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MALDI and polysaccharide

Iwona Komaniecka, Adam Choma, Katarzyna Zamlynska, Anna Sroka-Bartnicka, Pawel Sowinski
Oligotropha carboxidovorans strain OM5 (previously known as Pseudomonas carboxydovorans OM5) is a rod-shaped Gram-negative bacterium isolated from wastewater. This bacterium is able to live in aerobic and, facultatively, in autotrophic conditions. For autotrophic growth, the bacteria can utilize carbon monoxide or hydrogen as a source of energy. The O-specific polysaccharide isolated from O. carboxidovorans OM5 lipopolysaccharide was structurally characterized using chemical analyses, 1D and 2D NMR spectroscopy, and MALDI-TOF mass spectrometry techniques...
January 3, 2017: Carbohydrate Research
Xudong Sun, Jing Dong, Jinan Li, Mingliang Ye, Weibing Zhang, Junjie Ou
A facile approach for preparation of hydrophilic polysaccharide-functionalized macroporous adsorption resin (MAR) was presented. Polydisperse MAR with approximately 250μm in diameter was synthesized via suspension polymerization and then modified with polysaccharides, sodium hyaluronate (HA) and chitosan (CS), through layer-by-layer (LbL) self-assembly process. The preparation method was facile, low cost and easy to expand production from tens of grams to one kilogram. The resulting MAR@(HA/CS)20 possesses highly hydrophilic property and rapid adsorption behavior...
December 16, 2016: Journal of Chromatography. A
Stanislav D Anastyuk, Natalia M Shevchenko, Roza V Usoltseva Menshova, Artyom S Silchenko, Pavel A Zadorozhny, Pavel S Dmitrenok, Svetlana P Ermakova
A fucoidan ScF from brown alga Saccharina cichorioides was extracted, purified and partially depolymerized by autohydrolysis at 37°C for 24, 48 and 72h. Supernatant (SN) and pellet (PL) fractions were obtained by ethanol precipitation of each sample. Unlike spectral data of ScF, NMR of PL derivatives clearly suggested the structure: 1,3-linked α-l-Fucp-4-OSO3(-) repeating unit. Molecular weights (MWs) of PL fractions were 30, 26 and 18kDa for 24, 48 and 72h of autohydrolyis, respectively. MALDI-TOFMS, size-exclusion HPLC and carbohydrate polyacrylamide-gel electrophoresis (C-PAGE) indicated a similarity of SN mixtures...
February 10, 2017: Carbohydrate Polymers
Kadda Hachem, Céline Faugeron, Meriem Kaid-Harche, Vincent Gloaguen
Xylan-type polysaccharides were isolated from the leaves of Argania spinosa (L.) Skeels collected in the Tindouf area (southwestern Algeria). Xylan fractions were obtained by sequential alkaline extractions and purified on Sepharose CL-4B. The xylan structure was investigated by enzymatic hydrolysis with an endo-β(1→4)-xylanase followed by chromatography of the resulting fragments on Biogel P2, characterization by sugar analysis and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS )...
November 21, 2016: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
P Domizio, Y Liu, L F Bisson, D Barile
The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. A higher content of polysaccharide was found in media fermented by Schizosaccharomyces japonicus with respect to that of Schizosaccharomyces pombe...
February 2017: Food Microbiology
Rodell C Barrientos, Melody M Clerigo, Anamy Ma C Paano
We report for the first time the extraction, isolation, and the proposed structure of a polysaccharide from the fruiting bodies of Daedalea quercina. The monosaccharide composition of D. quercina isolate (DQW1Pa1) was mainly glucose as identified using GC-MS. FTIR-ATR spectroscopy and absolute configuration studies showed that this polysaccharide is a β-d-glucan. Its average molecular weight obtained using size exclusion chromatography was 1.6×10(4)Da, consistent with glucans derived from the order Polyporaceae...
December 2016: International Journal of Biological Macromolecules
Jun Liu, Victor Kisonen, Stefan Willför, Chunlin Xu, Francisco Vilaplana
Plant polysaccharides constitute arguably the most complex family of biomacromolecules in terms of the stereochemistry and regiochemistry of their intramolecular linkages. The chemical modification of such polysaccharides introduces an additional level of complexity for structural determinations. We have developed an integrated analytical procedure combining selective enzymatic hydrolysis, hydrophilic interaction liquid chromatography (HILIC), and mass spectrometry (MS) to describe the substitution pattern of xyloglucan (XyG) and its chemo-enzymatic derivatives (cationic, anionic, and benzyl aminated)...
