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MALDI and polysaccharide

Xiaoyong Zhao, Jinying Li, Yaqin Liu, Datong Wu, Pengfei Cai, Yuanjiang Pan
As a folk medicine, Botrychium ternatum has been used for thousands of years in China. In the present work, a water soluble polysaccharide BTp1 was extracted and purified from B. ternatum. Based on the MALDI matrix 3-aminoquinoline-α-cyano-4-hydroxycinnamic acid, the molecular weight of BTp1 was determined to be 11638Da directly. Monosaccharide analysis showed that BTp1 was composed of arabinose (Ara). Combining enzymatic hydrolysis and subsequent MALDI-TOF analysis, a linear backbone of BTp1, consisted of (1→5)-linked α-l-Araf, was inferred quickly...
September 1, 2017: Carbohydrate Polymers
Majela González-Miro, Laura Rodríguez-Noda, Mildrey Fariñas-Medina, Dagmar García-Rivera, Vicente Vérez-Bencomo, Bernd H A Rehm
Streptococcus pneumoniae is a human pathogen responsible for the majority of childhood pneumonia and media otitis cases worldwide. The diversity of its capsular polysaccharides (CPS) results in more than 91 serotypes of which at least 23 are virulent. Various CPS conjugated to immunogenic carrier proteins are currently licensed and provide protection against the infection caused by the respective serotypes but not against new and emerging virulent serotypes. In this study, we considered the conserved protein antigen PsaA, the pneumococcal surface adhesin A, in order to overcome the limitations of CPS antigens...
April 2017: Heliyon
Yuqiang Zheng, Xuemei Zhang, Xiaofang Wang, Libin Wang, Jinghui Zhang, Yibing Yin
The capsular polysaccharide (CPS) of Streptococcus pneumoniae is the main virulence factors required for effective colonization and invasive disease. The capacity to regulate CPS production at the transcriptional level is critical for the survival of S. pneumoniae in different host niches, but little is known about the transcription regulators of cps locus. In the present study, we isolated and identified the response regulator ComE, the master competence switch in transformation of S. pneumoniae, as a transcriptional regulator of cps locus by DNA affinity chromatography-pulldown, MALDI-TOF mass spectrometry (MS) and electrophoretic mobility shift assay (EMSA)...
2017: Frontiers in Microbiology
Umut Şahar, Remziye Deveci
Sea urchin eggs are surrounded by a carbohydrate-rich layer, termed the jelly coat, that consists of polysaccharides and glycoproteins. In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS)...
May 2017: Molecular Reproduction and Development
Jolanda M van Munster, Baptiste Thomas, Michel Riese, Adrienne L Davis, Christopher J Gray, David B Archer, Sabine L Flitsch
Renewables-based biotechnology depends on enzymes to degrade plant lignocellulose to simple sugars that are converted to fuels or high-value products. Identification and characterization of such lignocellulose degradative enzymes could be fast-tracked by availability of an enzyme activity measurement method that is fast, label-free, uses minimal resources and allows direct identification of generated products. We developed such a method by applying carbohydrate arrays coupled with MALDI-ToF mass spectrometry to identify reaction products of carbohydrate active enzymes (CAZymes) of the filamentous fungus Aspergillus niger...
February 21, 2017: Scientific Reports
Leonard Barnabas, N M R Ashwin, Kalimuthu Kaverinathan, Anna Rita Trentin, Micaela Pivato, Amalraj Ramesh Sundar, Palaniyandi Malathi, Rasappa Viswanathan, Paolo Carletti, Giorgio Arrigoni, Antonio Masi, Ganesh Kumar Agrawal, Randeep Rakwal
Sporisorium scitamineum, the sugarcane smut pathogen, relies predominantly on its secretome to successfully colonise its host, in accordance with other related smut fungi. Considering the significance of deciphering its secretome, we have examined alterations in the in vitro secretome of S. scitamineum in response to synthetic and sugarcane meristem tissue-amended growth media, so as to identify host signal responsive secretory proteins. Secretory proteins that were differentially abundant and exclusively secreted in response to host extract media were identified by two-dimensional gel electrophoresis coupled with MALDI-TOF/TOF MS...
March 2017: Fungal Biology
Clara Granzotto, Ken Sutherland
This paper reports an improved method for the identification of Acacia gum in cultural heritage samples using matrix assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) after enzymatic digestion of the polysaccharide component. The analytical strategy was optimized using a reference Acacia gum (gum arabic, sp. A. senegal) and provided an unambiguous MS profile of the gum, characterized by specific and recognized oligosaccharides, from as little as 0.1 μg of material. The enhanced experimental approach with reduced detection limit was successfully applied to the analysis of naturally aged (∼80 year) gum arabic samples, pure and mixed with lead white pigment, and allowed the detection of gum arabic in samples from a late painting (1949/1954) by Georges Braque in the collection of the Art Institute of Chicago...
March 7, 2017: Analytical Chemistry
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...
February 1, 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...
May 19, 2017: 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
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