluorescence lifetime imaging or pH...... ratiometry, can be employed to map the pH landscape in dental biofilm with more detail. However, when pH sensitive fluorescent probes are used to visualize pH in biofilms, it is crucial to differentiate between extracellular and intracellular pH. Intracellular microbial pH and pH in the extracellular matrix...

  17. Differential role of eDNA, proteins, and polysaccharides in cell-cell and cell-substrate adhesion by three Staphylococcus species

    DEFF Research Database (Denmark)

    Meyer, Rikke Louise; Okshevsky, Mira Ursula; Zeng, Guanghong

    on abiotic surfaces. We quantified initial adhesion, cell aggregation, and single-cell adhesion forces of Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus xylosus in the presence and absence of DNase, dispersin, or subtilisin, which cleave extracellular DNA, polysaccharides and proteins...... valuable for designing new approaches to biofilm prevention. In this study, we combine microfluidic flow-cell studies with single-cell analyses to understand how polysaccharides, extracellular DNA (eDNA), and proteins contribute individually and in concert to mediate bacterial adhesion and aggregation...... eDNA and proteins are the most important adhesins for initiating S. aureus biofilms. S. epidermidis was strongly affected by all enzyme treatments, which in addition to impairing adhesion to glass, also prevented the formation of aggregates and streamers observed abundantly in control samples. One...

  18. Glycosaminoglycan polysaccharide biosynthesis and production: today and tomorrow.

    Science.gov (United States)

    DeAngelis, Paul L

    2012-04-01

    Glycosaminoglycans [GAGs] are essential heteropolysaccharides in vertebrate tissues that are also, in certain cases, employed as virulence factors by microbes. Hyaluronan [HA], heparin, and chondroitin sulfate [CS] are GAGs currently used in various medical applications and together are multi-billion dollar products thus targets for production by animal-free manufacture. By using bacteria as the source of GAGs, the pathogen's sword may be converted into a plowshare to help avoid potential liabilities springing from the use of animal-derived GAGs including adventitious agents (e.g., prions, pathogens), antigenicity, degradation of the environment, and depletion of endangered species. HA from microbes, which have a chemical structure identical to human HA, has already been commercialized and sold at the ton-scale. Substantial progress towards microbial heparin and CS has been made, but these vertebrate polymers are more complicated structurally than the unsulfated bacterial polysaccharide precursors thus require additional processing steps. This review provides an overview of GAG structure, medical applications, microbial biosynthesis, and the state of bacterial GAG production systems. Representatives of all glycosyltransferase enzymes that polymerize the sugar chains of the three main GAGs have been identified and serve as the core technology to harness, but the proteins involved in sugar precursor formation and chain export steps of biosynthesis are also essential to the GAG production process. In addition, this review discusses future directions and potential important issues. Overall, this area is poised to make great headway to produce safer (both increased purity and more secure supply chains) non-animal GAG-based therapeutics.

  19. Algal Production of Extra- and Intra-Cellular Polysaccharides as an Adaptive Response to the Toxin Crude Extract of Microcystis Aeruginosa

    Directory of Open Access Journals (Sweden)

    Mostafa Mohamed El-Sheekh

    2012-11-01

    Full Text Available This is an investigation concerned with studying the possible adaptive response of four different unicellular algae, Anabaena PCC 7120, Oscillatoria angustissima, Scendesmus obliquus and Chlorella vulgaris, to the toxin of Microcystis aeruginosa (Kützing. Theeffects of four different concentrations, 25, 50, 100 and 200 μg mL-1 of microcystins crude extract of M. aeruginosa, on both intra and extra-cellular polysaccharide levels, in log phase,of the four tested algae were studied. The obtained results showed differential increase in the production levels for both intra and extra-cellular polysaccharides by the tested algae,compared with the control. S. obliquus and C. vulgaris showed a resistance to crude toxinhigher than Anabaena PCC 7120 and O. angustissima. The highly production of polysaccharides by green algal species under this toxic stress indicated the involvement of these polysaccharides in protecting the algal cells against toxic species and, reflect thebiological behavior of particular algal species to the environmental stresses.

  20. Algal production of extra and intra-cellular polysaccharides as an adaptive response to the toxin crude extract of Microcystis aeruginosa

    Directory of Open Access Journals (Sweden)

    El-Sheekh Mostafa

    2012-11-01

    Full Text Available Abstract This is an investigation concerned with studying the possible adaptive response of four different unicellular algae, Anabaena PCC 7120, Oscillatoria angustissima, Scendesmus obliquus and Chlorella vulgaris, to the toxin of Microcystis aeruginosa (Kützing. The effects of four different concentrations, 25, 50, 100 and 200 μg mL-1 of microcystins crude extract of M. aeruginosa, on both intra and extra-cellular polysaccharide levels, in log phase, of the four tested algae were studied. The obtained results showed differential increase in the production levels for both intra and extra-cellular polysaccharides by the tested algae, compared with the control. S. obliquus and C. vulgaris showed a resistance to crude toxin higher than Anabaena PCC 7120 and O. angustissima. The highly production of polysaccharides by green algal species under this toxic stress indicated the involvement of these polysaccharides in protecting the algal cells against toxic species and, reflect the biological behavior of particular algal species to the environmental stresses.

  1. Application of high OLR-fed aerobic granules for the treatment of low-strength wastewater: Performance, granule morphology and microbial community

    Institute of Scientific and Technical Information of China (English)

    Jingyun Ma; Xiangchun Quan; Huai Li

    2013-01-01

    Aerobic granules,pre-cultivated at the organic loading rate (OLR) of 3.0 kg COD/(m3·day),were used to treat low-strength wastewater in two sequencing batch reactors at low OLRs of 1.2 and 0.6 kg COD/(m3·day),respectively.Reactor performance,evolution of granule morphology,structure and microbial community at low OLRs under long-term operation (130 days) were investigated.Results showed that low OLRs did not cause significant damage to granule structure as a dominant granule morphology with size over 540 μm was maintained throughout the operation.Aerobic granules at sizes of about 750 μm were finally obtained at the low OLRs.The granule reactors operated at low OLRs demonstrated effective COD and ammonia removals (above 90%),smaller granule sizes and less biomass.The contents of extracellular polymeric substances in the granules were decreased while the ratios of exopolysaccharide/exoprotein were increased (above 1.0).The granules cultivated at the low OLRs showed a smoother surface and more compact structure than the seeded granules.A significant shift in microbial community was observed but the microbial diversity remained relatively stable.Confocal Laser Scanning Microscopy observation showed that the live cells were spread throughout the whole granule,while the dead cells were mainly concentrated in the outer layer of the granule,and the proteins,polysaccharides and lipids were mainly located in the central regime of the granule.In conclusion,granules cultivated at high OLRs show potential for treating low-strength organic wastewater steadily under long-term operation.

  2. Extracellular Proteins Limit the Dispersal of BiogenicNanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Moreau, John W.; Weber, Peter K.; Martin, Michael C.; Gilbert,Benjamin; Hutcheon, Ian D.; Banfield, Jillian F.

    2007-04-27

    High spatial-resolution secondaryion microprobespectrometry, synchrotron radiation Fourier-transform infraredspectroscopy and polyacrylamide gel analysis demonstrate the intimateassociation of proteins with spheroidal aggregates of biogenic zincsulfide nanocrystals, an example of extracellular biomineralization.Experiments involving synthetic ZnS nanoparticles and representativeamino acids indicate a driving role for cysteine in rapid nanoparticleaggregation. These findings suggest that microbially-derivedextracellular proteins can limit dispersal of nanoparticulatemetal-bearing phases, such as the mineral products of bioremediation,that may otherwise be transported away from their source by subsurfacefluid flow.

  3. Identification of enzymes responsible for extracellular alginate depolymerization and alginate metabolism in Vibrio algivorus.

    Science.gov (United States)

    Doi, Hidetaka; Tokura, Yuriko; Mori, Yukiko; Mori, Kenichi; Asakura, Yoko; Usuda, Yoshihiro; Fukuda, Hiroo; Chinen, Akito

    2017-02-01

    Alginate is a marine non-food-competing polysaccharide that has potential applications in biorefinery. Owing to its large size (molecular weight >300,000 Da), alginate cannot pass through the bacterial cell membrane. Therefore, bacteria that utilize alginate are presumed to have an enzyme that degrades extracellular alginate. Recently, Vibrio algivorus sp. SA2(T) was identified as a novel alginate-decomposing and alginate-utilizing species. However, little is known about the mechanism of alginate degradation and metabolism in this species. To address this issue, we screened the V. algivorus genomic DNA library for genes encoding polysaccharide-decomposing enzymes using a novel double-layer plate screening method and identified alyB as a candidate. Most identified alginate-decomposing enzymes (i.e., alginate lyases) must be concentrated and purified before extracellular alginate depolymerization. AlyB of V. algivorus heterologously expressed in Escherichia coli depolymerized extracellular alginate without requiring concentration or purification. We found seven homologues in the V. algivorus genome (alyB, alyD, oalA, oalB, oalC, dehR, and toaA) that are thought to encode enzymes responsible for alginate transport and metabolism. Introducing these genes into E. coli enabled the cells to assimilate soluble alginate depolymerized by V. algivorus AlyB as the sole carbon source. The alginate was bioconverted into L-lysine (43.3 mg/l) in E. coli strain AJIK01. These findings demonstrate a simple and novel screening method for identifying polysaccharide-degrading enzymes in bacteria and provide a simple alginate biocatalyst and fermentation system with potential applications in industrial biorefinery.

  4. Ganoderma lucidum polysaccharide exerts anti-tumor activity via MAPK pathways in HL-60 acute leukemia cells.

    Science.gov (United States)

    Yang, Guohua; Yang, Lei; Zhuang, Yun; Qian, Xifeng; Shen, Yunfeng

    2016-01-01

    In this study, we investigated the anti-tumor activity both in vitro and in vivo of a polysaccharide obtained from Ganoderma lucidum on HL-60 acute myeloid leukemia cells, and focused on its targeting effect on mitogen-activated protein kinase (MAPK) pathways. It was found by the methods such as western blot and flow cytometry (FCM), that G. lucidum polysaccharide (GLP) blocked the extracellular signal-regulated kinase/MAPK signaling pathway, simultaneously activated p38 and JNK MAPK pathways, and therefore regulated their downstream genes and proteins, including p53, c-myc, c-fos, c-jun, Bcl-2, Bax, cleaved caspase-3 and cyclin D1. As a result, cycle arrest and apoptosis of HL-60 cells were induced. Therefore, GLP exerted anti-tumor activity via MAPK pathways in HL-60 acute leukemia cells.

  5. Bacterial capsular polysaccharide prevents the onset of asthma through T-cell activation.

    Science.gov (United States)

    Johnson, Jenny L; Jones, Mark B; Cobb, Brian A

    2015-04-01

    Over the last four decades, increases in the incidence of immune-mediated diseases in the Western world have been linked to changes in microbial exposure. It is becoming increasingly clear that the normal microbiota in the gut can profoundly alter susceptibility to a wide range of diseases, such as asthma, in which immune homeostasis is disrupted, yet the mechanisms governing this microbial influence remains poorly defined. In this study, we show that gastrointestinal exposure to PSA, a capsular polysaccharide derived from the commensal bacterium Bacteroides fragilis, significantly limits susceptibility to the induction of experimental asthma. We report that direct treatment of mice with PSA generates protection from asthma, and this effect can be given to a naïve recipient by adoptive transfer of CD4(+) T cells from PSA-exposed mice. Remarkably, we found that these PSA-induced T cells are not canonical FoxP3(+) regulatory T cells, but that they potently inhibit both Th1 and Th2 models of asthma in an IL-10-dependent fashion. These findings reveal that bacterial polysaccharides link the microbiota with the peripheral immune system by activating CD4(+)Foxp3(-) T cells upon exposure in the gut, and they facilitate resistance to unnecessary inflammatory responses via the production of IL-10. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Complete structure of the cell surface polysaccharide of Streptococcus oralis C104: A 600-MHz NMR study

    Energy Technology Data Exchange (ETDEWEB)

    Abeygunawardana, C.; Bush, C.A. (Univ. of Maryland, Baltimore (United States)); Cisar, J.O. (National Inst. of Dental Research, Bethesda, MD (United States))

    1991-09-03

    Specific lectin-carbohydrate interactions between certain oral streptococci and actinomyces contribute to the microbial colonization of teeth. The receptor molecules of Streptococcus oralis, 34, ATCC 10557, and Streptococcus mitis J22 for the galactose and N-acetylgalactosamine reactive fimbrial lectins of Actinomyces viscosus and Actinomyces naeslundii are antigenically distinct polysaccharides, each formed by a different phosphodiester-linked oligosaccharide repeating unit. Receptor polysaccharide was isolated form S. oralis C104 cells and was shown to contain galactose, N-acetylgalactosamine, ribitol, and phosphate with molar ratios of 4:1:1:1. The {sup 1}H NMR spectrum of the polysaccharide shows that it contains a repeating structure. The individual sugars in the repeating unit were identified by {sup 1}H coupling constants observed in E-COSY and DQF-COSY spectra. NMR methods included complete resonance assignments ({sup 1}H and {sup 13}C) by various homonuclear and heteronuclear correlation experiments that utilize scalar couplings. Sequence and linkage assignments were obtained from the heteronuclear multiple-bond correlation (HMBC) spectrum. This analysis shows that the receptor polysaccharide of S. oralis C104 is a ribitol teichoic acid polymer composed of a linear hexasaccharide repeating unit containing two residues each of galactopyranose and galactofuranose and a residue each of GalNAc and ribitol joined end to end by phosphodiester linkages.

  7. In vitro fermentation characteristics of two mushroom species, an herb, and their polysaccharide fractions, using chicken cecal contents as inoculum.

    Science.gov (United States)

    Guo, F C; Williams, B A; Kwakkel, R P; Verstegen, M W A

    2003-10-01

    In vitro fermentabilities of two mushrooms (Lentinus edodes--LenS; Tremella fuciformis--TreS), an herb (Astragalus membranaceus--AstS), and their polysaccharide fractions (LenE, TreE, and AstE) were investigated using microflora from chicken ceca. Polysaccharides were extracted using the hot water method. The mushrooms had lower polysaccharide yields (8 to 10%) than the herb (31%). Fermentation kinetics were determined using the in vitro cumulative gas production technique. End-products, such as gas, volatile fatty acids (VFA), and ammonia, were also determined. The gas profiles of intact materials were similar for AstS and LenS. The TreS had a diphasic digestion pattern. The extracts had similar profiles to the intact materials though gas production rates were faster. Intact materials tended to produce less VFA than the extracts though LenS and AstE had the highest total VFA production overall. Intact materials contained more protein than the extracts, and therefore resulted in more branched-chain fatty acids and ammonia. Fermentation kinetics and end-point products demonstrated differences in availability of substrates between the mushrooms and herb. These medicinal mushroom and herb materials, particularly their polysaccharide extracts, show promise in altering microbial activities and composition in chicken ceca. In vivo experiments are necessary for confirmation of this hypothesis.

  8. A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns.

    Science.gov (United States)

    Laezza, Antonio; Iadonisi, Alfonso; Pirozzi, Anna V A; Diana, Paola; De Rosa, Mario; Schiraldi, Chiara; Parrilli, Michelangelo; Bedini, Emiliano

    2016-12-12

    Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. To this aim, a modular approach based on the different combination of only five reactions was employed, with an almost unprecedented polysaccharide branching by O-glycosylation as the key step. The library was differentiated for sulfation patterns and/or positions of the fucose branches, as confirmed by detailed 2D NMR spectroscopic analysis. These semi-synthetic polysaccharides will allow a wider and more accurate structure-activity relationship study with respect to those reported in literature to date.

  9. The dietary polysaccharide maltodextrin promotes Salmonella survival and mucosal colonization in mice.

    Directory of Open Access Journals (Sweden)

    Kourtney P Nickerson

    Full Text Available In the latter half of the 20th century, societal and technological changes led to a shift in the composition of the American diet to include a greater proportion of processed, pre-packaged foods high in fat and carbohydrates, and low in dietary fiber (a "Western diet". Over the same time period, there have been parallel increases in Salmonella gastroenteritis cases and a broad range of chronic inflammatory diseases associated with intestinal dysbiosis. Several polysaccharide food additives are linked to bacterially-driven intestinal inflammation and may contribute to the pathogenic effects of a Western diet. Therefore, we examined the effect of a ubiquitous polysaccharide food additive, maltodextrin (MDX, on clearance of the enteric pathogen Salmonella using both in vitro and in vivo infection models. When examined in vitro, murine bone marrow-derived macrophages exposed to MDX had altered vesicular trafficking, suppressed NAPDH oxidase expression, and reduced recruitment of NADPH oxidase to Salmonella-containing vesicles, which resulted in persistence of Salmonella in enlarged Rab7+ late endosomal vesicles. In vivo, mice consuming MDX-supplemented water had a breakdown of the anti-microbial mucous layer separating gut bacteria from the intestinal epithelium surface. Additionally, oral infection of these mice with Salmonella resulted in increased cecal bacterial loads and enrichment of lamina propria cells harboring large Rab7+ vesicles. These findings indicate that consumption of processed foods containing the polysaccharide MDX contributes to suppression of intestinal anti-microbial defense mechanisms and may be an environmental priming factor for the development of chronic inflammatory disease.

  10. Extracellular DNA chelates cations and induces antibiotic resistance in Pseudomonas aeruginosa biofilms.

    Directory of Open Access Journals (Sweden)

    Heidi Mulcahy

    2008-11-01

    Full Text Available Biofilms are surface-adhered bacterial communities encased in an extracellular matrix composed of DNA, bacterial polysaccharides and proteins, which are up to 1000-fold more antibiotic resistant than planktonic cultures. To date, extracellular DNA has been shown to function as a structural support to maintain Pseudomonas aeruginosa biofilm architecture. Here we show that DNA is a multifaceted component of P. aeruginosa biofilms. At physiologically relevant concentrations, extracellular DNA has antimicrobial activity, causing cell lysis by chelating cations that stabilize lipopolysaccharide (LPS and the outer membrane (OM. DNA-mediated killing occurred within minutes, as a result of perturbation of both the outer and inner membrane (IM and the release of cytoplasmic contents, including genomic DNA. Sub-inhibitory concentrations of DNA created a cation-limited environment that resulted in induction of the PhoPQ- and PmrAB-regulated cationic antimicrobial peptide resistance operon PA3552-PA3559 in P. aeruginosa. Furthermore, DNA-induced expression of this operon resulted in up to 2560-fold increased resistance to cationic antimicrobial peptides and 640-fold increased resistance to aminoglycosides, but had no effect on beta-lactam and fluoroquinolone resistance. Thus, the presence of extracellular DNA in the biofilm matrix contributes to cation gradients, genomic DNA release and inducible antibiotic resistance. DNA-rich environments, including biofilms and other infection sites like the CF lung, are likely the in vivo environments where extracellular pathogens such as P. aeruginosa encounter cation limitation.

  11. Anodic and cathodic microbial communities in single chamber microbial fuel cells.

    Science.gov (United States)

    Daghio, Matteo; Gandolfi, Isabella; Bestetti, Giuseppina; Franzetti, Andrea; Guerrini, Edoardo; Cristiani, Pierangela

    2015-01-25

    Microbial fuel cells (MFCs) are a rapidly growing technology for energy production from wastewater and biomasses. In a MFC, a microbial biofilm oxidizes organic matter and transfers electrons from reduced compounds to an anode as the electron acceptor by extracellular electron transfer (EET). The aim of this work was to characterize the microbial communities operating in a Single Chamber Microbial Fuel Cell (SCMFC) fed with acetate and inoculated with a biogas digestate in order to gain more insight into anodic and cathodic EET. Taxonomic characterization of the communities was carried out by Illumina sequencing of a fragment of the 16S rRNA gene. Microorganisms belonging to Geovibrio genus and purple non-sulfur (PNS) bacteria were found to be dominant in the anodic biofilm. The alkaliphilic genus Nitrincola and anaerobic microorganisms belonging to Porphyromonadaceae family were the most abundant bacteria in the cathodic biofilm.