September 9, 2016: Journal of Chromatography. A
Ariane Nimptsch, Jürgen Schiller
RATIONALE: If carbohydrates are investigated by nuclear magnetic resonance (NMR) spectroscopy, they are normally dissolved in deuterated solvents, such as D2 O. The incorporation of deuterium leads to a high complexity of subsequently recorded mass spectra and reduced sensitivity because different deuterated ions become detectable. Here, we demonstrate that the applied matrix-assisted laser desorption/ionization (MALDI) matrix solution has a considerable impact on the observed isotopic distribution...
October 15, 2016: Rapid Communications in Mass Spectrometry: RCM
Huihuang H Ding, Steve W Cui, H Douglas Goff, Jie Chen, Qingbin Guo, Qi Wang
The structure of ethanol precipitated fraction from 1M KOH extracted flaxseed kernel polysaccharides (KPI-EPF) was studied for better understanding the molecular structures of flaxseed kernel cell wall polysaccharides. Based on methylation/GC-MS, NMR spectroscopy, and MALDI-TOF-MS analysis, the dominate sugar residues of KPI-EPF fraction comprised of (1,4,6)-linked-β-d-glucopyranose (24.1mol%), terminal α-d-xylopyranose (16.2mol%), (1,2)-α-d-linked-xylopyranose (10.7mol%), (1,4)-β-d-linked-glucopyranose (10...
October 20, 2016: Carbohydrate Polymers
Tim Steffens, Frank-Jörg Vorhölter, Marco Giampà, Gerd Hublik, Alfred Pühler, Karsten Niehaus
BACKGROUND: The exopolysaccharide xanthan is a natural product which is extensively used in industry. It is a thickening agent in many fields, from oil recovery to the food sector. Xanthan is produced by the Gram negative bacterium Xanthomonas campestris pv. campestris (Xcc). We analyzed the lipopolysaccharide (LPS) of three mutant strains of the Xcc wild type B100 to distinguish if the xanthan production can be increased when LPS biosynthesis is affected. RESULTS: The Xcc B100 O-antigen (OA) is composed of a linear main chain of rhamnose residues with N-acetylfucosamine (FucNAc) side branches at every second rhamnose...
May 23, 2016: BMC Microbiology
Li Yu, Shuqin Xu, Chao Deng, Hui Li, Qingsheng Yang, Zhenghong Xu, Jinghua Chen
In this study, an acidic hetero-exopolysaccharide, designated as BMPS, was isolated from the culture supernatant of bacteria Bacillus mucilaginosus SM-01 by ion-exchange and size-exclusion chromatography. The molecular weight of BMPS was measured to be 2.67×10(6)Da. To analyze the structure of the polysaccharide, the low-molecular weight fragment (BMPS-H) of BMPS was obtained under mild hydrolysis. BMPS-H was determined to be a liner neutral fraction with an average molecular weight of 1621Da. Monosaccharide analysis indicated that BMPS-H contained glucose and mannose in a molar ratio of 1...
August 1, 2016: Carbohydrate Polymers
Sunil Ratnayake, Kristina Ford, Antony Bacic
This protocol describes the specific techniques used for the characterization of reducing end (RE) and internal region glycosyl sequence(s) of heteroxylans. De-starched wheat endosperm cell walls were isolated as an alcohol-insoluble residue (AIR)(1) and sequentially extracted with water (W-sol Fr) and 1 M KOH containing 1% NaBH4 (KOH-sol Fr) as described by Ratnayake et al. (2014)(2). Two different approaches (see summary in Figure 1) are adopted. In the first, intact W-sol AXs are treated with 2AB to tag the original RE backbone chain sugar residue and then treated with an endoxylanase to generate a mixture of 2AB-labelled RE and internal region reducing oligosaccharides, respectively...
March 24, 2016: Journal of Visualized Experiments: JoVE
Lennart Leßmeier, Rabeaa S Alkhateeb, Fabian Schulte, Tim Steffens, Tobias Pascal Loka, Alfred Pühler, Karsten Niehaus, Frank-Jörg Vorhölter
At a molecular level, the regulation of many important cellular processes is still obscure in xanthomonads, a bacterial group of outstanding relevance as world-wide plant pathogens and important for biotechnology as producers of the polysaccharide xanthan. Transcriptome analysis indicated a sucrose-dependent regulation of 18 genes in Xanthomonas campestris pv. campestris (Xcc) B100. The expression of 12 of these genes was clearly increased in the presence of sucrose. Only part of these genes was obviously involved in sucrose utilization...
August 20, 2016: Journal of Biotechnology
Yangyang Chai, Guibin Wang, Lili Fan, Min Zhao
The anti-tumor properties of fungal polysaccharides have gained significant recognition in Asia and tropical America. In this study, the differential expression of proteins in normal HepG2 cells and those treated with polysaccharides that had been isolated from Phellinus linteus (PL), Ganoderma lucidum (GL) and Auricularia auricula (AA) was investigated. Using two-dimensional electrophoresis (2DE), a total of 104 protein spots were determined to be overexpressed in these cells compared with noncancerous regions...
