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MALDI and polysaccharide

Cosima Damiana Calvano, Antonio Monopoli, Tommaso R I Cataldi, Francesco Palmisano
Since its introduction in the 1980s, matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) has gained a prominent role in the analysis of high molecular weight biomolecules such as proteins, peptides, oligonucleotides, and polysaccharides. Its application to low molecular weight compounds has remained for long time challenging due to the spectral interferences produced by conventional organic matrices in the low m/z window. To overcome this problem, specific sample preparation such as analyte/matrix derivatization, addition of dopants, or sophisticated deposition technique especially useful for imaging experiments, have been proposed...
April 23, 2018: Analytical and Bioanalytical Chemistry
Evgeny Vinogradov, Irina Sadovskaya, Pascal Courtin, Saulius Kulakauskas, Thierry Grard, Jennifer Mahony, Douwe van Sinderen, Marie-Pierre Chapot-Chartier
In the lactic acid bacterium Lactococcus lactis, a cell wall polysaccharide (CWPS) is the bacterial receptor of the majority of infecting bacteriophages. The diversity of CWPS structures between strains explains, at least partially, the narrow host range of lactococcal phages. In the present work, we studied the polysaccharide components of the cell wall of the prototype L. lactis subsp. lactis strain IL1403. We identified a rhamnose-rich complex polysaccharide, carrying a glycerophosphate substitution, as the major component...
June 15, 2018: Carbohydrate Research
Sineenart Sengyee, Sung Hwan Yoon, Suporn Paksanont, Thatcha Yimthin, Vanaporn Wuthiekanun, Direk Limmathurotsakul, T Eoin West, Robert K Ernst, Narisara Chantratita
Burkholderia pseudomallei is an environmental bacterium that causes melioidosis, a major community-acquired infection in tropical regions. Melioidosis presents with a range of clinical symptoms, is often characterized by a robust inflammatory response, may relapse after treatment, and results in high mortality rates. Lipopolysaccharide (LPS) of B. pseudomallei is a potent immunostimulatory molecule comprised of lipid A, core, and O-polysaccharide (OPS) components. Four B. pseudomallei LPS types have been described based on SDS-PAGE patterns that represent the difference of OPS-type A, type B, type B2 and rough LPS...
February 2018: PLoS Neglected Tropical Diseases
L García-Calvo, R V Ullán, M Fernández-Aguado, A M García-Lino, R Balaña-Fouce, C Barreiro
Lignocellulosic plant biomass is the most abundant carbon source in the planet, which makes it a potential substrate for biorefinery. It consists of polysaccharides and other molecules with applications in pharmaceutical, food and feed, cosmetics, paper and textile industries. The exploitation of these resources requires the hydrolysis of the plant cell wall, which is a complex process. Aiming to discover novel fungal natural isolates with lignocellulolytic capacities, a screening for feruloyl esterase activity was performed in samples taken from different metal surfaces...
April 15, 2018: Journal of Proteomics
Thais Gonçalves Ferreira, Camilla Nunes Dos Reis Trindade, Petra Bell, André Teixeira-Ferreira, Jonas E Perales, Rossiane C Vommaro, Regina Maria Cavalcanti Pilotto Domingues, Eliane de Oliveira Ferreira
BACKGROUND: Members of the Bacteroides fragilis group are the most important components of the normal human gut microbiome, but are also major opportunistic pathogens that are responsible for significant mortality, especially in the case of bacteraemia and other severe infections, such as intra-abdominal abscesses. Up to now, several virulence factors have been described that might explain the involvement of B. fragilis in these infections. The secretion of extracellular membrane vesicles (EMVs) has been proposed to play a role in pathogenesis and symbiosis in gram-negative bacteria, by releasing soluble proteins and other molecules...
March 2018: Memórias do Instituto Oswaldo Cruz
Khawla Ben Jeddou, Fatma Bouaziz, Claire Boisset Helbert, Oumèma Nouri-Ellouz, Sameh Maktouf, Semia Ellouz-Chaabouni, Raoudha Ellouz-Ghorbel
Enzymatic hydrolysis of water-soluble polysaccharides from potato peel waste (PPPW) generates low molecular weight oligosaccharides with a yield of 63%. The oligosaccharides generated from potato peel polysaccharides (OPPP) were purified by Superdex-30 column. The results showed the presence of 8 peaks (OPPP1-OPPP8). The identification of all the fractions by chromatography analysis (GC-FID) illustrated that the most prominent residues were glucose with traces of galactose, arabinose and rhamnose. Finally, mass spectrometry (MALDI-ToF-ToF) analysis showed that the generated oligosaccharides were heterogeneous and contained different degree of polymerization (DP)...