  12. Microbial Metalloproteomics

    Directory of Open Access Journals (Sweden)

    Peter-Leon Hagedoorn

    2015-12-01

    Full Text Available Metalloproteomics is a rapidly developing field of science that involves the comprehensive analysis of all metal-containing or metal-binding proteins in a biological sample. The purpose of this review is to offer a comprehensive overview of the research involving approaches that can be categorized as inductively coupled plasma (ICP-MS based methods, X-ray absorption/fluorescence, radionuclide based methods and bioinformatics. Important discoveries in microbial proteomics will be reviewed, as well as the outlook to new emerging approaches and research areas.

  13. Microbial exopolysaccharides as determinants of geomorphological, hydrological and optical properties of soil crusts from the Precambrian till today

    Science.gov (United States)

    Garcia-Pichel, F.

    2012-04-01

    The presence of microbial extracellular polysaccharides (EPS) in the soil solution and/or in association with particular microbial types can impart novel properties to biological soil crust (BSC), and hence to soil surfaces. For the most part these properties are of a geobiological relevance that exceeds what one could surmise from its relatively low specific mass content. I will review some examples that range from the mundane to the unexpected. EPS associated with filamentous cyanobacteria can effectively and in the long term stabilize the soil surface against erosive forces, even after the microbes are long gone. Electrostatic interactions between EPS and blowing dust may help retain dust particles, enriching the soil with new nutrient sources. In a telltale sign of BSC presence, EPS is the agent that allows sandy soils to fold and curl-up, to form pee-tee's and elephant-skin surfaces, and to crack into polygons like clays would. EPS in large quantities in flat crusts can retain fluids (both liquid and gaseous) resulting in the alteration of hydrological flow and in the formation of internal vesicular horizons, gas bubbles, pock-marked surfaces and other characteristic structures. Yet, in some settings, EPS plays an architectural role in creating a "spongy" texture that increases hydraulic conductivity. This architectural role can indirectly result in significant increases of a crust's albedo. While the diversity of consequences of EPS presence is far from understood, evidence for its sustained role through Earth's history can be found in the form of sedimentary bio-signatures as far back as the Proterozoic.

  14. α-Amylase-assisted extraction of polysaccharides from Panax ginseng.

    Science.gov (United States)

    Sun, Lin; Wu, Di; Ning, Xin; Yang, Guang; Lin, Ziheng; Tian, Meihong; Zhou, Yifa

    2015-04-01

    In this paper, α-amylase-assisted extraction was used to isolate the polysaccharide that remained in hot water-extracted ginseng. The yield of the polysaccharide was 9.0%, almost equal to that of the hot water-extracted polysaccharide. Using anion exchange and gel permeation chromatography, the polysaccharide was fractionated into a neutral polysaccharide fraction and six pectic fractions. The neutral fraction accounted for 76% of the polysaccharide and contained both amylopectin and amylose. The pectic polysaccharide fractions were identified to be arabinogalactan, type-I rhamnogalacturonan and homogalacturonan-type pectin by high-performance liquid chromatography, Fourier transform-infrared and nuclear magnetic resonance analysis. Structural and lymphocyte proliferation activity results showed that these polysaccharides were different from those extracted by hot water, indicating that ginseng contains complex polysaccharides with diverse structures, which results in its diverse pharmacological activities. The α-amylase-assisted extraction is a novel method for preparing ginseng polysaccharides and could be applied toward the further study and exploration of ginseng. These findings provide technical and theoretical support for ginseng pharmacology. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Microbial Ecosystems, Protection of

    NARCIS (Netherlands)

    Bodelier, P.L.E.; Nelson, K.E.

    2014-01-01

    Synonyms Conservation of microbial diversity and ecosystem functions provided by microbes; Preservation of microbial diversity and ecosystem functions provided by microbes Definition The use, management, and conservation of ecosystems in order to preserve microbial diversity and functioning.

  16. Microbial Ecosystems, Protection of

    NARCIS (Netherlands)

    Bodelier, P.L.E.; Nelson, K.E.

    2014-01-01

    Synonyms Conservation of microbial diversity and ecosystem functions provided by microbes; Preservation of microbial diversity and ecosystem functions provided by microbes Definition The use, management, and conservation of ecosystems in order to preserve microbial diversity and functioning. Introdu

  17. Magnetite Compensates for the Lack of a Pilin-Associated c-Type Cytochrome in Extracellular Electron Exchange

    DEFF Research Database (Denmark)

    Liu, Fanghua; Rotaru, Amelia-Elena; Shrestha, Pravin;

    2015-01-01

    Nano-scale magnetite can facilitate microbial extracellular electron transfer that plays an important role in biogeochemical cycles, bioremediation, and several bioenergy strategies, but the mechanisms for the stimulation of extracellular electron transfer are poorly understood. Further investiga......Nano-scale magnetite can facilitate microbial extracellular electron transfer that plays an important role in biogeochemical cycles, bioremediation, and several bioenergy strategies, but the mechanisms for the stimulation of extracellular electron transfer are poorly understood. Further......S-deficient strain unable to participate in interspecies electron transfer or Fe(III) oxide reduction. In the presence of magnetite wild-type cells repressed expression of the OmcS gene, suggesting that cells might need to produce less OmcS when magnetite was available. The finding that magnetite can compensate...

  18. PssP2 is a polysaccharide co-polymerase involved in exopolysaccharide chain-length determination in Rhizobium leguminosarum.

    Directory of Open Access Journals (Sweden)

    Małgorzata Marczak

    Full Text Available Production of extracellular polysaccharides is a complex process engaging proteins localized in different subcellular compartments, yet communicating with each other or even directly interacting in multicomponent complexes. Proteins involved in polymerization and transport of exopolysaccharide (EPS in Rhizobium leguminosarum are encoded within the chromosomal Pss-I cluster. However, genes implicated in polysaccharide synthesis are common in rhizobia, with several homologues of pss genes identified in other regions of the R. leguminosarum genome. One such region is chromosomally located Pss-II encoding proteins homologous to known components of the Wzx/Wzy-dependent polysaccharide synthesis and transport systems. The pssP2 gene encodes a protein similar to polysaccharide co-polymerases involved in determination of the length of polysaccharide chains in capsule and O-antigen biosynthesis. In this work, a mutant with a disrupted pssP2 gene was constructed and its capabilities to produce EPS and enter into a symbiotic relationship with clover were studied. The pssP2 mutant, while not altered in lipopolysaccharide (LPS, displayed changes in molecular mass distribution profile of EPS. Lack of the full-length PssP2 protein resulted in a reduction of high molecular weight EPS, yet polymerized to a longer length than in the RtTA1 wild type. The mutant strain was also more efficient in symbiotic performance. The functional interrelation between PssP2 and proteins encoded within the Pss-I region was further supported by data from bacterial two-hybrid assays providing evidence for PssP2 interactions with PssT polymerase, as well as glycosyltransferase PssC. A possible role for PssP2 in a complex involved in EPS chain-length determination is discussed.

  19. Extracellular vesicles for drug delivery

    NARCIS (Netherlands)

    Vader, Pieter; Mol, Emma A; Pasterkamp, Gerard; Schiffelers, Raymond M

    2016-01-01

    Extracellular vesicles (EVs) are cell-derived membrane vesicles, and represent an endogenous mechanism for intercellular communication. Since the discovery that EVs are capable of functionally transferring biological information, the potential use of EVs as drug delivery vehicles has gained consider

  20. Antihypertensive activity of polysaccharide from Crassostrea gigas.

    Science.gov (United States)

    Wang, Ting; Ding, Jie; Li, Haibo; Xiang, Jingjing; Wen, Ping; Zhang, Qin; Yin, Linliang; Jiang, Wei; Shen, Caie

    2016-02-01

    Water-soluble polysaccharide was extracted from Crassostrea gigas by hydrolysis with flavourzyme and filtered, ultrafiltered and precipitated using absolute ethanol. Sugar composition analysis performed on the C. gigas polysaccharide (CGP) by high performance liquid chromatography indicated that it was comprised primarily of glucose, and its molecular weight was determined using a TSK-GEL G5000PW column to be ∼3.413×10(6) Da. Next, the antihypertensive activity of CGP was evaluated in rats. Hypertension model Wistar rats were divided into three groups and intragastrically treated with physiological saline (negative control group), CGP (treatment group), and captopril (positive control group). CGP treatment led to significant decrease in both systolic and diastolic pressures in the hypertension model Wistar rats. Furthermore, the antihypertensive effect of CGP was comparable with that of captopril. Thus, CGP has antihypertensive effects and can potentially be used as a therapeutic agent for hypertension. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Structural characterization of polysaccharides from bamboo

    Science.gov (United States)

    Kamil, Ruzaimah Nik Mohamad; Yusuf, Nur'aini Raman; Yunus, Normawati M.; Yusup, Suzana

    2014-10-01

    The alkaline and water soluble polysaccharides were isolate by sequential extractions with distilled water, 60% ethanol containing 1%, 5% and 8% NaOH. The samples were prepared at 60 °C for 3 h from local bamboo. The functional group of the sample were examined using FTIR analysis. The most precipitate obtained is from using 60% ethanol containing 8% NaOH with yield of 2.6%. The former 3 residues isolated by sequential extractions with distilled water, 60% ethanol containing 1% and 5% NaOH are barely visible after filtering with cellulose filter paper. The FTIR result showed that the water-soluble polysaccharides consisted mainly of OH group, CH group, CO indicates the carbohydrate and sugar chain. The sample weight loss was slightly decreased with increasing of temperature.

  2. Lytic polysaccharide monooxygenases: a crystallographer’s view on a new class of biomass-degrading enzymes

    Science.gov (United States)

    Frandsen, Kristian E. H.; Lo Leggio, Leila

    2016-01-01

    Lytic polysaccharide monooxygenases (LPMOs) are a new class of microbial copper enzymes involved in the degradation of recalcitrant polysaccharides. They have only been discovered and characterized in the last 5–10 years and have stimulated strong interest both in biotechnology and in bioinorganic chemistry. In biotechnology, the hope is that these enzymes will finally help to make enzymatic biomass conversion, especially of lignocellulosic plant waste, economically attractive. Here, the role of LPMOs is likely to be in attacking bonds that are not accessible to other enzymes. LPMOs have attracted enormous interest since their discovery. The emphasis in this review is on the past and present contribution of crystallographic studies as a guide to functional understanding, with a final look towards the future. PMID:27840684

  3. Pneumococcal capsular polysaccharide structure predicts serotype prevalence.

    Directory of Open Access Journals (Sweden)

    Daniel M Weinberger

    2009-06-01

    Full Text Available There are 91 known capsular serotypes of Streptococcus pneumoniae. The nasopharyngeal carriage prevalence of particular serotypes is relatively stable worldwide, but the host and bacterial factors that maintain these patterns are poorly understood. Given the possibility of serotype replacement following vaccination against seven clinically important serotypes, it is increasingly important to understand these factors. We hypothesized that the biochemical structure of the capsular polysaccharides could influence the degree of encapsulation of different serotypes, their susceptibility to killing by neutrophils, and ultimately their success during nasopharyngeal carriage. We sought to measure biological differences among capsular serotypes that may account for epidemiological patterns. Using an in vitro assay with both isogenic capsule-switch variants and clinical carriage isolates, we found an association between increased carriage prevalence and resistance to non-opsonic neutrophil-mediated killing, and serotypes that were resistant to neutrophil-mediated killing tended to be more heavily encapsulated, as determined by FITC-dextran exclusion. Next, we identified a link between polysaccharide structure and carriage prevalence. Significantly, non-vaccine serotypes that have become common in vaccinated populations tend to be those with fewer carbons per repeat unit and low energy expended per repeat unit, suggesting a novel biological principle to explain patterns of serotype replacement. More prevalent serotypes are more heavily encapsulated and more resistant to neutrophil-mediated killing, and these phenotypes are associated with the structure of the capsular polysaccharide, suggesting a direct relationship between polysaccharide biochemistry and the success of a serotype during nasopharyngeal carriage and potentially providing a method for predicting serotype replacement.

  4. INTERACTION OF IONIC LIQUIDS WITH POLYSACCHARIDES

    OpenAIRE

    2008-01-01

    The use of ionic liquids (ILs) in the field of cellulose chemistry opens up a broad variety of new opportunities. Besides the regeneration of the biopolymer to fibers, films, and beads, this new class of cellulose solvents is particularly useful for the homogeneous chemical modification of the polysaccharide. In this review, the potential of ILs as a reaction medium for the homogeneous cellulose functionalization is discussed. It is shown that numerous conversions proceed very efficiently and...

  5. Ratiometric imaging of extracellular pH in bacterial biofilms with C-SNARF-4.

    Science.gov (United States)

    Schlafer, Sebastian; Garcia, Javier E; Greve, Matilde; Raarup, Merete K; Nyvad, Bente; Dige, Irene

    2015-02-01

    pH in the extracellular matrix of bacterial biofilms is of central importance for microbial metabolism. Biofilms possess a complex three-dimensional architecture characterized by chemically different microenvironments in close proximity. For decades, pH measurements in biofilms have been limited to monitoring bulk pH with electrodes. Although pH microelectrodes with a better spatial resolution have been developed, they do not permit the monitoring of horizontal pH gradients in biofilms in real time. Quantitative fluorescence microscopy can overcome these problems, but none of the hitherto employed methods differentiated accurately between extracellular and intracellular microbial pH and visualized extracellular pH in all areas of the biofilms. Here, we developed a method to reliably monitor extracellular biofilm pH microscopically with the ratiometric pH-sensitive dye C-SNARF-4, choosing dental biofilms as an example. Fluorescent emissions of C-SNARF-4 can be used to calculate extracellular pH irrespective of the dye concentration. We showed that at pH values of biofilm and visualized the entire bacterial biomass in in vivo-grown dental biofilms with unknown species composition. We then employed digital image analysis to remove the bacterial biomass from the microscopic images and adequately calculate extracellular pH values. As a proof of concept, we monitored the extracellular pH drop in in vivo-grown dental biofilms fermenting glucose. The combination of pH ratiometry with C-SNARF-4 and digital image analysis allows the accurate monitoring of extracellular pH in bacterial biofilms in three dimensions in real time and represents a significant improvement to previously employed methods of biofilm pH measurement.

  6. Marine Origin Polysaccharides in Drug Delivery Systems

    Directory of Open Access Journals (Sweden)

    Matias J. Cardoso

    2016-02-01

    Full Text Available Oceans are a vast source of natural substances. In them, we find various compounds with wide biotechnological and biomedical applicabilities. The exploitation of the sea as a renewable source of biocompounds can have a positive impact on the development of new systems and devices for biomedical applications. Marine polysaccharides are among the most abundant materials in the seas, which contributes to a decrease of the extraction costs, besides their solubility behavior in aqueous solvents and extraction media, and their interaction with other biocompounds. Polysaccharides such as alginate, carrageenan and fucoidan can be extracted from algae, whereas chitosan and hyaluronan can be obtained from animal sources. Most marine polysaccharides have important biological properties such as biocompatibility, biodegradability, and anti-inflammatory activity, as well as adhesive and antimicrobial actions. Moreover, they can be modified in order to allow processing them into various shapes and sizes and may exhibit response dependence to external stimuli, such as pH and temperature. Due to these properties, these biomaterials have been studied as raw material for the construction of carrier devices for drugs, including particles, capsules and hydrogels. The devices are designed to achieve a controlled release of therapeutic agents in an attempt to fight against serious diseases, and to be used in advanced therapies, such as gene delivery or regenerative medicine.

  7. Plant Cell Wall Matrix Polysaccharide Biosynthesis

    Institute of Scientific and Technical Information of China (English)

    Ajay Pal S. Sandhu; Gursharn S. Randhawa; Kanwarpal S. Dhugga

    2009-01-01

    The wall of an expanding plant cell consists primarily of cellulose microfibrils embedded in a matrix of hemi-cellulosic and pectic polysaccharides along with small amounts of structural and enzymatic proteins. Matrix polysacchar-ides are synthesized in the Golgi and exported to the cell wall by exocytosis, where they intercalate among cellulose microfibrUs, which are made at the plasma membrane and directly deposited into the cell wall. Involvement of Golgi glucan synthesis in auxin-induced cell expansion has long been recognized; however, only recently have the genes corresponding to glucan synthases been identified. Biochemical purification was unsuccessful because of the labile nature and very low abundance of these enzymes. Mutational genetics also proved fruitless. Expression of candidate genes identified through gene expression profiling or comparative genomics in heterologous systems followed by functional characterization has been relatively successful. Several genes from the cellulose synthase-like (Cs/) family have been found to be involved in the synthesis of various hemicellulosic glycans. The usefulness of this approach, however, is limited to those enzymes that probably do not form complexes consisting of unrelated proteins. Nonconventional approaches will continue to incre-mentally unravel the mechanisms of Golgi polysaccharide biosynthesis.

  8. Rheological studies of polysaccharides for skin scaffolds.

    Science.gov (United States)

    Almeida, Nalinda; Mueller, Anja; Hirschi, Stanley; Rakesh, Leela

    2014-05-01

    Polysaccharide hydrogels are good candidates for skin scaffolds because of their inherent biocompatibility and water transport properties. In the current study, hydrogels were made from a mixture of four polysaccharides: xanthan gum, konjac gum, iota-carrageenan, and kappa-carrageenan. Gel formation, strength, and structure of these polysaccharides were studied using rheological and thermal techniques. All gel samples studied were strong gels at all times because of the gradual water loss. However, after 12 h of storage, elastic (G') and loss (G'') moduli of hydrogel mixture containing all the ingredients is of one to two orders of magnitude greater than that of mixtures not containing either xanthan gum or iota-carrageenan, which confirmed the varied levels of gel strength. This is mainly due to the rate of water loss in each of these mixtures, resulting in gels of varying structures and dynamic moduli over a period of time. Iota-carrageenan and xanthan gum differ in their effect on gel strength and stability in combination with konjac gum and kappa-carrageenan.

  9. Synthetic Polymer Nanoparticle-Polysaccharide Interactions: A Systematic Study

    OpenAIRE

    Zeng, Zhiyang; Patel, Jiten; Lee, Shih-Hui; McCallum, Monica; Tyagi, Anuradha; Yan, Mingdi; Shea, Kenneth J.

    2012-01-01

    The interaction between synthetic polymer nanoparticles (NPs) and biomacromolecules (e.g. proteins, lipids and polysaccharides) can profoundly influence the NPs fate and function. Polysaccharides (e.g. heparin/heparin sulfate) are a key component of cell surfaces and the extracelluar matrix and play critical roles in many biological processes. We report a systematic investigation of the interaction between synthetic polymer nanoparticles and polysaccharides by ITC, SPR and an anticoagulant as...

  10. Structural Analysis of Polysaccharide from Cladonia furcatar and Its Activities

    Institute of Scientific and Technical Information of China (English)

    CHEN; Qian

    2001-01-01

    Cladonia furcatar is a member of the lichens genus Cladonia. Japanese scientific researchers [1] made a great achievement in selecting lichens as anticancer agent and polysaccharides are generally considered to be their main bioactive constituents. Recently, Iacomini et al[2] have investigated the members of the genus Cladonia, a mannose-containing polysaccharide was isolated from Cladoniafurcatar. We now report the new chemical structure of polysaccharide from Cladoniafurcatar and its activities.  ……

  11. Structural Analysis of Polysaccharide from Cladonia furcatar and Its Activities

    Institute of Scientific and Technical Information of China (English)

    CHEN Qian; LIN Xin; CAI YuJun; LI ZhiXiao

    2001-01-01

    @@ Cladonia furcatar is a member of the lichens genus Cladonia. Japanese scientific researchers [1] made a great achievement in selecting lichens as anticancer agent and polysaccharides are generally considered to be their main bioactive constituents. Recently, Iacomini et al[2] have investigated the members of the genus Cladonia, a mannose-containing polysaccharide was isolated from Cladoniafurcatar. We now report the new chemical structure of polysaccharide from Cladoniafurcatar and its activities.