2016: Scientific Reports
Abida Sultan, Birgit Andersen, Birte Svensson, Christine Finnie
Cereal grains are colonized by a microbial community that actively interacts with the plant via secretion of various enzymes, hormones, and metabolites. Microorganisms decompose plant tissues by a collection of depolymerizing enzymes, including β-1,4-xylanases, that are in turn inhibited by plant xylanase inhibitors. To gain insight into the importance of the microbial consortia and their interaction with barley grains, we used a combined gel-based (2-DE coupled to MALDI-TOF-TOF MS) and gel-free (LC-MS/MS) proteomics approach complemented with enzyme activity assays to profile the surface-associated proteins and xylanolytic activities of two barley cultivars...
April 1, 2016: Journal of Proteome Research
Tzu-Wen Liang, Shih-Chun Tseng, San-Lang Wang
Natural polysaccharides have received much attention due to their wide range of applications. Although most microbial exopolysaccharides (EPSs) use sugars as the major carbon source, such as glucose or sucrose, in this study, EPSs were induced from a squid pen powder (SPP)-containing medium by Paenibacillus mucilaginosus TKU032, a bacterial strain isolated from Taiwanese soil. Under the optimal culture conditions, the maximum EPS yield (14.8 g/L) was obtained. MALDI-TOF MS analysis of an EPS fraction purified by gel filtration revealed two mass peaks with molecular weights of ∼1...
February 19, 2016: Marine Drugs
Marta López-García, María Sonia Dopico García, José Manuel López Vilariño, María Victoria González Rodríguez
In this work MALDI-TOF mass spectroscopy was investigated to characterise the β-glucan profiles of several commercial health supplements, without any derivatisation or purification pre-treatment. The effect of two solvents (water and dimethyl sulfoxide) and two MALDI matrices (2,5-dihydroxybenzoic acid and 2',4',6'-trihydroxyacetophenone) was first evaluated on dextran standards. MALDI-TOF was found as a useful and quick technique to obtain structural information of diverse food supplements based on mushroom extracts...
May 15, 2016: Food Chemistry
Xiaomeng Dai, Libo Yang, Zhiyong Zheng, Haiqin Chen, Xiaobei Zhan
OBJECTIVE: Curdlan is produced by Agrobacterium sp. ATCC 31749 under nitrogen limiting condition. The biosynthesis of crudlan is a typical aerobic bioprocess, and the production of curdlan would be severely restricted under micro-aerobic and anoxic conditions. Proteomic analysis of Agrobacterium sp. was conducted to investigate the effect of dissolved oxygen on the crucial enzymes involved in curdlan biosynthesis. METHODS: Two-dimensional gel electrophoresis was performed to separate and visualize the differential expression of the intracellular proteins extracted from Agrobacterium sp...
August 4, 2015: Wei Sheng Wu Xue Bao, Acta Microbiologica Sinica
Fábio Faria-Oliveira, Joana Carvalho, Célia Ferreira, Maria Luisa Hernáez, Concha Gil, Cândida Lucas
BACKGROUND: Saccharomyces cerevisiae multicellular communities are sustained by a scaffolding extracellular matrix, which provides spatial organization, and nutrient and water availability, and ensures group survival. According to this tissue-like biology, the yeast extracellular matrix (yECM) is analogous to the higher Eukaryotes counterpart for its polysaccharide and proteinaceous nature. Few works focused on yeast biofilms, identifying the flocculin Flo11 and several members of the HSP70 in the extracellular space...
2015: BMC Microbiology
Silvia Minoia, Adnane Boualem, Fabien Marcel, Christelle Troadec, Bernard Quemener, Francesco Cellini, Angelo Petrozza, Jacqueline Vigouroux, Marc Lahaye, Filomena Carriero, Abdelhafid Bendahmane
Fruit ripening and softening are key traits for many fleshy fruit. Since cell walls play a key role in the softening process, expansins have been investigated to control fruit over ripening and deterioration. In tomato, expression of Expansin 1 gene, SlExp1, during fruit ripening was associated with fruit softening. To engineer tomato plants with long shelf life, we screened for mutant plants impaired in SlExp1 function. Characterization of two induced mutations, Slexp1-6_W211S, and Slexp1-7_Q213Stop, showed that SlExp1 loss of function leads to enhanced fruit firmness and delayed fruit ripening...
January 2016: Plant Science: An International Journal of Experimental Plant Biology
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