June 2018: International Journal of Biological Macromolecules
Jun Liu, Ann-Sofie Leppänen, Victor Kisonen, Stefan Willför, Chunlin Xu, Francisco Vilaplana
Accurate determination of the distribution of substitutions in the primary molecular structure of heteropolysaccharides and their derivatives is a prerequisite for their increasing application in the pharmaceutical and biomedical fields, which is unfortunately hindered due to the lack of effective analytical techniques. Acetylated galactoglucomannan (GGM) is an abundant plant polysaccharide as the main hemicellulose in softwoods, and therefore constitutes an important renewable resource from lignocellulosic biomass for the development of bioactive and functional materials...
June 2018: International Journal of Biological Macromolecules
Vladimír Puchart, Lucia Fraňová, Kristian B R Mørkeberg Krogh, Tine Hoff, Peter Biely
Most studies of the mode of action of industrially important endoxylanases have been done on alkali extracted-plant xylan. In just few cases, the native form of the polysaccharide, acetylated xylan, was used as a substrate. In this work action of xylanases belonging to three glycoside hydrolase families, GH10, GH11, and GH30 was investigated on acetylglucuronoxylan directly in hardwood cell walls. Powdered eucalyptus wood was used as xylanase substrate. Enzyme-generated fragments were characterized by TLC, MALDI ToF MS, and NMR spectroscopy...
February 2018: Applied Microbiology and Biotechnology
Shengjie Liu, Jiadong Gao, Zhongjian Chen, Xiaoyan Qiao, Hualin Huang, Baiyuan Cui, Qingfeng Zhu, Zhangyan Dai, Hualing Wu, Yayan Pan, Chengwei Yang, Jun Liu
BACKGROUND: A recently discovered tea [Camellia sinensis (L.) O. Kuntze] cultivar can generate tender shoots in winter. We performed comparative proteomics to analyze the differentially accumulated proteins between winter and spring tender shoots of this clonal cultivar to reveal the physiological basis of its evergrowing character during winter. RESULTS: We extracted proteins from the winter and spring tender shoots (newly formed two leaves and a bud) of the evergrowing tea cultivar "Dongcha11" respectively...
November 20, 2017: BMC Plant Biology
Ai Ling Ho, Ondrej Kosik, Alison Lovegrove, Dimitris Charalampopoulos, Robert A Rastall
Xylo-oligosaccharides and xylo-polysaccharides (XOS, XPS) produced by autohydrolysis of the fibre from oil palm empty fruit bunches (OPEFB) were purified using gel filtration chromatography to separate the XOS and XPS from the crude autohydrolysis liquor. Six mixed fractions of refined XOS and XPS with average degree of polymerisation (avDP) of 4-64 were obtained. These were characterised in terms of their composition and size by HPLC, MALDI-ToF-MS (selected fractions) and carbohydrate gel electrophoresis (PACE)...
January 1, 2018: Carbohydrate Polymers
Lifeng Liu
Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) has become an important tool for the analysis of biomolecules, such as DNA, peptides, and oligosaccharides. This technique has been developed as a rapid, sensitive, and accurate means for analyzing cell wall polysaccharide structures. Here, we describe a method using mass spectrometry to provide xyloglucan composition and structure information of Brachypodium plants which will be useful for functional characterization of xyloglucan biosynthesis pathway in Brachypodium distachyon...
2018: Methods in Molecular Biology
Xiaoyong Zhao, Shanshan Shen, Datong Wu, Pengfei Cai, Yuanjiang Pan
Analysis of carbohydrates based on matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is still challenging and researchers have been devoting themselves to efficient matrices discovery. In the present study, the design, synthesis, qualitative and quantitative performance of non-derivative ionic liquid matrices (ILMs) were reported. DHB/N-methylaniline (N-MA) and DHB/N-ethylaniline (N-EA), performing best for carbohydrate detection, have been screened out. The limit of detection for oligosaccharide provided by DHB/N-MA and DHB/N-EA were as low as 10 fmol...
September 8, 2017: Analytica Chimica Acta
Vijayakumar Poondla, Rajasekhar Chikati, Monika Kallubai, Vidyasagar Chennupati, Rajagopal Subramanyam, Vijaya Sarathi Reddy Obulam
Earlier, low-temperature-active polygalacturonase isoforms from Saccharomyces cerevisiae PVK4 were isolated and purified. Substrate specificity of polygalacturonase isoforms indicated high affinity for pectins and very low enzyme activity towards non-pectic polysaccharides. To characterize the polygalacturonase isoforms, biochemical, spectral, and in silico approaches were used. The apparent Km and Vmax values for hydrolysis of pectin and galacturonic acid were 0.31 mg/ml and 3.15 mmol min/mg, respectively...
October 2017: 3 Biotech
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
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