  12. Measurement of microbial growth in the low nutrient conditions of a simulated subsurface environment

    Science.gov (United States)

    Hand, V. L.; Boult, S.; Vaughan, D. J.; Beadle, I. R.; Humphreys, P.; Wogelius, R. A.

    2003-04-01

    The growth of bacteria in natural porous media may alter porosity and permeability, and therefore hydraulic conductivity. Changes are due both to pore clogging caused by the production of bacterial extracellular polysaccharides (EPS) and the formation of secondary mineral precipitates. Pore clogging has implications not only for fluid flow, but also for contaminant transport. Most biofilm research has been conducted under nutrient rich conditions, quite different from the actual subsurface environment. There is therefore a general need for studies under environmentally relevant conditions. The main objectives were to determine growth under environmental conditions and to produce reproducible homogeneously coated columns of porous media for further experiments on metal transport. Six short columns (length 25mm; diameter 20mm) instrumented with pressure, pH and dissolved oxygen sensors were used. Growth and reproducibility of the biofilm are related to the flow rate, the concentration of the nutrients and the grain size of the porous medium substrate. Two types of porous media were used; a single mineral quartz media with a constant grain size and a natural mixed mineral assemblage of non-uniform grain size. Nutrient used was a landfill derived carbon source present in synthetic trench leachate (STL) diluted with synthetic groundwater (SGW) by a factor of 100. The STL was pumped through each column at a constant, environmentally relevant flow rate of 0.109 ml/min. Experiments were performed under both aerobic and anaerobic conditions contained in a temperature-controlled room at 10^oC. Measurement of the pressure increase within the column apparatus was made, as an increase in pressure is relative to the resistance of the media to flow and indicates biofilm formation and pore clogging within the column. Monitoring of dissolved oxygen shows the metabolic conditions of the bacteria and also the biological oxygen demand. Column effluent was analysed for changes in

  13. Preservation and Significance of Extracellular DNA in Ferruginous Sediments from Lake Towuti, Indonesia

    Science.gov (United States)

    Vuillemin, Aurèle; Horn, Fabian; Alawi, Mashal; Henny, Cynthia; Wagner, Dirk; Crowe, Sean A.; Kallmeyer, Jens

    2017-01-01

    Extracellular DNA is ubiquitous in soil and sediment and constitutes a dominant fraction of environmental DNA in aquatic systems. In theory, extracellular DNA is composed of genomic elements persisting at different degrees of preservation produced by processes occurring on land, in the water column and sediment. Extracellular DNA can be taken up as a nutrient source, excreted or degraded by microorganisms, or adsorbed onto mineral matrices, thus potentially preserving information from past environments. To test whether extracellular DNA records lacustrine conditions, we sequentially extracted extracellular and intracellular DNA from anoxic sediments of ferruginous Lake Towuti, Indonesia. We applied 16S rRNA gene Illumina sequencing on both fractions to discriminate exogenous from endogenous sources of extracellular DNA in the sediment. Environmental sequences exclusively found as extracellular DNA in the sediment originated from multiple sources. For instance, Actinobacteria, Verrucomicrobia, and Acidobacteria derived from soils in the catchment. Limited primary productivity in the water column resulted in few sequences of Cyanobacteria in the oxic photic zone, whereas stratification of the water body mainly led to secondary production by aerobic and anaerobic heterotrophs. Chloroflexi and Planctomycetes, the main degraders of sinking organic matter and planktonic sequences at the water-sediment interface, were preferentially preserved during the initial phase of burial. To trace endogenous sources of extracellular DNA, we used relative abundances of taxa in the intracellular DNA to define which microbial populations grow, decline or persist at low density with sediment depth. Cell lysis became an important additional source of extracellular DNA, gradually covering previous genetic assemblages as other microbial genera became more abundant with depth. The use of extracellular DNA as nutrient by active microorganisms led to selective removal of sequences with lowest

  14. Preservation and Significance of Extracellular DNA in Ferruginous Sediments from Lake Towuti, Indonesia

    Directory of Open Access Journals (Sweden)

    Aurèle Vuillemin

    2017-07-01

    Full Text Available Extracellular DNA is ubiquitous in soil and sediment and constitutes a dominant fraction of environmental DNA in aquatic systems. In theory, extracellular DNA is composed of genomic elements persisting at different degrees of preservation produced by processes occurring on land, in the water column and sediment. Extracellular DNA can be taken up as a nutrient source, excreted or degraded by microorganisms, or adsorbed onto mineral matrices, thus potentially preserving information from past environments. To test whether extracellular DNA records lacustrine conditions, we sequentially extracted extracellular and intracellular DNA from anoxic sediments of ferruginous Lake Towuti, Indonesia. We applied 16S rRNA gene Illumina sequencing on both fractions to discriminate exogenous from endogenous sources of extracellular DNA in the sediment. Environmental sequences exclusively found as extracellular DNA in the sediment originated from multiple sources. For instance, Actinobacteria, Verrucomicrobia, and Acidobacteria derived from soils in the catchment. Limited primary productivity in the water column resulted in few sequences of Cyanobacteria in the oxic photic zone, whereas stratification of the water body mainly led to secondary production by aerobic and anaerobic heterotrophs. Chloroflexi and Planctomycetes, the main degraders of sinking organic matter and planktonic sequences at the water-sediment interface, were preferentially preserved during the initial phase of burial. To trace endogenous sources of extracellular DNA, we used relative abundances of taxa in the intracellular DNA to define which microbial populations grow, decline or persist at low density with sediment depth. Cell lysis became an important additional source of extracellular DNA, gradually covering previous genetic assemblages as other microbial genera became more abundant with depth. The use of extracellular DNA as nutrient by active microorganisms led to selective removal of

  15. Antiviral polysaccharides isolated from Hong Kong brown seaweed Hydroclathrus clathratus

    Institute of Scientific and Technical Information of China (English)

    WANG; Hui; OOI; Engchoon; Vincent; ANG; Put; O; Jr.

    2007-01-01

    Two relatively pure polysaccharides H3-a1 and H3-b1 had been isolated from the brown seaweed Hydroclathrus clathratus. They were characterized by HPLC, ultraviolet scanning, gas chromatography, infrared spectroscopy and elemental analysis, and shown to be two different sulfated polysaccharides with different monosaccharide content, but both with high relative molecular mass. They contained some proteins and uronic acid respectively. The sulfate content and bioactivity of these polysaccharides varied during purification. The fractions derived from the hot water extract also exhibited low anticoagulant effect. This is the first time that the antiherpetic and anticoagulant activities were evaluated for the polysaccharides from the Hong Kong brown seaweed Hydroclathrus clathratus.

  16. Effect of free ammonium and free nitrous acid on the activity, aggregate morphology and extracellular polymeric substance distribution of ammonium oxidizing bacteria in partial nitrification.

    Science.gov (United States)

    Yao, Qian; Peng, Dangcong; Wang, Bo; Chen, Yuanyuan; Li, Jiaqi; Zhao, Qiaodi; Wang, Binbin

    2017-09-01

    Successful partial nitrification not only guarantees the inhibition of nitrite oxidation, but also does not excessively retard the ammonia oxidation rate. Therefore, the performance of ammonium oxidizing bacteria (AOB) during partial nitrification is fundamental to this process. In this study, two lab-scale partial nitrification bioreactors containing different inhibition conditions-one with free ammonium (FA) inhibition, the other with free nitrous acid (FNA) inhibition-were used to compare the differences between activity, quantity, aggregation morphology and extracellular polymeric substance (EPS) distribution of AOB. The results showed that although stable, long-term, partial nitrification was achieved in both reactors, there were differences in AOB activity, microbial spatial distribution and EPS characteristic. In the FA bioreactor, FA concentration was conducted at more than 40 mg/L, which had a strong impact on the metabolism of AOB. The activity and quantity decreased by 50%. Higher EPS (42.44 ± 2.31 mg g(-1) mixed liquor volatile suspended solids [MLVSS]) and protein were introduced into the EPS matrix. However, in the FNA bioreactor, the FNA concentration was about 0.23 mg/L. It did not reach a level to affect AOB metabolism. The AOB activity and quantity were maintained at high levels and the total EPS content was 28.29 ± 2.04 mg g(-1) MLVSS. Additionally, the microscopic results showed that in the FA bioreactor, AOB cells aggregated in microcolonies, while they appeared to be self-flocculating with no specific conformation in the other reactor. β-polysaccharides located inside sludge flocs in the FA bioreactor but only accumulated around the outer layer of activated sludge flocs in the FNA condition. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  17. Heparan sulphate, its derivatives and analogues share structural characteristics that can be exploited, particularly in inhibiting microbial attachment

    Directory of Open Access Journals (Sweden)

    T.R. Rudd

    2012-05-01

    Full Text Available Heparan sulphate (HS and the related polysaccharide, heparin, exhibit conformational and charge arrangement properties, which provide a degree of redundancy allowing several seemingly distinct sequences to exhibit the same activity. This can also be mimicked by other sulphated polysaccharides, both in overall effect and in the details of interactions and structural consequences of interactions with proteins. Together, these provide a source of active compounds suitable for further development as potential drugs. These polysaccharides also possess considerable size, which bestows upon them an additional useful property: the capability of disrupting processes comprising many individual interactions, such as those characterising the attachment of microbial pathogens to host cells. The range of involvement of HS in microbial attachment is reviewed and examples, which include viral, bacterial and parasitic infections and which, in many cases, are now being investigated as potential targets for intervention, are identified.

  18. Study of extracellular polymeric substances in the biofilms of a suspended biofilter for nitric oxide removal.

    Science.gov (United States)

    Li, Han; Huang, Shaobin; Zhou, Shaofeng; Chen, Pengfei; Zhang, Yongqing

    2016-11-01

    The extraction and quantitative analysis of extracellular polymeric substances (EPS) have been frequently reported in studies of activated sludge. However, little is currently known about the EPS in the biofilms of biofilter systems. This study investigates the EPS in biofilms of Chelatococcus daeguensis TAD1 established in a suspended biofilter for nitric oxide (NO) removal under thermophilic conditions. Polysaccharide was the main EPS component under all experimental operation conditions of the aerobic biofilter, although the EPS contents and components varied under different operating conditions. As the concentration of the inlet NO varied from 200 to 2000 mg/m(3), the EPS and protein contents generally increased. At the highest inlet concentration (2000 mg/m(3)), the EPS and protein contents reached 0.118 and 0.055 mg/g, respectively (representing increases of 7.3 and 35 %, respectively, over the inlet concentration of 200 mg/m(3)). In contrast, the polysaccharide content was quite stable against inlet NO concentration. Decreasing the empty bed residence time increased the EPS and polysaccharide contents, but exerted little effect on the protein content. Varying the pH of the circulating fluid from 4 to 8 changed the EPS and its components in complex ways. We also found a strong correlation between the total EPS content and the NO removal efficiency. Therefore, it is possible to take EPS into consideration for biofilter control.

  19. Review of Micro/Nanotechnologies for Microbial Biosensors

    Directory of Open Access Journals (Sweden)

    Ji Won eLim

    2015-05-01

    Full Text Available A microbial biosensor is an analytical device with a biologically integrated transducer that generates a measurable signal indicating the analyte concentration. This method is ideally suited for the analysis of extracellular chemicals and the environment, and for metabolic sensory-regulation. Although microbial biosensors show promise for application in various detection fields, some limitations still remain such as poor selectivity, low sensitivity, and impractical portability. To overcome such limitations, microbial biosensors have been integrated with many recently developed micro/nanotechnologies and applied to a wide range of detection purposes. This review article discusses micro/nanotechnologies that have been integrated with microbial biosensors and summarizes recent advances and the applications achieved through such novel integration. Future perspectives on the combination of micro/nanotechnologies and microbial biosensors will be discussed, and the necessary developments and improvements will be strategically deliberated.

  20. Extracellular vesicles: Exosomes, microvesicles, and friends

    NARCIS (Netherlands)

    Raposo, G.; Stoorvogel, W.|info:eu-repo/dai/nl/074352385

    2013-01-01

    Cells release into the extracellular environment diverse types of membrane vesicles of endosomal and plasma membrane origin called exosomes and microvesicles, respectively. These extracellular vesicles (EVs) represent an important mode of intercellular communication by serving as vehicles for

  1. Streptococcus mutans-derived extracellular matrix in cariogenic oral biofilms.

    Science.gov (United States)

    Klein, Marlise I; Hwang, Geelsu; Santos, Paulo H S; Campanella, Osvaldo H; Koo, Hyun

    2015-01-01

    Biofilms are highly structured microbial communities that are enmeshed in a self-produced extracellular matrix. Within the complex oral microbiome, Streptococcus mutans is a major producer of extracellular polymeric substances including exopolysaccharides (EPS), eDNA, and lipoteichoic acid (LTA). EPS produced by S. mutans-derived exoenzymes promote local accumulation of microbes on the teeth, while forming a spatially heterogeneous and diffusion-limiting matrix that protects embedded bacteria. The EPS-rich matrix provides mechanical stability/cohesiveness and facilitates the creation of highly acidic microenvironments, which are critical for the pathogenesis of dental caries. In parallel, S. mutans also releases eDNA and LTA, which can contribute with matrix development. eDNA enhances EPS (glucan) synthesis locally, increasing the adhesion of S. mutans to saliva-coated apatitic surfaces and the assembly of highly cohesive biofilms. eDNA and other extracellular substances, acting in concert with EPS, may impact the functional properties of the matrix and the virulence of cariogenic biofilms. Enhanced understanding about the assembly principles of the matrix may lead to efficacious approaches to control biofilm-related diseases.

  2. Streptococcus mutans-derived extracellular matrix in cariogenic oral biofilms

    Directory of Open Access Journals (Sweden)

    Marlise eKlein

    2015-02-01

    Full Text Available Biofilms are highly structured microbial communities that are enmeshed in a self-produced extracellular matrix. Within the complex oral microbiome, Streptococcus mutans is a major producer of extracellular polymeric substances including exopolysaccharides (EPS, eDNA and lipoteichoic acid (LTA. EPS produced by S. mutans-derived exoenzymes promote local accumulation of microbes on the teeth, while forming a spatially heterogeneous and diffusion-limiting matrix that protects embedded bacteria. The EPS-rich matrix provides mechanical stability/cohesiveness and facilitates the creation of highly acidic microenvironments, which are critical for the pathogenesis of dental caries. In parallel, S. mutans also releases eDNA and LTA, which can contribute with matrix development. eDNA enhances EPS (glucan synthesis locally, increasing the adhesion of S. mutans to saliva-coated apatitic surfaces and the assembly of highly cohesive biofilms. eDNA and other extracellular substances, acting in concert with EPS, may impact the functional properties of the matrix and the virulence of cariogenic biofilms. Enhanced understanding about the assembly principles of the matrix may lead to efficacious approaches to control biofilm-related diseases.

  3. Neutrophil Extracellular Traps and Microcrystals

    Science.gov (United States)

    2017-01-01

    Neutrophil extracellular traps represent a fascinating mechanism by which PMNs entrap extracellular microbes. The primary purpose of this innate immune mechanism is thought to localize the infection at an early stage. Interestingly, the ability of different microcrystals to induce NET formation has been recently described. Microcrystals are insoluble crystals with a size of 1–100 micrometers that have different composition and shape. Microcrystals have it in common that they irritate phagocytes including PMNs and typically trigger an inflammatory response. This review is the first to summarize observations with regard to PMN activation and NET release induced by microcrystals. Gout-causing monosodium urate crystals, pseudogout-causing calcium pyrophosphate dehydrate crystals, cholesterol crystals associated with atherosclerosis, silicosis-causing silica crystals, and adjuvant alum crystals are discussed. PMID:28373994

  4. Neutrophil Extracellular Traps and Microcrystals.

    Science.gov (United States)

    Rada, Balázs

    2017-01-01

    Neutrophil extracellular traps represent a fascinating mechanism by which PMNs entrap extracellular microbes. The primary purpose of this innate immune mechanism is thought to localize the infection at an early stage. Interestingly, the ability of different microcrystals to induce NET formation has been recently described. Microcrystals are insoluble crystals with a size of 1-100 micrometers that have different composition and shape. Microcrystals have it in common that they irritate phagocytes including PMNs and typically trigger an inflammatory response. This review is the first to summarize observations with regard to PMN activation and NET release induced by microcrystals. Gout-causing monosodium urate crystals, pseudogout-causing calcium pyrophosphate dehydrate crystals, cholesterol crystals associated with atherosclerosis, silicosis-causing silica crystals, and adjuvant alum crystals are discussed.

  5. Neutrophil Extracellular Traps and Microcrystals

    Directory of Open Access Journals (Sweden)

    Balázs Rada

    2017-01-01

    Full Text Available Neutrophil extracellular traps represent a fascinating mechanism by which PMNs entrap extracellular microbes. The primary purpose of this innate immune mechanism is thought to localize the infection at an early stage. Interestingly, the ability of different microcrystals to induce NET formation has been recently described. Microcrystals are insoluble crystals with a size of 1–100 micrometers that have different composition and shape. Microcrystals have it in common that they irritate phagocytes including PMNs and typically trigger an inflammatory response. This review is the first to summarize observations with regard to PMN activation and NET release induced by microcrystals. Gout-causing monosodium urate crystals, pseudogout-causing calcium pyrophosphate dehydrate crystals, cholesterol crystals associated with atherosclerosis, silicosis-causing silica crystals, and adjuvant alum crystals are discussed.

  6. Extracellular secretion of recombinant proteins

    Science.gov (United States)

    Linger, Jeffrey G.; Darzins, Aldis

    2014-07-22

    Nucleic acids encoding secretion signals, expression vectors containing the nucleic acids, and host cells containing the expression vectors are disclosed. Also disclosed are polypeptides that contain the secretion signals and methods of producing polypeptides, including methods of directing the extracellular secretion of the polypeptides. Exemplary embodiments include cellulase proteins fused to secretion signals, methods to produce and isolate these polypeptides, and methods to degrade lignocellulosic biomass.

  7. [Insertional mutation in the AZOBR_p60120 gene is accompanied by defects in the synthesis of lipopolysaccharide and calcofluor-binding polysaccharides in the bacterium Azospirillum brasilense Sp245].

    Science.gov (United States)

    Katsy, E I; Prilipov, A G

    2015-03-01

    In the bacterium Azospirillum brasilense Sp245, extracellular calcofluor-binding polysaccharides (Cal+ phenotype) and two types of lipopolysaccharides, LPSI and LPSII, were previously identified. These lipopolysaccharides share the same repeating O-polysaccharide unit but have different antigenic structures and different charges of their O-polysaccharides and/or core oligosaccharides. Several dozens of predicted genes involved in the biosynthesis of polysaccharides have been localized in the AZOBR_p6 plasmid of strain Sp245 (GenBank accession no. HE577333). In the present work, it was demonstrated that an artificial transposon Omegon-Km had inserted into the central region of the AZOBR_p60120 gene in the A. brasilense Sp245 LPSI- Cal- KM252 mutant. In A. brasilense strain Sp245, this plasmid gene encodes a putative glycosyltransferase containing conserved domains characteristic of the enzymes participating in the synthesis of O-polysaccharides and capsular polysaccharides (accession no. YP004987664). In mutant KM252, a respective predicted protein is expected to be completely inactivated. As a result of the analysis of the EcoRI fragment of the AZOBR_p6 plasmid, encompassing the AZOBR_p60120 gene and a number of other loci, novel data on the structure of AZOBR_p6 were obtained: an approximately 5-kb gap (GenBank accession no. KM189439) was closed in the nucleotide sequence of this plasmid.

  8. Aggregation and removal of copper oxide (CuO) nanoparticles in wastewater environment and their effects on the microbial activities of wastewater biofilms.

    Science.gov (United States)

    Miao, Lingzhan; Wang, Chao; Hou, Jun; Wang, Peifang; Ao, Yanhui; Li, Yi; Geng, Nan; Yao, Yu; Lv, Bowen; Yang, Yangyang; You, Guoxiang; Xu, Yi

    2016-09-01

    The transport behaviors of copper oxide (CuO) NPs in wastewater matrix and their possible impacts on microbial activities of stable wastewater biofilms cultivated in a lab scale rotating biological contactor (RBC) were investigated. Significant aggregation of CuO NPs was observed in the wastewater samples, depending on their mass concentrations. Extracellular polymeric substance (EPS)-adsorbed copper accounted for a large proportion of the total copper accumulated in biofilms. The microelectrode profiles showed that a single pulse exposure to 50mg/L CuO resulted in a deeper penetration depth of oxygen in biofilms compared to the CuO NP free biofilms. The maximum oxygen consumption rate shifted to the deeper parts of biofilms, indicating that the respiration activities of bacteria in the top region of the biofilms was significantly inhibited by CuO NPs. Biofilms secreted more EPS in response to the nano-CuO stress, with higher production of proteins compared to polysaccharides. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Evidence for marine origin and microbial-viral habitability of sub-zero hypersaline aqueous inclusions within permafrost near Barrow, Alaska.

    Science.gov (United States)

    Colangelo-Lillis, J; Eicken, H; Carpenter, S D; Deming, J W

    2016-05-01

    Cryopegs are sub-surface hypersaline brines at sub-zero temperatures within permafrost; their global extent and distribution are unknown. The permafrost barrier to surface and groundwater advection maintains these brines as semi-isolated systems over geological time. A cryopeg 7 m below ground near Barrow, Alaska, was sampled for geochemical and microbiological analysis. Sub-surface brines (in situtemperature of -6 °C, salinity of 115 ppt), and an associated sediment-infused ice wedge (melt salinity of 0.04 ppt) were sampled using sterile technique. Major ionic concentrations in the brine corresponded more closely to other (Siberian) cryopegs than to Standard seawater or the ice wedge. Ionic ratios and stable isotope analysis of water conformed to a marine or brackish origin with subsequent Rayleigh fractionation. The brine contained ∼1000× more bacteria than surrounding ice, relatively high viral numbers suggestive of infection and reproduction, and an unusually high ratio of particulate to dissolved extracellular polysaccharide substances. A viral metagenome indicated a high frequency of temperate viruses and limited viral diversity compared to surface environments, with closest similarity to low water activity environments. Interpretations of the results underscore the isolation of these underexplored microbial ecosystems from past and present oceans.

  10. Immunomodulatory dietary polysaccharides: a systematic review of the literature

    Directory of Open Access Journals (Sweden)

    Nelson Erika D

    2010-11-01

    Full Text Available Abstract Background A large body of literature suggests that certain polysaccharides affect immune system function. Much of this literature, however, consists of in vitro studies or studies in which polysaccharides were injected. Their immunologic effects following oral administration is less clear. The purpose of this systematic review was to consolidate and evaluate the available data regarding the specific immunologic effects of dietary polysaccharides. Methods Studies were identified by conducting PubMed and Google Scholar electronic searches and through reviews of polysaccharide article bibliographies. Only articles published in English were included in this review. Two researchers reviewed data on study design, control, sample size, results, and nature of outcome measures. Subsequent searches were conducted to gather information about polysaccharide safety, structure and composition, and disposition. Results We found 62 publications reporting statistically significant effects of orally ingested glucans, pectins, heteroglycans, glucomannans, fucoidans, galactomannans, arabinogalactans and mixed polysaccharide products in rodents. Fifteen controlled human studies reported that oral glucans, arabinogalactans, heteroglycans, and fucoidans exerted significant effects. Although some studies investigated anti-inflammatory effects, most studies investigated the ability of oral polysaccharides to stimulate the immune system. These studies, as well as safety and toxicity studies, suggest that these polysaccharide products appear to be largely well-tolerated. Conclusions Taken as a whole, the oral polysaccharide literature is highly heterogenous and is not sufficient to support broad product structure/function generalizations. Numerous dietary polysaccharides, particularly glucans, appear to elicit diverse immunomodulatory effects in numerous animal tissues, including the blood, GI tract and spleen. Glucan extracts from the Trametes versicolor

  11. Induction of cellulases and hemicellulases by tamarind (Tamarindus indica) kernel polysaccharide

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh, B.S.; Kundu, A.B.

    1980-01-01

    Tamarind kernel polysaccharide (TKP) which is available in India in abundance could be used as an excellent substrate for production of cellulases, hemicellulases, ..beta..-glucosidase and ..beta..-xylosidase. A growth medium of a known cellulytic strain, Aspergillus terreus containing TKP has been optimized to get enhanced yields of cellulase and hemicellulase for practical application in jute manufacturing units around Calcutta. Of the large number of sugars and treated and untreated cellulosics tested, TKP was found to produce the highest amounts of cellulases, hemicellulases, ..beta..-glucosidase, ..beta..-xylosidase and extra-cellular protein. The effects of the use of different inorganic nitrogenous substances, nutrients and surfactants in the optimized medium have been tested. Both initial pH of the medium and final pH of the culture filtrate have been found to have a marked effect on enzyme production, especially ..beta..-glucosidase production.

  12. Studies on the immuno-modulating and anti-tumor activities of Ganoderma lucidum (Reishi) polysaccharides.

    Science.gov (United States)

    Chen, Hung-Sen; Tsai, Yow-Fu; Lin, Steven; Lin, Chia-Ching; Khoo, Kay-Hooi; Lin, Chun-Hung; Wong, Chi-Huey

    2004-11-01

    We describe here the isolation of Reishi polysaccharides for the study of their effect on cytokine expression in mouse splenocytes. A fraction (F3) has been shown to activate the expression of IL-1, IL-6, IL-12, IFN-gamma, TNF-alpha, GM-CSF, G-CSF, and M-CSF, and from this three subfractions have been prepared where F3G1 activates IL-1, IL-12, TNF-alpha, and G-CSF, F3G2 activates all the cytokines as F3 does, and F3G3 activates only IL-1 and TNF-alpha. Together with previous studies, the mode of action on macrophages has been proposed where F3 binds to TLR4 receptor and activates extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 to induce IL-1 expression.

  13. Effect of the Polysaccharide Extract from the Edible Mushroom Pleurotus ostreatus against Infectious Bursal Disease Virus

    Directory of Open Access Journals (Sweden)

    Tatiana Rugea

    2009-08-01

    Full Text Available The polysaccharide-containing extracellular fractions (EFs of the edible mushroom Pleurotus ostreatus have immunomodulating effects. Being aware of these therapeutic effects of mushroom extracts, we have investigated the synergistic relations between these extracts and BIAVAC and BIAROMVAC vaccines. These vaccines target the stimulation of the immune system in commercial poultry, which are extremely vulnerable in the first days of their lives. By administrating EF with polysaccharides from P. ostreatus to unvaccinated broilers we have noticed slow stimulation of maternal antibodies against infectious bursal disease (IBD starting from four weeks post hatching. For the broilers vaccinated with BIAVAC and BIAROMVAC vaccines a low to almost complete lack of IBD maternal antibodies has been recorded. By adding 5% and 15% EF in the water intake, as compared to the reaction of the immune system in the previous experiment, the level of IBD antibodies was increased. This has led us to believe that by using this combination of BIAVAC and BIAROMVAC vaccine and EF from P. ostreatus we can obtain good results in stimulating the production of IBD antibodies in the period of the chicken first days of life, which are critical to broilers’ survival. This can be rationalized by the newly proposed reactivity biological activity (ReBiAc principles by examining the parabolic relationship between EF administration and recorded biological activity.

  14. Effect of the Polysaccharide Extract from the Edible Mushroom Pleurotus ostreatus against Infectious Bursal Disease Virus

    Science.gov (United States)

    Selegean, Mircea; Putz, Mihai V.; Rugea, Tatiana

    2009-01-01

    The polysaccharide-containing extracellular fractions (EFs) of the edible mushroom Pleurotus ostreatus have immunomodulating effects. Being aware of these therapeutic effects of mushroom extracts, we have investigated the synergistic relations between these extracts and BIAVAC and BIAROMVAC vaccines. These vaccines target the stimulation of the immune system in commercial poultry, which are extremely vulnerable in the first days of their lives. By administrating EF with polysaccharides from P. ostreatus to unvaccinated broilers we have noticed slow stimulation of maternal antibodies against infectious bursal disease (IBD) starting from four weeks post hatching. For the broilers vaccinated with BIAVAC and BIAROMVAC vaccines a low to almost complete lack of IBD maternal antibodies has been recorded. By adding 5% and 15% EF in the water intake, as compared to the reaction of the immune system in the previous experiment, the level of IBD antibodies was increased. This has led us to believe that by using this combination of BIAVAC and BIAROMVAC vaccine and EF from P. ostreatus we can obtain good results in stimulating the production of IBD antibodies in the period of the chicken first days of life, which are critical to broilers’ survival. This can be rationalized by the newly proposed reactivity biological activity (ReBiAc) principles by examining the parabolic relationship between EF administration and recorded biological activity. PMID:20111675

  15. Stratification of extracellular polymeric substances (EPS) for aggregated anammox microorganisms.

    Science.gov (United States)

    Jia, Fangxu; Yang, Qing; Liu, Xiuhong; Li, Xiyao; Li, Baikun; Zhang, Liang; Peng, Yongzhen

    2017-02-27

    Sludge aggregation and biofilm formation are the most effective approaches to solve the washout of anammox microorganisms. In this study, the structure and composition of EPS (extracellular polymeric substances) were investigated to elucidate the factors for the anammox aggregation property. Anammox sludge taken from 18 lab-scale and pilot-scale reactors treating different types of wastewater was analyzed using EEM-PARAFAC (excitation-emission matrix and parallel factor analysis), FTIR (fourier transform infrared spectroscopy) and real-time PCR combined with multivariate statistical analysis. The results showed that slime and TB-EPS (tightly bound EPS) were closely related with water quality and sludge morphology, and could be used as the indicators for anammox microbial survival ability and microbial aggregate morphology. Furthermore, slime secreted from anammox bacterial cells may be exhibited higher viscosity to the sludge surface and easily formed the gel network to aggregate. Large amounts of hydrophobic groups of protein in TB-EPS promoted the microbial aggregation. The mechanisms of anammox aggregation explored in this study enhanced the understanding of anammox stability in wastewater treatment processes.

  16. Deposition kinetics of extracellular polymeric substances (EPS) on silica in monovalent and divalent salts.

    Science.gov (United States)

    Zhu, Pingting; Long, Guoyu; Ni, Jinren; Tong, Meiping

    2009-08-01

    The deposition kinetics of extracellular polymeric substances (EPS) on silica surfaces were examined in both monovalent and divalent solutions under a variety of environmentally relevant ionic strength and pH conditions by employing a quartz crystal microbalance with dissipation (DCM-D). Soluble EPS (SEPS) and bound EPS (BEPS) were extracted from four bacterial strains with different characteristics. Maximum favorable deposition rates (k(fa)) were observed for all EPS at low ionic strengths in both NaCl and CaCl2 solutions. With the increase of ionic strength, k(fa) decreased due to the simultaneous occurrence of EPS aggregation in solutions. Deposition efficiency (alpha; the ratio of deposition rates obtained under unfavorable versus corresponding favorable conditions) for all EPS increased with increasing ionic strength in both NaCl and CaCl2 solutions, which agreed with the trends of zeta potentials and was consistent with the classic Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. Comparison of alpha for SEPS and BEPS extracted from the same strain showed that the trends of alpha did not totally agree with trends of zeta potentials, indicating the deposition kinetics of EPS on silica surfaces were not only controlled by DLVO interactions, but also non-DLVO forces. Close comparison of alpha for EPS extracted from different sources showed alpha increased with increasing proteins to polysaccharides ratio. Subsequent experiments for EPS extracted from the same strain but with different proteins to polysaccharides ratios and from activated sludge also showed that alpha were largest for EPS with greatest proteins to polysaccharides ratio. Additional experiments for pure protein and solutions with different pure proteins to pure saccharides ratios further corroborated that larger proteins to polysaccharides ratio resulted in greater EPS deposition.

  17. Methods to identify the unexplored diversity of microbial exopolysaccharides.

    Science.gov (United States)

    Rühmann, Broder; Schmid, Jochen; Sieber, Volker

    2015-01-01

    Microbial exopolysaccharides (EPS) are a structurally very diverse class of molecules. A number of them have found their application in rather diverging fields that extend from medicine, food, and cosmetics on the one side to construction, drilling, and chemical industry on the other side. The analysis of microbial strains for their competence in polysaccharide production has therefore been a major issue in the past, especially in the search for new polysaccharide variants among natural strain isolates. Concerning the fact that nearly all microbes carry the genetic equipment for the production of polysaccharides under specific conditions, the naturally provided EPS portfolio seems to be still massively underexplored. Therefore, there is a need for high throughput screening techniques capable of identifying novel variants of bacterial EPS with properties superior to the already described ones, or even totally new ones. A great variety of different techniques has been used in screening approaches for identifying microorganisms that are producing EPS in substantial amounts. Mucoid growth is often the method of choice for visual identification of EPS producing strains. Depending on the thickening characteristics of the polysaccharide, observation of viscosity in culture broth can also be an option to evaluate EPS production. Precipitation with different alcohols represents a common detection, isolation, and purification method for many EPS. A more quantitative approach is found in the total carbohydrate content analysis, normally determined, e.g., by phenol-sulfuric-acid-method. In addition, recently a new and reliable method for the detailed analysis of the monomeric composition and the presence of rare sugars and sugar substitutions has become available, which could give a first hint of the polymer structure of unknown EPS. This minireview will compare available methods and novel techniques and discuss their benefits and disadvantages.

  18. Methods to identify the unexplored diversity of microbial exopolysaccharides

    Directory of Open Access Journals (Sweden)

    Broder eRühmann

    2015-06-01

    Full Text Available Microbial exopolysaccharides (EPS are a structurally very diverse class of molecules. A number of them have found their application in rather diverging fields that extend from medicine, food and cosmetics on the one side to construction, drilling and chemical industry on the other side. The analysis of microbial strains for their competence in polysaccharide production has therefore been a major issue in the past, especially in the search for new polysaccharide variants among natural strain isolates. Concerning the fact that nearly all microbes carry the genetic equipment for the production of polysaccharides under specific conditions, the naturally provided EPS portfolio seems to be still massively underexplored. Therefore, there is a need for high throughput screening techniques capable of identifying novel variants of bacterial exopolysaccharides with properties superior to the already described ones, or even totally new ones. A great variety of different techniques has been used in screening approaches for identifying microorganisms that are producing EPS in substantial amounts. Mucoid growth is often the method of choice for visual identification of EPS producing strains. Depending on the thickening characteristics of the polysaccharide, observation of viscosity in culture broth can also be an option to evaluate EPS production. Precipitation with different alcohols represents a common detection, isolation and purification method for many EPS. A more quantitative approach is found in the total carbohydrate content analysis, normally determined e.g. by phenol-sulfuric-acid-method. In addition, recently a new and reliable method for the detailed analysis of the monomeric composition and the presence of rare sugars and sugar substitutions has become available, which could give a first hint of the polymer structure of unknown EPS. This minireview will compare available methods and novel techniques and discuss their benefits and disadvantages.

  19. Polysaccharides of Aloe vera induce MMP-3 and TIMP-2 gene expression during the skin wound repair of rat.

    Science.gov (United States)

    Tabandeh, Mohammad Reza; Oryan, Ahmad; Mohammadalipour, Adel

    2014-04-01

    Polysaccharides are the main macromolecules of Aloe vera gel but no data about their effect on extracellular matrix (ECM) elements are available. Here, mannose rich Aloe vera polysaccharides (AVP) with molecular weight between 50 and 250 kDa were isolated and characterized. Open cutaneous wounds on the back of 45 rats (control and treated) were daily treated with 25mg (n=15) and 50 mg (n=15) AVP for 30 days. The levels of MMP-3 and TIMP-2 gene expression were analyzed using real time PCR. The levels of n-acetyl glucosamine (NAGA), n-acetyl galactosamine (NAGLA) and collagen contents were also measured using standard biochemical methods. Faster wound closure was observed at day 15 post wounding in AVP treated animals in comparison with untreated group. At day 10 post wounding, AVP inhibited MMP-3 gene expression, while afterwards MMP-3 gene expression was upregulated. AVP enhanced TIMP-2 gene expression, collagen, NAGLA and NAGA synthesis in relation to untreated wounds. Our results suggest that AVP has positive effects on the regulation of ECM factor synthesis, which open up new perspectives for the wound repair activity of Aloe vera polysaccharide at molecular level.

  20. Methods of saccharification of polysaccharides in plants

    Science.gov (United States)

    Howard, John; Fake, Gina

    2014-04-29

    Saccharification of polysaccharides of plants is provided, where release of fermentable sugars from cellulose is obtained by adding plant tissue composition. Production of glucose is obtained without the need to add additional .beta.-glucosidase. Adding plant tissue composition to a process using a cellulose degrading composition to degrade cellulose results in an increase in the production of fermentable sugars compared to a process in which plant tissue composition is not added. Using plant tissue composition in a process using a cellulose degrading enzyme composition to degrade cellulose results in decrease in the amount of cellulose degrading enzyme composition or exogenously applied cellulase required to produce fermentable sugars.

  1. INTERACTION OF IONIC LIQUIDS WITH POLYSACCHARIDES

    Directory of Open Access Journals (Sweden)

    Tim Liebert

    2008-05-01

    Full Text Available The use of ionic liquids (ILs in the field of cellulose chemistry opens up a broad variety of new opportunities. Besides the regeneration of the biopolymer to fibers, films, and beads, this new class of cellulose solvents is particularly useful for the homogeneous chemical modification of the polysaccharide. In this review, the potential of ILs as a reaction medium for the homogeneous cellulose functionalization is discussed. It is shown that numerous conversions proceed very efficiently and the ILs may be recycled. But it is also demonstrated that some side reactions have to be considered.

  2. Tribological properties of neutral polysaccharide solutions under simulated oral conditions

    NARCIS (Netherlands)

    Zinoviadou, K.G.; Janssen, A.M.; Jongh, H.H.J.de

    2008-01-01

    Predictability of the perception of foods thickened by polysaccharides is only poor. Therefore, the effect of saliva on the lubrication properties of 2 types of neutral polysaccharides, cross-linked starch and locust bean gum, was studied. Despite the similar bulk rheological behavior of the 2

  3. A novel capsular polysaccharide from Rhizobium rubi strain DSM 30149.

    Science.gov (United States)

    De Castro, Cristina; Fregolino, Eleonora; Gargiulo, Valentina; Lanzetta, Rosa; Parrilli, Michelangelo

    2008-07-07

    Rhizobium rubi, strain DSM 30149, is a Gram negative phytopathogenic bacterium which produces a linear polysaccharide with the following repeating unit: This new structure was determined by spectroscopical and chemical methods. It presents similar lipophilic features reported for another strain of R. rubi. These contrast with features already known for capsular polysaccharide species from symbiontic members of the Rhizobiaceae family, namely highly anionic polymers.

  4. Separation of Pneumcoccal Capsular Polysaccharide BY Gel Extraction

    Institute of Scientific and Technical Information of China (English)

    Xu Xiaoping; Huang Xinghua; Li Zhongqin; Chen Jiebo

    2004-01-01

    Separation of pneumcoccal capsular polysaccharide by gel was investigated in this paper. The gels used here were poly(acrylamide-co-sodium methacrylate)gels and prepared by free radical polymerization in aqueous solution. The experimental results indicated that gel extraction is a potential method to separate pneumcoccal capsular polysaccharide from its dilute solution.

  5. Structural Characterization and Enzymatic Modification of Soybean Polysaccharides

    DEFF Research Database (Denmark)

    Pierce, Brian; Wichmann, Jesper

    are currently limited by the material’s insol-ubility. A central hypothesis of this work was that by obtaining a more complete understanding of the structure of this material, chemical and enzymatic ap-proaches could be developed to modify the polysaccharides, creating soluble polysaccharide fractions...

  6. Enzymatic synthesis of oligo- and polysaccharide fatty acid esters

    NARCIS (Netherlands)

    Broek, van den L.A.M.; Boeriu, C.G.

    2013-01-01

    Amphiphilic oligo- and polysaccharides (e.g. polysaccharide alkyl or alkyl-aryl esters) form a new class of polymers with exceptional properties. They function as polymeric surfactants, whilst maintaining most of the properties of the starting polymeric material such as emulsifying, gelling, and

  7. In vitro antioxidant activity of polysaccharide from Gardenia jasminoides ellis

    Science.gov (United States)

    Fan, Y.; Ge, Z.; Luo, A.

    2011-01-01

    A water-soluble polysaccharide, GP, was isolated from Gardenia jasminoides Ellis through hot water extraction followed by ethanol precipitation. The in vitro free radicals scavenging tests exhibited that GP has significant scavenging abilities especially for ABTS, DPPH, and hydroxyl radicals, which suggests that the polysaccharide GP is a novel antioxidant. ?? 2011 Academic Journals.

  8. Polysaccharides isolated from Acai fruit induce innate immune responses.

    Directory of Open Access Journals (Sweden)

    Jeff Holderness

    Full Text Available The Açaí (Acai fruit is a popular nutritional supplement that purportedly enhances immune system function. These anecdotal claims are supported by limited studies describing immune responses to the Acai polyphenol fraction. Previously, we characterized γδ T cell responses to both polyphenol and polysaccharide fractions from several plant-derived nutritional supplements. Similar polyphenol and polysaccharide fractions are found in Acai fruit. Thus, we hypothesized that one or both of these fractions could activate γδ T cells. Contrary to previous reports, we did not identify agonist activity in the polyphenol fraction; however, the Acai polysaccharide fraction induced robust γδ T cell stimulatory activity in human, mouse, and bovine PBMC cultures. To characterize the immune response to Acai polysaccharides, we fractionated the crude polysaccharide preparation and tested these fractions for activity in human PBMC cultures. The largest Acai polysaccharides were the most active in vitro as indicated by activation of myeloid and γδ T cells. When delivered in vivo, Acai polysaccharide induced myeloid cell recruitment and IL-12 production. These results define innate immune responses induced by the polysaccharide component of Acai and have implications for the treatment of asthma and infectious disease.

  9. Tribological Properties of Neutral Polysaccharide Solutions under Simulated Oral Conditions

    NARCIS (Netherlands)

    Zinoviadou, K.; Janssen, A.M.; Jongh, de H.H.J.

    2008-01-01

    Predictability of the perception of foods thickened by polysaccharides is only poor. Therefore, the effect of saliva on the lubrication properties of 2 types of neutral polysaccharides, cross-linked starch and locust bean gum, was studied. Despite the similar bulk rheological behavior of the 2 polys

  10. Characterisation of cell wall polysaccharides in bilberries and black currants

    NARCIS (Netherlands)

    Hilz, H.

    2007-01-01

    During berry juice production, polysaccharides are released from the cell walls and cause thickening and high viscosity when the berries are mashed. Consequences are a low juice yield and a poor colour. This can be prevented by the use of enzymes that degrade these polysaccharides. To use these enzy

  11. Life cycle assessment of polysaccharide materials: a review

    NARCIS (Netherlands)

    Shen, L.; Patel, M.K.

    2008-01-01

    Apart from conventional uses of polysaccharide materials, such as food, clothing, paper packaging and construction, new polysaccharide products and materials have been developed. This paper reviews life cycle assessment (LCA) studies in order to gain insight of the environmental profiles of polysacc

  12. Structural modification of polysaccharides: A biochemical-genetic approach

    Science.gov (United States)

    Kern, Roger G.; Petersen, Gene R.

    1991-01-01

    Polysaccharides have a wide range of industrial and biomedical applications. An industry trend is underway towards the increased use of bacteria to produce polysaccharides. Long term goals of this work are the adaptation and enhancement of saccharide properties for electronic and optic applications. In this report we illustrate the application of enzyme-bearing bacteriophage on strains of the enteric bacterium Klebsiella pneumoniae, which produces a polysaccharide with the relatively rare rheological property of drag-reduction. This has resulted in the production of new polysaccharides with enhanced rheological properties. Our laboratory is developing techniques for processing and structurally modifying bacterial polysaccharides and oligosaccharides which comprise their basic polymeric repeat units. Our research has focused on bacteriophage which produce specific polysaccharide degrading enzymes. This has lead to the development of enzymes generated by bacteriophage as tools for polysaccharide modification and purification. These enzymes were used to efficiently convert the native material to uniform-sized high molecular weight polymers, or alternatively into high-purity oligosaccharides. Enzyme-bearing bacteriophage also serve as genetic selection tools for bacteria that produce new families of polysaccharides with modified structures.

  13. Isolation and physical characterization of an exocellular polysaccharide

    NARCIS (Netherlands)

    Tuinier, R.; Zoon, P.; Olieman, C.; Cohen Stuart, M.A.; Fleer, G.J.; Kruif, de C.G.

    1999-01-01

    The physical properties of a polysaccharide produced by the lactic acid bacterium Lactococcus lactis subsp. cremoris strain NIZO B40 were investigated. Separation of the polysaccharide from most low molar mass compounds in the culture broth was performed by filtration processes. Residual proteins an

  14. Structural characterization and bioactivities of sulfated polysaccharide from Monostroma oxyspermum.

    Science.gov (United States)

    Seedevi, Palaniappan; Moovendhan, Meivelu; Sudharsan, Sadhasivam; Vasanthkumar, Shanmugam; Srinivasan, Alagiri; Vairamani, Shanmugam; Shanmugam, Annaian

    2015-01-01

    Sulfated polysaccharide was isolated from Monostroma oxyspermum through hot water extraction, anion-exchange and gel permeation column chromatography. The sulfated polysaccharide contained 92% of carbohydrate, 0% of protein, 7.8% of uronic acid, 22% of ash and 33% of moisture respectively. The elemental composition was analyzed using CHNS/O analyzer. The molecular weight of sulfated polysaccharide determined through PAGE was found to be as 55 kDa. Monosaccharides analysis revealed that sulfated polysaccharide was composed of rhamnose, fructose, galactose, xylose, and glucose. The structural features of sulfated polysaccharide were analyzed by NMR spectroscopy. Further the sulfated polysaccharide showed total antioxidant and DPPH free radical scavenging activity were as 66.29% at 250 μg/ml and 66.83% at 160 μg/ml respectively. The sulfated polysaccharide also showed ABTS scavenging ability and reducing power were as 83.88% at 125 μg/ml and 15.81% at 400 μg/ml respectively. The anticoagulant activity was determined for human plasma with respect to Activated Partial Thromboplastin Time (APTT) and Prothrombin Time (PT) was 20.09 IU and 1.79 IU at 25 μg/ml respectively. These results indicated that the sulfated polysaccharide from M. oxyspermum had potent antioxidant and anticoagulant activities.

  15. Insight into the functionality of microbioal exo-polysaccharides by NMR and molecular modelling

    Directory of Open Access Journals (Sweden)

    Flemming Hofmann Larsen

    2015-12-01

    Full Text Available Microbial polysaccharides represent an important class of microbial polymers with diverse functions such as biofilm formation, thickening and gelling properties as well as health-promoting properties.The broad range of exopolysaccharide functionalities has sparked a renewed interest in this class of molecules. Chemical, enzymatic as well as genetic modifications by metabolic engineering can be used to create large numbers of analogous exopolysaccharide variants with respect to exopolysaccharide functionality. While this top-down approach is effective in finding new candidates for desired functionality, there seems to be a lack of the corresponding bottom-up approach. The molecular mechanisms of the desired functionalities can be established from NMR and molecular models and it is proposed that these models can be fed back into the biotechnology by using a quantitative structure-property approach. In this way it will be possible to tailor specific functionality within a given design space.This perspective will include two well-known commercial microbial exopolysaccharide examples namely gellan and diutan and show how even a limited use of multiphase NMR and molecular modelling can increase the insight into their different properties, which are based on only minor structural differences.

  16. Structural Features and Healthy Properties of Polysaccharides Occurring in Mushrooms

    Directory of Open Access Journals (Sweden)

    Eva Guillamón

    2012-12-01

    Full Text Available Polysaccharides from mushrooms have attracted a great deal of attention due to the many healthy benefits they have demonstrated, such as immunomodulation, anticancer activity, prevention and treatment of cardiovascular diseases, antiviral and antimicrobial effects, among others. Isolation and purification of polysaccharides commonly involve several steps, and different techniques are actually available in order to increase extraction yield and purity. Studies have demonstrated that the molecular structure and arrangement significantly influence the biological activity; therefore, there is a wide range of analytical techniques for the elucidation of chemical structures. Different polysaccharides have been isolated from mushrooms, most of them consisting of β-linked glucans, such as lentinan from Lentinus edodes, pleuran from Pleurotus species, schizophyllan from Schizophyllum commune, calocyban from Calocybe indica, or ganoderan and ganopoly from Ganoderma lucidum. This article reviews the main methods of polysaccharide isolation and structural characterization, as well as some of the most important polysaccharides isolated from mushrooms and the healthy benefits they provide.

  17. Characterization of polysaccharides from Ganoderma spp. using saccharide mapping.

    Science.gov (United States)

    Wu, Ding-Tao; Xie, Jing; Hu, De-Jun; Zhao, Jing; Li, Shao-Ping

    2013-09-12

    Polysaccharides from Ganoderma spp. and their adulterants were firstly investigated and compared using saccharide mapping, enzymatic (endo-1,3-β-D-glucanase and pectinase) digestion followed by polysaccharide analysis using carbohydrate gel electrophoresis analysis. The results showed that both 1,3-β-D-glucosidic and 1,4-α-D-galactosiduronic linkages were existed in Lingzhi (Ganoderma lucidum and Ganoderma sinense), and the similarity of polysaccharides from G. lucidum and G. sinense was high, which may contribute to rational use of Lingzhi. Different species of Ganoderma and their adulterants can be differentiated based on the saccharide mapping, which is helpful to well understand the structural characters of polysaccharides from different species of Ganoderma and to improve the quality control of polysaccharides in Lingzhi.

  18. The Antiviral Activities and Mechanisms of Marine Polysaccharides: An Overview

    Science.gov (United States)

    Wang, Wei; Wang, Shi-Xin; Guan, Hua-Shi

    2012-01-01

    Recently, the studies on the antiviral activities of marine natural products, especially marine polysaccharides, are attracting more and more attention all over the world. Marine-derived polysaccharides and their lower molecular weight oligosaccharide derivatives have been shown to possess a variety of antiviral activities. This paper will review the recent progress in research on the antiviral activities and the mechanisms of these polysaccharides obtained from marine organisms. In particular, it will provide an update on the antiviral actions of the sulfated polysaccharides derived from marine algae including carrageenans, alginates, and fucans, relating to their structure features and the structure–activity relationships. In addition, the recent findings on the different mechanisms of antiviral actions of marine polysaccharides and their potential for therapeutic application will also be summarized in detail. PMID:23235364

  19. The Antiviral Activities and Mechanisms of Marine Polysaccharides: An Overview

    Directory of Open Access Journals (Sweden)

    Hua-Shi Guan

    2012-12-01

    Full Text Available Recently, the studies on the antiviral activities of marine natural products, especially marine polysaccharides, are attracting more and more attention all over the world. Marine-derived polysaccharides and their lower molecular weight oligosaccharide derivatives have been shown to possess a variety of antiviral activities. This paper will review the recent progress in research on the antiviral activities and the mechanisms of these polysaccharides obtained from marine organisms. In particular, it will provide an update on the antiviral actions of the sulfated polysaccharides derived from marine algae including carrageenans, alginates, and fucans, relating to their structure features and the structure–activity relationships. In addition, the recent findings on the different mechanisms of antiviral actions of marine polysaccharides and their potential for therapeutic application will also be summarized in detail.

  20. A simple technique for removing plant polysaccharide contaminants from DNA.

    Science.gov (United States)

    Do, N; Adams, R P

    1991-02-01

    A survey of the inhibitory effects of various plant polysaccharides on DNA restrictions (HindIII and EcoRI) revealed that neutral polysaccharides (arabino-galactan, dextran, gum guar, gum locust bean, beta-glucan, inulin, laminaran, mannan and starch) were not very inhibitory. In contrast, acidic polysaccharides (carrageenan, dextran sulfate, gum ghatti, gum karaya, pectin and xylan) were very inhibitory, even at low concentrations. The Elutip-d (RPC-5 type resin) was evaluated for removal of the inhibitory polysaccharides. Used alone or in combination with a phenol/chloroform wash, it proved effective in removing the polysaccharide so that HindIII digestion was possible, except in the cases of carrageenan and dextran sulfate. In addition, the genomic DNA extracts from live oak (Quercus virginiana) and magnolia (Magnolia grandiflora) were sufficiently purified so that the DNAs could be restricted with both EcoRI and HindIII.

  1. Modified polysaccharides as alternative binders for foundry industry

    Directory of Open Access Journals (Sweden)

    K. Kaczmarska

    2016-10-01

    Full Text Available Polysaccharides constitute a wide group of important polymers with many commercial applications, for example food packaging, fibres, coatings, adhesives etc. This review is devoted to the presentation of polysaccharide application in foundry industry. In this paper the selected properties of foundry moulding sand and core sand containing modified polysaccharides as binders are presented according to foreign literature data. Also, author’s own research about effect of using moulding sand binder consisting of modified polysaccharide (modified starch or its composition with non-toxic synthetic polymers are discussed. Based on technologies taken under consideration in this paper, it could be concluded that polysaccharides are suitable as an alternative for use as binder in foundry moulding applications.

  2. Sulfation of tea polysaccharides: synthesis, characterization and hypoglycemic activity.

    Science.gov (United States)

    Wang, Yuanfeng; Peng, Yonghua; Wei, Xinlin; Yang, Zhiwei; Xiao, Jianbo; Jin, Zhengyu

    2010-03-01

    Neutral polysaccharides (NTPS) and acid polysaccharides (ATPS) from tea leaves were obtained on a D315 macroporous anion-exchange resin column chromatography. NTPS and ATPS were sulfated by the pyridine-sulfonic acid method to obtain NTPS-S and ATPS-S. It was found that NTPS was easier sulfated than ATPS. There are strong characteristic absorption peaks located in 1258 cm(-1), 1146 cm(-1), 832 cm(-1) and 617 cm(-1) in the FTIR spectra of sulfated polysaccharides. Sulfation of polysaccharides also affected the endothermic and exothermic peaks via the DSC scan analysis. The appearance of exothermic peaks in both NTPS-S and ATPS-S indicated that the redox reaction might happen. The comparative study of hypoglycemic effect on mice showed that the sulfation of polysaccharides significantly improved hypoglycemic activity.

  3. The production-influencing factors of extracellular polysacchadde(EPS) from a Strain of lactic acid bacteria and EPS extraction

    Institute of Scientific and Technical Information of China (English)

    CHEN Ying; SUN Liping; ZENG Yong; WANG Lei; AN Liguo

    2006-01-01

    The influencing factors of extracellular polysaccharide(EPS)produced from a strain of lactic acid bacteria(LAB L15)were studied by using the phenol-H2SO4 method.It was demonstrated that the strain produced EPS at the most amount when it was incubated for 40-48 h and when the pH value was 4 under 30℃.Glucose was the most suitable carbon source for LAB-producing EPS.The rough EPS was obtained from L15 culture after centrifugation,dialysis,deprotein,decoloration,and ethanol-precipitation.The sample was at least composed of two polysaccharides mat were completely different in molecular weight and the amount.The purified EPS was passed through the SephadexG-200 colunm and it showed that it was a sample purified by thin layer chromatography.

  4. Overview of microalgal extracellular polymeric substances (EPS) and their applications.

    Science.gov (United States)

    Xiao, Rui; Zheng, Yi

    2016-11-15

    Microalgae have been studied as natural resources for a number of applications, most particularly food, animal feed, biofuels, pharmaceuticals, and nutraceuticals. In addition to the intracellular compounds of interest, microalgae can also excrete various extracellular polymeric substances (EPS) into their immediate living environment during their life cycle to form a hydrated biofilm matrix. These microalgal EPS mainly consist of polysaccharides, proteins, nucleic acids and lipids. Most notably, EPS retain their stable matrix structure and form a 3-D polymer network for cells to interact with each other, and mediate their adhesion to surfaces. EPS also play a role as extracellular energy and carbon sinks. They are also abundant source of structurally and compositionally diverse biopolymers which possess unique bioactivities for special high-value applications, specifically as antivirals, antitumor agents, antioxidants, anticoagulants and anti-inflammatories. Their superior rheological properties also make microalgal EPS particularly useful in mechanical engineering (e.g., biolubricants and drag reducers) and food science/engineering (e.g., thickener and preservatives) applications. The chemical composition and structure of EPS appear to correlate with their applications, but the fundamentals of such relationship are not well understood. This article summarizes previous research on microalgal EPS derived from green algae, diatoms and red algae, including compositions/functions/structure, production, and potential applications. The importance of exopolysaccharides and EPS proteins, with their particular metabolic characteristics, are also described because of their potential high-value applications. This review concludes with potential future research areas of microalgal EPS. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Methods for understanding microbial community structures and functions in microbial fuel cells: a review.

    Science.gov (United States)

    Zhi, Wei; Ge, Zheng; He, Zhen; Zhang, Husen

    2014-11-01

    Microbial fuel cells (MFCs) employ microorganisms to recover electric energy from organic matter. However, fundamental knowledge of electrochemically active bacteria is still required to maximize MFCs power output for practical applications. This review presents microbiological and electrochemical techniques to help researchers choose the appropriate methods for the MFCs study. Pre-genomic and genomic techniques such as 16S rRNA based phylogeny and metagenomics have provided important information in the structure and genetic potential of electrode-colonizing microbial communities. Post-genomic techniques such as metatranscriptomics allow functional characterizations of electrode biofilm communities by quantifying gene expression levels. Isotope-assisted phylogenetic analysis can further link taxonomic information to microbial metabolisms. A combination of electrochemical, phylogenetic, metagenomic, and post-metagenomic techniques offers opportunities to a better understanding of the extracellular electron transfer process, which in turn can lead to process optimization for power output.

  6. Research of polysaccharide complexes from asteraceae family plants

    Directory of Open Access Journals (Sweden)

    Світлана Михайлівна Марчишин

    2015-10-01

    Full Text Available Aim of research. Depth study of polysaccharides in some little-known plant species of Asteraceae family is pressing question, considering that polysaccharides are important biologically active compounds widely used in pharmaceutical and medical practice as remedies and preventive medications. The aim of research was to determinate both quantitative content and monomeric composition of polysaccharide complexes from Asteraceae family plant species – Tagetes genus, Arnica genus, and Bellis genus.Materials and methods. Determination of polysaccharides was carried out by the precipitation reaction, using 96 % ethyl alcohol P and Fehling's solution after acid hydrolysis; quantitative content of this group of compounds was determined by gravimetric analysis. On purpose to identify the monomeric composition hydrolysis under sulfuric acid conditions was conducted. Qualitative monomeric composition of polysaccharides after hydrolysis was carried out by paper chromatography method in n-Butanol – Pyridine – Distilled water P (6:4:3 system along with saccharides reference samples.Results. Polysaccharide complexes from Tagetes erecta, Tagetes patula, Tagetes tenuifolia, Arnica montana, Arnica foliosa, wild and cultivated Bellis perennis herbs were studied. Water-soluble polysaccharides and pectin fractions were isolated from studied objects; their quantitative content and monomeric composition were determined.Conclusion. The highest amount of water-soluble polysaccharides was found in cultivated Bellis perennis herb (10,13 %, the highest amount of pectin compounds – in Tagetes tenuifolia herb (13,62 %; the lowest amount of water-soluble polysaccharides and pectin compounds was found in Arnica montana herb (4,61 % and Tagetes patula herb (3,62 %, respectively. It was found that polysaccharide complexes from all studied species include glucose and arabinose

  7. Atmospheric CO2 and soil extracellular enzyme activity: A meta-analysis and CO2 gradient experiment

    Science.gov (United States)

    Rising atmospheric CO2 concentrations may alter carbon and nutrient cycling and microbial processes in terrestrial ecosystems. One of the primary ways that microbes interact with soil organic matter is through the production of extracellular enzymes, which break down large, complex organic molecules...

  8. A bioelectrochemical approach to characterize extracellular electron transfer by Synechocystis sp. PCC6803.

    Directory of Open Access Journals (Sweden)

    Angelo Cereda

    Full Text Available Biophotovoltaic devices employ photosynthetic organisms at the anode of a microbial fuel cell to generate electrical power. Although a range of cyanobacteria and algae have been shown to generate photocurrent in devices of a multitude of architectures, mechanistic understanding of extracellular electron transfer by phototrophs remains minimal. Here we describe a mediatorless bioelectrochemical device to measure the electrogenic output of a planktonically grown cyanobacterium, Synechocystis sp. PCC6803. Light dependent production of current is measured, and its magnitude is shown to scale with microbial cell concentration and light intensity. Bioelectrochemical characterization of a Synechocystis mutant lacking Photosystem II demonstrates conclusively that production of the majority of photocurrent requires a functional water splitting aparatus and electrons are likely ultimately derived from water. This shows the potential of the device to rapidly and quantitatively characterize photocurrent production by genetically modified strains, an approach that can be used in future studies to delineate the mechanisms of cyanobacterial extracellular electron transport.

  9. Functional polysaccharides as edible coatings for cheese.

    Science.gov (United States)

    Cerqueira, Miguel A; Lima, Alvaro M; Souza, Bartolomeu W S; Teixeira, José A; Moreira, Renato A; Vicente, António A

    2009-02-25

    The objective of the present study was to apply the polysaccharides from different nontraditional sources for cheese coatings. Chitosan, galactomannan from Gleditsia triacanthos, and agar from Glacilaria birdiae were tested, with different formulations and with the addition of plasticizer and corn oil. The surface properties of the cheese and the wetting capacity of the coatings on the cheese were determined. The three best solutions for each polysaccharide were chosen, further films were cast, and permeability to water vapor, oxygen, and carbon dioxide was determined, along with opacity. The solutions of G. triacanthos (formulation: 1.5% of galactomannan, 2.0% of glycerol, and 0.5% of oil) presented the best properties to coat the cheese: -38.76 mN x m(-1) for wettability; 3.24 x 10(-11) (g x (m x s x Pa)(-1)) for water vapor permeability; 0.94 x 10(-15) and 15.35 x 10(-15) (g x m(Pa x s x m(2))(-1)) for oxygen and carbon dioxide permeabilities, respectively; and opacity values of 5.27%. The O(2) consumption and CO(2) production rates of the cheese with and without coating were evaluated, showing a decrease of the respiration rates when the coating was applied. The uncoated cheese had an extensive mold growth at the surface when compared with the coated cheese. The results show that these coatings can be applied as an alternative to synthetic coatings.

  10. Polysaccharide based edible coating on sapota fruit

    Science.gov (United States)

    Menezes, Joslin; Athmaselvi, K. A.

    2016-10-01

    Sapota fruits are highly perishable and have short shelf life at the ambient conditions. The edible coatings have been used on different agricultural products in order to extend their post harvest life. In the present study, the polysaccharide based edible coating made up of sodium alginate and pectin (2%) was studied on the shelf life of sapota fruits. The coating of the fruits is done by dipping method with two dipping time (2 and 4 min). The both control and coated sapota fruits were stored at refrigerated temperature (4±1°C). The physico-chemical analysis including acidity, total soluble solids, ascorbic acid, pH, weight loss, colour and firmness were measured on 1, 8, 15, 23 and 30th day of storage. There was significant difference (p≤0.05) in these physico-chemical parameters between control and coated sapota fruits with 2 and 4 min dipping time. The sensory analysis of control and coated sapota fruits showed that, the polysaccharide coating with 2 minutes dipping time was effective in maintaining the organoleptic properties of the fruits.

  11. Microanalysis of Plant Cell Wall Polysaccharides

    Institute of Scientific and Technical Information of China (English)

    Nicolai Obel; Veronika Erben; Tatjana Schwarz; Stefan Kühne; Andrea Fodor; Markus Pauly

    2009-01-01

    Oligosaccharide Mass Profiling (OLIMP) allows a fast and sensitive assessment of cell wall polymer structure when coupled with Matrix Assisted Laser Desorption Ionisation Time Of Flight Mass Spectrometry (MALDI-TOF MS). The short time required for sample preparation and analysis makes possible the study of a wide range of plant organs, revealing a high degree of heterogeneity in the substitution pattern of wall polymers such as the cross-linking glycan xyloglucan and the pectic polysaccharide homogalacturonan. The high sensitivity of MALDI-TOF allows the use of small amounts of samples, thus making it possible to investigate the wall structure of single cell types when material is collected by such methods as laser micro-dissection. As an example, the analysis of the xyloglucan structure in the leaf cell types outer epidermis layer, entire epidermis cell layer, palisade mesophyll cells, and vascular bundles were investigated. OLIMP is amenable to in situ wall analysis, where wall polymers are analyzed on unprepared plant tissue itself without first iso-lating cell walls. In addition, OLIMP enables analysis of wall polymers in Golgi-enriched fractions, the location of nascent matrix polysaccharide biosynthesis, enabling separation of the processes of wall biosynthesis versus post-deposition apo-plastic metabolism. These new tools will make possible a semi-quantitative analysis of the cell wall at an unprecedented level.

  12. Primary structure and configuration of tea polysaccharide

    Institute of Scientific and Technical Information of China (English)

    ZHOU Peng; XIE Mingyong; NIE Shaoping; WANG Xiaoru

    2004-01-01

    The monosaccharide composition of a tea polysaccharide(TGC)was determined by GC-MS method.Furthermore,the primary structure of tea polysaccharide and its configuration in the aqueous solution were investigated utilizing a combination of classical chemical methods and modern instrumental techniques including GC-MS,Proton NMR,UV and CD.The results indicate that TGC consists of 6 monosaccharides: Rha,Ara,Xyl,Glu,Man and Gal.The configuration of TGC in water solution is proposed to be an ordered helix.The possible primary structure of TGC was outlined as below: the basic structure of the main chain consists of Rha,Glu and Gal units.All three monosaccharides can potentially be connected to branch chains consisting of mainly Ara,and the linkages could be in β1 →2,β1 →3,β2→3 forms.When branch chain is absent in the basic structure of the main chain the linkage consists of only β1→3; Xyl exists at the terminal end of either the main chain or the branch chain with β1 → linkage.

  13. Evaluation of the physico-chemical properties of a new polysaccharide gum from Prosopis africana.

    Science.gov (United States)

    Adikwu, M U; Ezeabasili, S I; Esimone, C O

    2001-01-01

    The gum obtained from the ripe seeds of Prosopis africana was processed to compendial standard for plant gums and characterised. Toxicological studies of the polysaccharide on mice showed the material to be safe. The material hydrates slowly in aqueous media to form a colloidal dispersion. Swelling studies on the gum shows that the gum has a higher swelling capacity than methylcellulose. Rheological studies showed that the material is more viscous than tragacanth gum at equivalent concentrations. Acid hydrolysis and thin layer chromatography of the resulting hydrolysates showed that the gum contains glucose, fructose, galactose and xylose as the monosaccharide components. Microbial tests showed the gum to contain 8.26 x 10(4) viable cells per gram when freshly prepared. Other properties of the gum evaluated includes; melting or charring temperature, optical properties, true density, ash values, element content as well as its reactions with lead subacetate solution and 0.02 M iodine.

  14. Biofilm-specific extracellular matrix proteins of nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Wu, Siva; Baum, Marc M; Kerwin, James; Guerrero, Debbie; Webster, Simon; Schaudinn, Christoph; VanderVelde, David; Webster, Paul

    2014-12-01

    Nontypeable Haemophilus influenzae (NTHi), a human respiratory tract pathogen, can form colony biofilms in vitro. Bacterial cells and the amorphous extracellular matrix (ECM) constituting the biofilm can be separated using sonication. The ECM from 24- and 96-h NTHi biofilms contained polysaccharides and proteinaceous components as detected by nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopy (FTIR) spectroscopy. More conventional chemical assays on the biofilm ECM confirmed the presence of these components and also DNA. Proteomics revealed eighteen proteins present in biofilm ECM that were not detected in planktonic bacteria. One ECM protein was unique to 24-h biofilms, two were found only in 96-h biofilms, and fifteen were present in the ECM of both 24- and 96-h NTHi biofilms. All proteins identified were either associated with bacterial membranes or cytoplasmic proteins. Immunocytochemistry showed two of the identified proteins, a DNA-directed RNA polymerase and the outer membrane protein OMP P2, associated with bacteria and biofilm ECM. Identification of biofilm-specific proteins present in immature biofilms is an important step in understanding the in vitro process of NTHi biofilm formation. The presence of a cytoplasmic protein and a membrane protein in the biofilm ECM of immature NTHi biofilms suggests that bacterial cell lysis may be a feature of early biofilm formation.

  15. Extracellular nucleotide signaling in plants

    Energy Technology Data Exchange (ETDEWEB)

    Stacey, Gary [Univ. of Missouri, Columbia, MO (United States)

    2016-09-08

    Over the life of this funded project, our research group identified and characterized two key receptor proteins in plants; one mediating the innate immunity response to chitin and the other elucidating the key receptor for extracellular ATP. In the case of chitin recognition, we recently described the quaternary structure of this receptor, shedding light on how the receptor functions. Perhaps more importantly, we demonstrated that all plants have the ability to recognize both chitin oligomers and lipochitooligosacchardes, fundamentally changing how the community views the evolution of these systems and strategies that might be used, for example, to extend symbiotic nitrogen fixation to non-legumes. Our discovery of DORN1 opens a new chapter in plant physiology documenting conclusively that eATP is an important extracellular signal in plants, as it is in animals. At this point, we cannot predict just how far reaching this discovery may prove to be but we are convinced that eATP signaling is fundamental to plant growth and development and, hence, we believe that the future will be very exciting for the study of DORN1 and its overall function in plants.

  16. The Role of Shewanella oneidensis MR-1 Outer Surface Structures in Extracellular Electron Transfer

    Science.gov (United States)

    2010-01-01

    bacteria such as Shewanella putrefaciens sp200, Geobacter metallireducens, and G. sulfurreducens [14 – 17]. In several of these bacteria, target- ing of c...Full Paper The Role of Shewanella oneidensis MR-1 Outer Surface Structures in Extracellular Electron Transfer Rachida A. Bouhenni,a, f Gary J. Vora,b...metal reducer Shewanella oneidensis MR-1 to generate electricity in microbial fuel cells (MFCs) depends on the activity of a predicted type IV prepilin

  17. DNase expression allows the pathogen group A streptococcus to escape killing in neutrophil extracellular traps

    OpenAIRE

    Buchanan, John T; Simpson, Amelia J; Aziz, Ramy K.; Liu, George Y.; Kristian, Sascha A.; Kotb, Malak; Feramisco, James; Nizet, Victor

    2006-01-01

    The innate immune response plays a crucial role in satisfactory host resolution of bacterial infection. In response to chemotactic signals, neutrophils are early responding cells that migrate in large numbers to sites of infection. The recent discovery of secreted neutrophil extracellular traps (NETs) composed of DNA and histones opened a novel dimension in our understanding of the microbial killing capacity of these specialized leukocytes. M1 serotype strains of the pathogen Group A Streptoc...

  18. Polysaccharide-Based Nanobiomaterials as Controlled Release Systems for Tissue Engineering Applications.

    Science.gov (United States)

    Rodriguez-Velazquez, Eustolia; Alatorre-Meda, Manuel; Mano, Joao F

    2015-01-01

    Polysaccharides belong to a special class of biopolymers that has been used in different areas of research and technology for some years now. They present distinctive features attractive for the biomedical field. Among others, as extracted from natural sources, these materials are usually biocompatible and possess a significant ability to absorb water. Moreover, they can be conveniently modified by chemical means so as to display improved biological and physicochemical properties. The last but not the least, they are abundant in the natural Extracellular Matrix (ECM) and have a tremendous affinity for different endogenous macromolecules. Accordingly, these particular materials constitute outstanding candidates for a variety of biomimetic approaches entailing the entrapment/stabilization of bioactive molecules (e.g. growth factors, siRNA, and DNA) that could be delivered and have an effect on relevant cellular mechanisms, such as gene expression and cell viability, -proliferation, and -differentiation. This review will explore the current status of nano-scale drug delivery devices based on polysaccharides that could be used in tissue engineering and regenerative medicine (TERM). Aiming to contextualize the topics here discussed, especially for non-experts in the field, section 1 (Introduction) will present a brief overview of TERM and the principal polysaccharides herein employed. In order to get a broader perspective on both issues, this section will include a brief description of non-nanometric systems with relevant characteristics for TERM, such as injectable microparticles and macroscopic hydrogels, just to cite a few. Section 2 will illustrate the contributions of nanotechnology to the development of TERM, in particular to the development of biomimetic systems capable of replicating the natural, endogenous ECMs. Next, sections 3 to 6 will describe representative systems in the nanometric scale presenting 0D (nanoparticles), 1D (nanorods and nanowires), 2D (thin

  19. Microbial conversions of terpenoids

    OpenAIRE

    Parshikov, Igor A

    2015-01-01

    The monograph describes examples of the application of microbial technology for obtaining of derivatives of terpenoids. Obtaining new derivatives of terpenoids, including artemisinin derivatives with increased antimalarial activity, is an important goal of research in microbial biotechnology and medicinal chemistry.

  20. Pseudomonas aeruginosa extracellular products inhibit staphylococcal growth, and disrupt established biofilms produced by Staphylococcus epidermidis

    DEFF Research Database (Denmark)

    Qin, Zhiqiang; Yang, Liang; Qu, Di

    2009-01-01

    Multiple bacterial species often coexist as communities, and compete for environmental resources. Here, we describe how an opportunistic pathogen, Pseudomonas aeruginosa, uses extracellular products to interact with the nosocomial pathogen Staphylococcus epidermidis. S. epidermidis biofilms...... and planktonic cultures were challenged with P. aeruginosa supernatant cultures overnight. Results indicated that quorum-sensing-controlled factors from P. aeruginosa supernatant inhibited S. epidermidis growth in planktonic cultures. We also found that P. aeruginosa extracellular products, mainly...... in overnight cultures had no effect on established P. aeruginosa biofilms and planktonic growth. These findings reveal that P. aeruginosa extracellular products are important microbial competition factors that overcome competition with S. epidermidis, and the results may provide clues for the development...

  1. Antibiofilm Activity of Actinobacillus pleuropneumoniae Serotype 5 Capsular Polysaccharide

    Science.gov (United States)

    Karwacki, Michael T.; Kadouri, Daniel E.; Bendaoud, Meriem; Izano, Era A.; Sampathkumar, Vandana; Inzana, Thomas J.; Kaplan, Jeffrey B.

    2013-01-01

    Cell-free extracts isolated from colony biofilms of Actinobacillus pleuropneumoniae serotype 5 were found to inhibit biofilm formation by Staphylococcus aureus, S. epidermidis and Aggregatibacter actinomycetemcomitans, but not by A. pleuropneumoniae serotype 5 itself, in a 96-well microtiter plate assay. Physical and chemical analyses indicated that the antibiofilm activity in the extract was due to high-molecular-weight polysaccharide. Extracts isolated from a mutant strain deficient in the production of serotype 5 capsular polysaccharide did not exhibit antibiofilm activity. A plasmid harboring the serotype 5 capsule genes restored the antibiofilm activity in the mutant extract. Purified serotype 5 capsular polysaccharide also exhibited antibiofilm activity against S. aureus. A. pleuropneumoniae wild-type extracts did not inhibit S. aureus growth, but did inhibit S. aureus intercellular adhesion and binding of S. aureus cells to stainless steel surfaces. Furthermore, polystyrene surfaces coated with A. pleuropneumoniae wild-type extracts, but not with capsule-mutant extracts, resisted S. aureus biofilm formation. Our findings suggest that the A. pleuropneumoniae serotype 5 capsule inhibits cell-to-cell and cell-to-surface interactions of other bacteria. A. pleuropneumoniae serotype 5 capsular polysaccharide is one of a growing number of bacterial polysaccharides that exhibit broad-spectrum, nonbiocidal antibiofilm activity. Future studies on these antibiofilm polysaccharides may uncover novel functions for bacterial polysaccharides in nature, and may lead to the development of new classes of antibiofilm agents for industrial and clinical applications. PMID:23691104

  2. Antibiofilm activity of Actinobacillus pleuropneumoniae serotype 5 capsular polysaccharide.

    Directory of Open Access Journals (Sweden)

    Michael T Karwacki

    Full Text Available Cell-free extracts isolated from colony biofilms of Actinobacillus pleuropneumoniae serotype 5 were found to inhibit biofilm formation by Staphylococcus aureus, S. epidermidis and Aggregatibacter actinomycetemcomitans, but not by A. pleuropneumoniae serotype 5 itself, in a 96-well microtiter plate assay. Physical and chemical analyses indicated that the antibiofilm activity in the extract was due to high-molecular-weight polysaccharide. Extracts isolated from a mutant strain deficient in the production of serotype 5 capsular polysaccharide did not exhibit antibiofilm activity. A plasmid harboring the serotype 5 capsule genes restored the antibiofilm activity in the mutant extract. Purified serotype 5 capsular polysaccharide also exhibited antibiofilm activity against S. aureus. A. pleuropneumoniae wild-type extracts did not inhibit S. aureus growth, but did inhibit S. aureus intercellular adhesion and binding of S. aureus cells to stainless steel surfaces. Furthermore, polystyrene surfaces coated with A. pleuropneumoniae wild-type extracts, but not with capsule-mutant extracts, resisted S. aureus biofilm formation. Our findings suggest that the A. pleuropneumoniae serotype 5 capsule inhibits cell-to-cell and cell-to-surface interactions of other bacteria. A. pleuropneumoniae serotype 5 capsular polysaccharide is one of a growing number of bacterial polysaccharides that exhibit broad-spectrum, nonbiocidal antibiofilm activity. Future studies on these antibiofilm polysaccharides may uncover novel functions for bacterial polysaccharides in nature, and may lead to the development of new classes of antibiofilm agents for industrial and clinical applications.

  3. The immunostimulating role of lichen polysaccharides: a review.

    Science.gov (United States)

    Shrestha, Gajendra; St Clair, Larry L; O'Neill, Kim L

    2015-03-01

    The immune system has capacity to suppress the development or progression of various malignancies including cancer. Research on the immunomodulating properties of polysaccharides obtained from plants, microorganisms, marine organisms, and fungi is growing rapidly. Among the various potential sources, lichens, symbiotic systems involving a fungus and an alga and/or a cyanobacterium, show promise as a potential source of immunomodulating compounds. It is well known that lichens produce an abundance of structurally diverse polysaccharides. However, only a limited number of studies have explored the immunostimulating properties of lichen polysaccharides. Published studies have shown that some lichen polysaccharides enhance production of nitrous oxide (NO) by macrophages and also alter the production levels of various proinflammatory and antiinflammatory cytokines (IL-10, IL-12, IL-1β, TNF-α, and IFN-α/β) by macrophages and dendritic cells. Although there are only a limited number of studies examining the role of lichen polysaccharides, all results suggest that lichen polysaccharides can induce immunomodulatory responses in macrophages and dendritic cells. Thus, a detailed evaluation of immunomodulatory capacity of lichen polysaccharides could provide a unique opportunity for the discovery of novel therapeutic agents.

  4. New polysaccharide-based polymer electrolytes; Nouveaux electrolytes polymeres a base de polysaccharides

    Energy Technology Data Exchange (ETDEWEB)

    Velasquez-Morales, P.; Le Nest, J.F.; Gandini, A. [Ecole Francaise de Papeterie et des Industries Graphique, 38 - Saint Martin d`Heres (France)

    1996-12-31

    Polysaccharides like cellulose and chitosan are known for their filmic properties. This paper concerns the synthesis and the study of chitosan-based polymer electrolytes. A preliminary work concerns the study of glucosamine reactivity. The poly-condensation of chitosan ethers (obtained by reaction with ethylene oxide or propylene oxide) with bifunctional and monofunctional oligo-ethers leads to the formation of thin lattices (10 {mu}m) having excellent mechanical properties. The presence of grafted polyether chains along the polysaccharide skeleton allows to modify the vitreous transition temperature and the molecular disorder of the system. Two type of polymer electrolytes have been synthesized: electrolytes carrying a dissolved alkaline metal salt and ionomers. The analysis of their thermal, dynamical mechanical, nuclear magnetic relaxation, electrical, and electrochemical properties shows that this new class of polymer electrolytes has the same performances as ethylene poly-oxide based amorphous lattices plus the advantage of having good filmic properties. Abstract only. (J.S.)

  5. Microfluidics and microbial engineering.

    Science.gov (United States)

    Kou, Songzi; Cheng, Danhui; Sun, Fei; Hsing, I-Ming

    2016-02-01

    The combination of microbial engineering and microfluidics is synergistic in nature. For example, microfluidics is benefiting from the outcome of microbial engineering and many reported point-of-care microfluidic devices employ engineered microbes as functional parts for the microsystems. In addition, microbial engineering is facilitated by various microfluidic techniques, due to their inherent strength in high-throughput screening and miniaturization. In this review article, we firstly examine the applications of engineered microbes for toxicity detection, biosensing, and motion generation in microfluidic platforms. Secondly, we look into how microfluidic technologies facilitate the upstream and downstream processes of microbial engineering, including DNA recombination, transformation, target microbe selection, mutant characterization, and microbial function analysis. Thirdly, we highlight an emerging concept in microbial engineering, namely, microbial consortium engineering, where the behavior of a multicultural microbial community rather than that of a single cell/species is delineated. Integrating the disciplines of microfluidics and microbial engineering opens up many new opportunities, for example in diagnostics, engineering of microbial motors, development of portable devices for genetics, high throughput characterization of genetic mutants, isolation and identification of rare/unculturable microbial species, single-cell analysis with high spatio-temporal resolution, and exploration of natural microbial communities.

  6. Childhood microbial keratitis

    Directory of Open Access Journals (Sweden)

    Abdullah G Al Otaibi

    2012-01-01

    Conclusion: Children with suspected microbial keratitis require comprehensive evaluation and management. Early recognition, identifying the predisposing factors and etiological microbial organisms, and instituting appropriate treatment measures have a crucial role in outcome. Ocular trauma was the leading cause of childhood microbial keratitis in our study.

  7. Redox properties of extracellular polymeric substances (EPS) from electroactive bacteria

    Science.gov (United States)

    Li, Shan-Wei; Sheng, Guo-Ping; Cheng, Yuan-Yuan; Yu, Han-Qing

    2016-12-01

    Although the capacity for electroactive bacteria to convert environmental metallic minerals and organic pollutants is well known, the role of the redox properties of microbial extracellular polymeric substances (EPS) in this process is poorly understood. In this work, the redox properties of EPS from two widely present electroactive bacterial strains (Shewanella oneidensis and Pseudomonas putida) were explored. Electrochemical analysis demonstrates that the EPS extracted from the two strains exhibited redox properties. Spectroelectrochemical and protein electrophoresis analyses indicate that the extracted EPS from S. oneidensis and P. putida contained heme-binding proteins, which were identified as the possible redox components in the EPS. The results of heme-mediated behavior of EPS may provide an insight into the important roles of EPS in electroactive bacteria to maximize their redox capability for biogeochemical cycling, environmental bioremediation and wastewater treatment.

  8. Analysis of extracellular RNA by digital PCR

    Directory of Open Access Journals (Sweden)

    Kenji eTakahashi

    2014-06-01

    Full Text Available The transfer of extracellular RNA is emerging as an important mechanism for intracellular communication. The ability for the transfer of functionally active RNA molecules from one cell to another within vesicles such as exosomes enables a cell to modulate cellular signaling and biological processes within recipient cells. The study of extracellular RNA requires sensitive methods for the detection of these molecules. In this methods article, we will describe protocols for the detection of such extracellular RNA using sensitive detection technologies such as digital PCR. These protocols should be valuable to researchers interested in the role and contribution of extracellular RNA to tumor cell biology.

  9. Extracellular Molecules Involved in Cancer Cell Invasion

    Energy Technology Data Exchange (ETDEWEB)

    Stivarou, Theodora; Patsavoudi, Evangelia, E-mail: epatsavoudi@pasteur.gr [Department of Biochemistry, Hellenic Pasteur Institute, Athens 11521 (Greece); Technological Educational Institute of Athens, Egaleo, Athens 12210 (Greece)

    2015-01-26

    Nowadays it is perfectly clear that understanding and eradicating cancer cell invasion and metastasis represent the crucial, definitive points in cancer therapeutics. During the last two decades there has been a great interest in the understanding of the extracellular molecular mechanisms involved in cancer cell invasion. In this review, we highlight the findings concerning these processes, focusing in particular on extracellular molecules, including extracellular matrix proteins and their receptors, growth factors and their receptors, matrix metalloproteinases and extracellular chaperones. We report the molecular mechanisms underlying the important contribution of this pool of molecules to the complex, multi-step phenomenon of cancer cell invasion.

  10. Extracellular Molecules Involved in Cancer Cell Invasion

    Directory of Open Access Journals (Sweden)

    Theodora Stivarou

    2015-01-01

    Full Text Available Nowadays it is perfectly clear that understanding and eradicating cancer cell invasion and metastasis represent the crucial, definitive points in cancer therapeutics. During the last two decades there has been a great interest in the understanding of the extracellular molecular mechanisms involved in cancer cell invasion. In this review, we highlight the findings concerning these processes, focusing in particular on extracellular molecules, including extracellular matrix proteins and their receptors, growth factors and their receptors, matrix metalloproteinases and extracellular chaperones. We report the molecular mechanisms underlying the important contribution of this pool of molecules to the complex, multi-step phenomenon of cancer cell invasion.

  11. Chemical Structures and Bioactivities of Sulfated Polysaccharides from Marine Algae

    Directory of Open Access Journals (Sweden)

    H. Stephen Ewart

    2011-02-01

    Full Text Available Sulfated polysaccharides and their lower molecular weight oligosaccharide derivatives from marine macroalgae have been shown to possess a variety of biological activities. The present paper will review the recent progress in research on the structural chemistry and the bioactivities of these marine algal biomaterials. In particular, it will provide an update on the structural chemistry of the major sulfated polysaccharides synthesized by seaweeds including the galactans (e.g., agarans and carrageenans, ulvans, and fucans. It will then review the recent findings on the anticoagulant/antithrombotic, antiviral, immuno-inflammatory, antilipidemic and antioxidant activities of sulfated polysaccharides and their potential for therapeutic application.

  12. Composite polysaccharide fibers prepared by electrospinning and coating.

    Science.gov (United States)

    Maeda, N; Miao, J; Simmons, T J; Dordick, J S; Linhardt, R J

    2014-02-15

    Composite polysaccharide fibers composed two oppositely charged natural polysaccharides, chitosan and hyaluronic acid, were prepared by electrospinning and subsequent coating. The fiber size distribution was characterized by scanning electron microscopy. Chitosan/hyaluronic acid composite fibers were stable in water but showed controlled release of hyaluronic acid into phosphate buffered saline, and the presence of 3-wt% hyaluronic acid coating improved the swelling ratio to 30%. The resulting composite polysaccharide fibers have a number of potential biomedical applications in wound healing applications and in drug delivery systems.

  13. Navicula sp. Sulfated Polysaccharide Gels Induced by Fe(III): Rheology and Microstructure

    OpenAIRE

    2016-01-01

    A sulfated polysaccharide extracted from Navicula sp. presented a yield of 4.4 (% w/w dry biomass basis). Analysis of the polysaccharide using gas chromatography showed that this polysaccharide contained glucose (29%), galactose (21%), rhamnose (10%), xylose (5%) and mannose (4%). This polysaccharide presented an average molecular weight of 107 kDa. Scanning electron microscopy (SEM) micrographs showed that the lyophilized Navicula sp. polysaccharide is an amorphous solid with particles of ir...

  14. Polysaccharide Responsiveness Is Not Biased by Prior Pneumococcal-Conjugate Vaccination

    Science.gov (United States)

    Bernth-Jensen, Jens Magnus; Søgaard, Ole Schmeltz

    2013-01-01

    Polysaccharide responsiveness is tested by measuring antibody responses to polysaccharide vaccines to diagnose for humoral immunodeficiency. A common assumption is that this responsiveness is biased by any previous exposure to the polysaccharides in the form of protein-coupled polysaccharide vaccines, such as those used in many childhood vaccination programmes. To examine this assumption, we investigated the effect of protein-coupled polysaccharide vaccination on subsequent polysaccharide responsiveness. HIV-infected adults (n = 47) were vaccinated twice with protein-coupled polysaccharides and six months later with pure polysaccharides. We measured immunoglobulin G responses against three polysaccharides present in only the polysaccharide vaccine (non-memory polysaccharides) and seven recurring polysaccharides (memory polysaccharides). Responsiveness was evaluated according to the consensus guidelines published by the American immunology societies. Impaired responsiveness to non-memory polysaccharides was more frequent than to memory polysaccharides (51% versus 28%, P = 0.015), but the individual polysaccharides did not differ in triggering sufficient responses (74% versus 77%, P = 0.53). Closer analysis revealed important shortcomings of the current evaluation guidelines. The interpreted responseś number and their specificities influenced the likelihood of impaired responsiveness in a complex manor. This influence was propelled by the dichotomous approaches inherent to the American guidelines. We therefore define a novel more robust polysaccharide responsiveness measure, the Z-score, which condenses multiple, uniformly weighted responses into one continuous variable. Using the Z-score, responsiveness to non-memory polysaccharides and memory-polysaccharides were found to correlate (R2 = 0.59, Presponsiveness was not biased by prior protein-coupled polysaccharide vaccination in HIV-infected adults. Studies in additional populations are warranted

  15. An Acidic Polysaccharide from Tribulus terrestris

    Institute of Scientific and Technical Information of China (English)

    HaiShengCHEN; WingNangLEUNG; 等

    2002-01-01

    An aqueous acidic polysaccharide, named rhamnogalacturonan (designated as TIP-D2) was isolated from Tribulus terrestris L by means of DEAE-cellulose chromatography and gel filtration. The molecular mass of TTP-D2 was estimated to be 26 KDa by gel filtration.TTP-D2 is composed of galacturonic acid, rhamnose, arabinose, galactose,fucose,mannose,xylose and glucose in a ratio of 71.4:13.5:5.6:4.9:3.1:1.9:1.9:1.0. The main chain structure of TTP-D2 was elucidated as an acidic hetero-polysaccaride with the connection of α-(1-4) galacturonic acid with α-(1-3) rhamnose by GC analysis of partially hydrolyzed products and determination of 1H,13C-NMR spectra.

  16. An Acidic Polysaccharide from Tribulus terrestris

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    An aqucous acidic polysaccharide, named rhamnogalacturonan (designated as TTP-D2)was isolated from Tribulus terrestris L by means of DEAE-cellulose chromatography and gel filtration. The molecular mass of TTP-D2 was estimated to be 26 KDa by gel filtration. TTP-D2 is composed of galacturonic acid, rhamnose, arabinose, galactose, fucose, mannosc, xylose and glucose in a ratio of 71.4: 13.5: 5.6: 4.9: 3.1: 1.9: 1.9: 1.0. The main chain structure of TTP-D2 was elucidated as an acidic hetero-polysaccharidc with the connection of α-(l-4) galacturonic acid with α-(1-3) rhamnose by GC analysis of partially hydrolyzed products and the determination of 1H, 13C-NMR spectra.

  17. NMR studies of polysaccharides from brown seaweeds

    Energy Technology Data Exchange (ETDEWEB)

    Noseda, M.D.; Tisher, C.A.; Gorin, P.A.J.; Duarte, M.E.R. [Parana Univ., Curitiba, PR (Brazil). Dept. de Bioquimica; Cerezo, A.S. [Buenos Aires Univ. Nacional (Argentina). Dept. de Quimica Organica

    1997-12-31

    Alginic acid is the major intercellular polysaccharide serving as matrix in the brown algae and is comprised of an unbranched chain of (1->4)-linked {beta}-D-mannuronic acid (M) and {alpha}-L-guluronic acid (G), arranged in a blockwise fashion. The composition of the monomer residues and the block structure varies depending on the source of the polymer. The selective binding of cations to alginate accounts for its ability to form gels, which is dependent on the number and lenght of the G-blocks. They are widely used industrially for their ability to retain water, and for their gelling, viscosifying and stabilizing properties (Smidsrod and draget, 1996). In this study, alginate composition and block structure in Sargassum stenophyllum has been determined by chemical methods and NMR spectroscopic analysis. (author) 4 refs., 3 figs.

  18. Use of paramagnetic chelated metal derivatives of polysaccharides and spin-labeled polysaccharides as contrast agents in magnetic resonance imaging

    Energy Technology Data Exchange (ETDEWEB)

    Bligh, S.W.; Harding, C.T.; Sadler, P.J.; Bulman, R.A.; Bydder, G.M.; Pennock, J.M.; Kelly, J.D.; Latham, I.A.; Marriott, J.A. (Department of Chemistry, Birkbeck College, London (England))

    1991-02-01

    Soluble and insoluble polysaccharides were derivatized with diethylenetriaminepentaacetic acid (DTPA) and/or spin-labeled with 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO). Polysaccharides derivatized with DTPA were prepared via cyanogen bromide activation, coupling to a diamine linker, and to DTPA anhydride. Spin-labeled polysaccharides were also prepared via cyanogen bromide activation. The extent of derivatization for dextran (18 kDa) was about 120 glucose units per DTPA, and for cellulose and starch about 15-30 units per DTPA. For spin-labeled polysaccharides, the average loading ranged from 1 nitroxide per 16 glucose units for starch to 181 for dextran (82 kDa). These derivatized paramagnetic polysaccharides were shown to be more effective relaxants than the small paramagnetic molecules alone. Both soluble and insoluble polysaccharide-linker-DTPA-Gd(3) complexes were effectively cleared from the body (rats) after oral administration. After intravenous administration, the biodistribution of dextran-linker-DTPA-Gd(3) complexes differed significantly from that of GdDTPA. Reduction of the nitroxide by ascorbic acid was retarded in the polysaccharide derivatives, particularly in starch derivatized with both nitroxide and linker-DTPA-Cu(2). These agents showed contrast enhancement in the gastrointestinal tract of rabbits.

  19. Utilization of polysaccharides by radiation processing

    Energy Technology Data Exchange (ETDEWEB)

    Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    Radiation treatment has been applied for improvement or pasteurization of agro-resources to recycle the resources and to reduce the pollution of environment. By using the radiation effect for pasteurization, upgrading of cellulosic wastes of oil palm to animal feeds and mushroom has been studied under the bilateral research cooperation between JAERI and MINT (Malaysian Institute for Nuclear Technology Research). The necessary dose for pasteurization of oil palm empty fruit bunch (EFB), which is a main cellulosic by-product of palm oil industry, was determined as 10 kGy. After pasteurization, the EFB substrate was inoculated with Pleurotus sajor-caju and fermented for 1 month. The digestibility and nutritional value of fermented products were evaluated as ruminant feeds and the mushroom can be produced as by-product. For the improvement of resources, radiation effects on polysaccharides such as chitosan, sodium alginate, carrageenan, cellulose, pectin have been investigated to induce the biological activities. These carbohydrates were easily degraded by irradiation and induced various kinds of biological activities. The anti-bacterial activity and elicitor activity of chitosan were induced by irradiation. The induction of phytoalexins was also observed by irradiated pectin but the higher elicitor activity for pisatin was obtained by chitosan than pectin. For the plant growth promotion, alginate derived from brown marine algae, chitosan and ligno-cellulosic extracts show a strong activity. carrageenan derived from red marine algae can promote growth of rice and the highest effect was obtained with kappa carrageenan irradiated at 100 kGy. Furthermore, some radiation degraded polysaccharides suppressed the damage of environmental stress on plants. (author)

  20. Campylobacter polysaccharide capsules: virulence and vaccines

    Directory of Open Access Journals (Sweden)

    Patricia eGuerry

    2012-02-01

    Full Text Available Campylobacter jejuni remains a major cause of bacterial diarrhea worldwide and is associated with numerous sequelae, including Guillain Barre Syndrome, inflammatory bowel disease, reactive arthritis, and irritable bowel syndrome. C. jejuni is unusual for an intestinal pathogen in its ability to coat its surface with a polysaccharide capsule (CPS. These capsular polysaccharides vary in sugar composition and linkage, especially those involving heptoses of unusual configuration and O-methyl phosphoramidate linkages. This structural diversity is consistent with CPS being the major serodeterminant of the Penner scheme, of which there are 47 C. jejuni serotypes. Both CPS expression and expression of modifications are subject to phase variation by slip strand mismatch repair. Although capsules are virulence factors for other pathogens, the role of CPS in C. jejuni disease has not been well defined beyond descriptive studies demonstrating a role in serum resistance and for diarrhea in a ferret model of disease. However, perhaps the most compelling evidence for a role in pathogenesis are data that CPS conjugate vaccines protect against diarrheal disease in non-human primates. A CPS conjugate vaccine approach against this pathogen is intriguing, but several questions need to be addressed, including the valency of CPS types required for an effective vaccine. There have been numerous studies of prevalence of CPS serotypes in the developed world, but few studies from developing countries where the disease incidence is higher. The complexity and cost of Penner serotyping has limited its usefulness, and a recently developed multiplex PCR method for determination of capsule type offers the potential of a more rapid and affordable method. Comparative studies have shown a strong correlation of the two methods and studies are beginning to ascertain CPS-type distribution worldwide, as well as examination of correlation of severity of illness with specific CPS types.

  1. Microbial Enzymatic Degradation of Biodegradable Plastics.

    Science.gov (United States)

    Roohi; Bano, Kulsoom; Kuddus, Mohammed; Zaheer, Mohammed R; Zia, Qamar; Khan, Mohammed F; Ashraf, Ghulam Md; Gupta, Anamika; Aliev, Gjumrakch

    2017-01-01

    The renewable feedstock derived biodegradable plastics are important in various industries such as packaging, agricultural, paper coating, garbage bags and biomedical implants. The increasing water and waste pollution due to the available decomposition methods of plastic degradation have led to the emergence of biodegradable plastics and biological degradation with microbial (bacteria and fungi) extracellular enzymes. The microbes utilize biodegradable polymers as the substrate under starvation and in unavailability of microbial nutrients. Microbial enzymatic degradation is suitable from bioremediation point of view as no waste accumulation occurs. It is important to understand the microbial interaction and mechanism involved in the enzymatic degradation of biodegradable plastics under the influence of several environmental factors such as applied pH, thermo-stability, substrate molecular weight and/or complexity. To study the surface erosion of polymer film is another approach for hydrolytic degradation characteristion. The degradation of biopolymer is associated with the production of low molecular weight monomer and generation of carbon dioxide, methane and water molecule. This review reported the degradation study of various existing biodegradable plastics along with the potent degrading microbes (bacteria and fungi). Patents available on plastic biodegradation with biotechnological significance is also summarized in this paper. This paper assesses that new disposal technique should be adopted for the degradation of polymers and further research is required for the economical production of biodegradable plastics along with their enzymatic degradation. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  2. Detection of Inulin, a Prebiotic Polysaccharide, in Maple Syrup.

    Science.gov (United States)

    Sun, Jiadong; Ma, Hang; Seeram, Navindra P; Rowley, David C

    2016-09-28

    Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. During thermal evaporation of water, natural phytochemical components are concentrated in maple syrup. The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides.

  3. Radiation processed hydrogel of poly (vinyl alcohol) with biodegradable polysaccharides.

    Science.gov (United States)

    Chowdhury, M N K; Alam, A K M M; Dafader, N C; Haque, M E; Akhtar, F; Ahmed, M U; Rashid, H; Begum, R

    2006-01-01

    Poly(vinyl alcohol) (PVA) can be modified to polymer hydrogels by radiation crosslinking and can be used in different biomedical applications. A study was done on the optimization of ingredients concentration for preparing good quality PVA hydrogels with natural polysaccharides. The synthesized hydrogels were also characterized by measuring the different physical properties e.g. gel fraction, swelling and absorption rate. Besides these, sterility test were also performed. Good quality hydrogels were obtained from PVA and natural polysaccharides solutions with 27 kGy radiation dose. There is an influence of natural polysaccharides on the gel fraction of hydrogel. The increase in the amount of polysaccharide causes a decrease in gel fraction that is decrease in the crosslinking density of PVA hydrogel network. The prepared hydrogels were found to be sterile.

  4. Extraction and antioxidant activity of polysaccharides from Rana chensinensis skin.

    Science.gov (United States)

    Wang, Zhanyong; Zhao, Yuanyuan; Su, Tingting

    2015-01-22

    The extraction process of polysaccharides from Rana chensinensis skin was optimized by using a Box-Behnken design. The optimum extraction conditions were as follows: extraction time, 4.96h; extraction temperature, 100°C; ratio of water to raw material, 60; and extraction frequency, 1. Under these conditions, the experimental polysaccharide yield was 2.03±0.14%, which agreed with the predicted yield. The purified polysaccharide RCSP II was successfully obtained by diethylaminoethanol-Sepharose and Sepharose CL-6B column chromatography. In vitro experiments showed that RCSP II exhibited a strong scavenging activity against superoxide anion and 1,1-diphenyl-2-picrylhydrazyl radicals but a weak scavenging activity against hydroxyl radicals. RCSP II also showed a strong reducing capacity. Thus, this polysaccharide can be used as a natural antioxidant in functional foods or medicines.

  5. Degradation studies on Escherichia coli capsular polysaccharides by bacteriophages.

    Science.gov (United States)

    Nimmich, W

    1997-08-01

    The serologically and structurally related Eschrichia coli capsular polysaccharides (K antigens) K13, K20, and K23 were found to be depolymerized by the bacteriophages phi K13 and phi K20 to almost similar oligomer profiles as shown by polyacrylamide gel electrophoresis. The phage-polysaccharide interactions were followed by an increase of reducing 2-keto-3-deoxyoctulosonic acid due to a phage-associated glycanase that catalyzed the hydrolytic cleavage of common beta-ketopyranosidic 2-keto-3-deoxyoctulosonic acid linkages. The related E. coli K antigens K18, K22, and K100 as well as the Haemophilus influenzae type b capsular polysaccharide were degraded by bacteriophage phi K100 with different efficacy. It is suggested that phi K100 enzymatically cleaves ribitol-5-phosphate bonds as the only structural feature present in all the polysaccharides investigated.

  6. Synbiotic matrices derived from plant oligosaccharides and polysaccharides

    Science.gov (United States)

    A porous synbiotic matrix was prepared by lyophilization of alginate and pectin or fructan oligosaccharides and polysaccharides cross-linked with calcium. These synbiotic matrices were excellent physical structures to support the growth of Lactobacillus acidophilus (1426) and Lactobacillus reuteri (...

  7. Comparison of Polysaccharides from Two Species of Ganoderma

    Directory of Open Access Journals (Sweden)

    Yu-Ping Tang

    2012-01-01

    Full Text Available Ganoderma lucidum and Ganoderma sinense, known as Lingzhi in Chinese, are commonly used Chinese medicines with excellent beneficial health effects. Triterpenes and polysaccharides are usually considered as their main active components. However, the content of triterpenes differs significantly between the two species of Ganoderma. To date, a careful comparison of polysaccharides from the two species of Ganoderma has not been performed. In this study, polysaccharides from fruiting bodies of two species of Lingzhi collected from different regions of China were analyzed and compared based on HPSEC-ELSD and HPSEC-MALLS-RI analyses, as well as enzymatic digestion and HPTLC of acid hydrolysates. The results indicated that both the HPSEC-ELSD profiles and the molecular weights of the polysaccharides were similar. Enzymatic digestion showed that polyshaccharides from all samples of Lingzhi could be hydrolyzed by pectinase and dextranase. HPTLC profiles of their TFA hydrolysates colored with different reagents and their monosaccharides composition were also similar.

  8. Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides.

    Science.gov (United States)

    Wasser, S P

    2002-11-01

    The number of mushrooms on Earth is estimated at 140,000, yet maybe only 10% (approximately 14,000 named species) are known. Mushrooms comprise a vast and yet largely untapped source of powerful new pharmaceutical products. In particular, and most importantly for modern medicine, they represent an unlimited source of polysaccharides with antitumor and immunostimulating properties. Many, if not all, Basidiomycetes mushrooms contain biologically active polysaccharides in fruit bodies, cultured mycelium, culture broth. Data on mushroom polysaccharides have been collected from 651 species and 7 infraspecific taxa from 182 genera of higher Hetero- and Homobasidiomycetes. These polysaccharides are of different chemical composition, with most belonging to the group of beta-glucans; these have beta-(1-->3) linkages in the main chain of the glucan and additional beta-(1-->6) branch points that are needed for their antitumor action. High molecular weight glucans appear to be more effective than those of low molecular weight. Chemical modification is often carried out to improve the antitumor activity of polysaccharides and their clinical qualities (mostly water solubility). The main procedures used for chemical improvement are: Smith degradation (oxydo-reducto-hydrolysis), formolysis, and carboxymethylation. Most of the clinical evidence for antitumor activity comes from the commercial polysaccharides lentinan, PSK (krestin), and schizophyllan, but polysaccharides of some other promising medicinal mushroom species also show good results. Their activity is especially beneficial in clinics when used in conjunction with chemotherapy. Mushroom polysaccharides prevent oncogenesis, show direct antitumor activity against various allogeneic and syngeneic tumors, and prevent tumor metastasis. Polysaccharides from mushrooms do not attack cancer cells directly, but produce their antitumor effects by activating different immune responses in the host. The antitumor action of

  9. Engineering microbial electrocatalysis for chemical and fuel production.

    Science.gov (United States)

    Rosenbaum, Miriam A; Henrich, Alexander W

    2014-10-01

    In many biotechnological areas, metabolic engineering and synthetic biology have become core technologies for biocatalyst development. Microbial electrocatalysis for biochemical and fuel production is still in its infancy and reactions rates and the product spectrum are currently very low. Therefore, molecular engineering strategies will be crucial for the advancement and realization of many new bioproduction routes using electroactive microorganisms. The complex and unresolved biochemistry and physiology of extracellular electron transfer and the lack of molecular tools for these new non-model hosts for genetic engineering constitute the major challenges for this effort. This review is providing an insight into the current status, challenges and promising approaches of pathway engineering for microbial electrocatalysis.

  10. Identification of a capsular polysaccharide from Moraxella bovis.

    Science.gov (United States)

    Wilson, Jennifer C; Hitchen, Paul G; Frank, Martin; Peak, Ian R; Collins, Patrick M; Morris, Howard R; Dell, Anne; Grice, I Darren

    2005-03-21

    The bacterium Moraxella bovis is the causative agent of an economically important disease of cattle: Infectious Bovine Keratoconjunctivitis (IBK), otherwise known as pinkeye. Little is known regarding the structure of the carbohydrates produced by M. bovis. The structure of a capsular polysaccharide from M. bovis (strain Mb25) has been determined using NMR and MS analysis. From these data it is concluded that the polysaccharide is composed of the unmodified chondroitin disaccharide repeat unit.

  11. Composite polysaccharide fibers prepared by electrospinning and coating

    OpenAIRE

    2013-01-01

    Composite polysaccharide fibers composed two oppositely charged natural polysaccharides, chitosan and hyaluronic acid, were prepared by electrospinning and subsequent coating The fiber size distribution was characterized by scanning electron microscopy. Chitosan/hyaluronic acid composite fibers were stable in water but showed controlled release of hyaluronic acid into phosphate buffered saline, and the presence of 3-wt% hyaluronic acid coating improved the swelling ratio to 30%. The resulting...

  12. Characterization of Polysaccharide by HPLC: Extraction and Anticancer Effects

    OpenAIRE

    Liming Gao; Ya Di; Jiandong Wu; Ming Shi; Fulu Zheng

    2014-01-01

    Cervical cancer is a serious health hazard for women’s reproductive system cancer; the method of treatment for cervical cancer is still in surgery, chemotherapy, and radiotherapy as the basic means, but with many complications. The effects of natural medicines for cervical cancer are increasingly becoming the focus of people’s attentions. By studying the polysaccharide of cervical cancer in mice, we found that shark cartilage polysaccharide can increase the serum levels of T-SOD and GSH and d...

  13. Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps.

    Science.gov (United States)

    Ávila, Eva E; Salaiza, Norma; Pulido, Julieta; Rodríguez, Mayra C; Díaz-Godínez, César; Laclette, Juan P; Becker, Ingeborg; Carrero, Julio C

    2016-01-01

    Neutrophil defense mechanisms include phagocytosis, degranulation and the formation of extracellular traps (NET). These networks of DNA are triggered by several immune and microbial factors, representing a defense strategy to prevent microbial spread by trapping/killing pathogens. This may be important against Entamoeba histolytica, since its large size hinders its phagocytosis. The aim of this study was to determine whether E. histolytica and their lipopeptidophosphoglycan (EhLPPG) induce the formation of NETs and the outcome of their interaction with the parasite. Our data show that live amoebae and EhLPPG, but not fixed trophozoites, induced NET formation in a time and dose dependent manner, starting at 5 min of co-incubation. Although immunofluorescence studies showed that the NETs contain cathelicidin LL-37 in close proximity to amoebae, the trophozoite growth was only affected when ethylene glycol tetra-acetic acid (EGTA) was present during contact with NETs, suggesting that the activity of enzymes requiring calcium, such as DNases, may be important for amoeba survival. In conclusion, E. histolytica trophozoites and EhLPPG induce in vitro formation of human NETs, which did not affect the parasite growth unless a chelating agent was present. These results suggest that NETs may be an important factor of the innate immune response during infection with E. histolytica.

  14. Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps

    Science.gov (United States)

    Ávila, Eva E.; Rodríguez, Mayra C.; Díaz-Godínez, César; Laclette, Juan P.; Becker, Ingeborg; Carrero, Julio C.

    2016-01-01

    Neutrophil defense mechanisms include phagocytosis, degranulation and the formation of extracellular traps (NET). These networks of DNA are triggered by several immune and microbial factors, representing a defense strategy to prevent microbial spread by trapping/killing pathogens. This may be important against Entamoeba histolytica, since its large size hinders its phagocytosis. The aim of this study was to determine whether E. histolytica and their lipopeptidophosphoglycan (EhLPPG) induce the formation of NETs and the outcome of their interaction with the parasite. Our data show that live amoebae and EhLPPG, but not fixed trophozoites, induced NET formation in a time and dose dependent manner, starting at 5 min of co-incubation. Although immunofluorescence studies showed that the NETs contain cathelicidin LL-37 in close proximity to amoebae, the trophozoite growth was only affected when ethylene glycol tetra-acetic acid (EGTA) was present during contact with NETs, suggesting that the activity of enzymes requiring calcium, such as DNases, may be important for amoeba survival. In conclusion, E. histolytica trophozoites and EhLPPG induce in vitro formation of human NETs, which did not affect the parasite growth unless a chelating agent was present. These results suggest that NETs may be an important factor of the innate immune response during infection with E. histolytica. PMID:27415627

  15. Characterization of yeast extracellular vesicles: evidence for the participation of different pathways of cellular traffic in vesicle biogenesis.

    Directory of Open Access Journals (Sweden)

    Débora L Oliveira

    Full Text Available BACKGROUND: Extracellular vesicles in yeast cells are involved in the molecular traffic across the cell wall. In yeast pathogens, these vesicles have been implicated in the transport of proteins, lipids, polysaccharide and pigments to the extracellular space. Cellular pathways required for the biogenesis of yeast extracellular vesicles are largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: We characterized extracellular vesicle production in wild type (WT and mutant strains of the model yeast Saccharomyces cerevisiae using transmission electron microscopy in combination with light scattering analysis, lipid extraction and proteomics. WT cells and mutants with defective expression of Sec4p, a secretory vesicle-associated Rab GTPase essential for Golgi-derived exocytosis, or Snf7p, which is involved in multivesicular body (MVB formation, were analyzed in parallel. Bilayered vesicles with diameters at the 100-300 nm range were found in extracellular fractions from yeast cultures. Proteomic analysis of vesicular fractions from the cells aforementioned and additional mutants with defects in conventional secretion pathways (sec1-1, fusion of Golgi-derived exocytic vesicles with the plasma membrane; bos1-1, vesicle targeting to the Golgi complex or MVB functionality (vps23, late endosomal trafficking revealed a complex and interrelated protein collection. Semi-quantitative analysis of protein abundance revealed that mutations in both MVB- and Golgi-derived pathways affected the composition of yeast extracellular vesicles, but none abrogated vesicle production. Lipid analysis revealed that mutants with defects in Golgi-related components of the secretory pathway had slower vesicle release kinetics, as inferred from intracellular accumulation of sterols and reduced detection of these lipids in vesicle fractions in comparison with WT cells. CONCLUSIONS/SIGNIFICANCE: Our results suggest that both conventional and unconventional pathways of secretion are

  16. Marine derived polysaccharides for biomedical applications: chemical modification approaches.

    Science.gov (United States)

    d'Ayala, Giovanna Gomez; Malinconico, Mario; Laurienzo, Paola

    2008-09-03

    Polysaccharide-based biomaterials are an emerging class in several biomedical fields such as tissue regeneration, particularly for cartilage, drug delivery devices and gelentrapment systems for the immobilization of cells. Important properties of the polysaccharides include controllable biological activity, biodegradability, and their ability to form hydrogels. Most of the polysaccharides used derive from natural sources; particularly, alginate and chitin, two polysaccharides which have an extensive history of use in medicine, pharmacy and basic sciences, and can be easily extracted from marine plants (algae kelp) and crab shells, respectively. The recent rediscovery of poly-saccharidebased materials is also attributable to new synthetic routes for their chemical modification, with the aim of promoting new biological activities and/or to modify the final properties of the biomaterials for specific purposes. These synthetic strategies also involve the combination of polysaccharides with other polymers. A review of the more recent research in the field of chemical modification of alginate, chitin and its derivative chitosan is presented. Moreover, we report as case studies the results of our recent work concerning various different approaches and applications of polysaccharide-based biomaterials, such as the realization of novel composites based on calcium sulphate blended with alginate and with a chemically modified chitosan, the synthesis of novel alginate-poly(ethylene glycol) copolymers and the development of a family of materials based on alginate and acrylic polymers of potential interest as drug delivery systems.

  17. Marine Derived Polysaccharides for Biomedical Applications: Chemical Modification Approaches

    Directory of Open Access Journals (Sweden)

    Paola Laurienzo

    2008-09-01

    Full Text Available Polysaccharide-based biomaterials are an emerging class in several biomedical fields such as tissue regeneration, particularly for cartilage, drug delivery devices and gelentrapment systems for the immobilization of cells. Important properties of the polysaccharides include controllable biological activity, biodegradability, and their ability to form hydrogels. Most of the polysaccharides used derive from natural sources; particularly, alginate and chitin, two polysaccharides which have an extensive history of use in medicine, pharmacy and basic sciences, and can be easily extracted from marine plants (algae kelp and crab shells, respectively. The recent rediscovery of poly-saccharidebased materials is also attributable to new synthetic routes for their chemical modification, with the aim of promoting new biological activities and/or to modify the final properties of the biomaterials for specific purposes. These synthetic strategies also involve the combination of polysaccharides with other polymers. A review of the more recent research in the field of chemical modification of alginate, chitin and its derivative chitosan is presented. Moreover, we report as case studies the results of our recent work concerning various different approaches and applications of polysaccharide-based biomaterials, such as the realization of novel composites based on calcium sulphate blended with alginate and with a chemically modified chitosan, the synthesis of novel alginate-poly(ethylene glycol copolymers and the development of a family of materials based on alginate and acrylic polymers of potential interest as drug delivery systems.

  18. Correlation Between Chain Architecture and Hydration Water Structure in Polysaccharides.

    Science.gov (United States)

    Grossutti, Michael; Dutcher, John R

    2016-03-14

    The physical properties of confined water can differ dramatically from those of bulk water. Hydration water associated with polysaccharides provides a particularly interesting example of confined water, because differences in polysaccharide structure provide different spatially confined environments for water sorption. We have used attenuated total reflection infrared (ATR-IR) spectroscopy to investigate the structure of hydration water in films of three different polysaccharides under controlled relative humidity (RH) conditions. We compare the results obtained for films of highly branched, dendrimer-like phytoglycogen nanoparticles to those obtained for two unbranched polysaccharides, hyaluronic acid (HA), and chitosan. We find similarities between the water structuring in the two linear polysaccharides and significant differences for phytoglycogen. In particular, the results suggest that the high degree of branching in phytoglycogen leads to a much more well-ordered water structure (low density, high connectivity network water), indicating the strong influence of chain architecture on the structuring of water. These measurements provide unique insight into the relationship between the structure and hydration of polysaccharides, which is important for understanding and exploiting these sustainable nanomaterials in a wide range of applications.

  19. Oral versus postingestive origin of polysaccharide appetite in the rat.

    Science.gov (United States)

    Sclafani, A; Nissenbaum, J W

    1987-01-01

    Previous studies have revealed that rats consume substantial amounts of polysaccharide solutions, even if the solutions are made bitter with the addition of sucrose octa acetate (SOA). The present experiment used the gastric sham-feeding preparation to determine if it is the orosensory or postingestive properties of polysaccharides that motivate rats to consume polysaccharide (Polycose) solutions. In Experiment 1, food deprived rats sham fed less of a 0.05% SOA + 32% Polycose solution than they did of a 32% glucose solution, but their SOA-Polycose intake was still considerable (44 ml/hr). The same rats refused to sham feed SOA-gum and SOA-sugar solutions that were similar to the SOA-Polycose solution in bitter taste, viscosity and free sugar content. In Experiment 2, rats sham fed as much of a 32% Polycose solution as they did of a 32% sucrose solution. Despite the gastric fistula, some of the ingested Polycose was absorbed as evidenced by an increase in the rats' blood glucose levels. The addition of acarbose, a drug that inhibits polysaccharide digestion, to the Polycose solution blocked the increase in blood glucose, but did not reduce the rats' sham feeding of the solution. These findings indicate that it is the orosensory (presumably taste) properties of polysaccharide solutions, not their postingestive effects, that initially attract rats to the solutions. The results question the assumption that polysaccharides are "tasteless" to animals.

  20. Health benefits of algal polysaccharides in human nutrition.

    Science.gov (United States)

    Mišurcová, Ladislava; Škrovánková, Soňa; Samek, Dušan; Ambrožová, Jarmila; Machů, Ludmila

    2012-01-01

    The interest in functional food, both freshwater and marine algal products with their possible promotional health effects, increases also in regions where algae are considered as rather exotic food. Increased attention about algae as an abundant source of many nutrients and dietary fiber from the nutrition point of view, as well as from the scientific approaches to explore new nutraceuticals and pharmaceuticals, is based on the presence of many bioactive compounds including polysaccharides extracted from algal matter. Diverse chemical composition of dietary fiber polysaccharides is responsible for their different physicochemical properties, such as their ability to be fermented by the human colonic microbiota resulted in health benefit effects. Fundamental seaweed polysaccharides are presented by alginates, agars, carrageenans, ulvanes, and fucoidans, which are widely used in the food and pharmaceutical industry and also in other branches of industry. Moreover, freshwater algae and seaweed polysaccharides have emerged as an important source of bioactive natural compounds which are responsible for their possible physiological effects. Especially, sulfate polysaccharides exhibit immunomodulatory, antitumor, antithrombotic, anticoagulant, anti-mutagenic, anti-inflammatory, antimicrobial, and antiviral activities including anti-HIV infection, herpes, and hepatitis viruses. Generally, biological activity of sulfate polysaccharides is related to their different composition and mainly to the extent of the sulfation of their molecules. Significant attention has been recently focused on the use of both freshwater algae and seaweed for developing functional food by reason of a great variety of nutrients that are essential for human health.