The structure of the extracellular teichoic acids from the allergy-protective bacterium Lactococcus lactis G121

2012 ◽  
Vol 393 (8) ◽  
pp. 749-755
Author(s):  
Kathleen Fischer ◽  
Evgueny Vinogradov ◽  
Buko Lindner ◽  
Holger Heine ◽  
Otto Holst
Keyword(s):  
Lactococcus Lactis ◽  
Molecular Mechanisms ◽  
Cell Envelope ◽  
Teichoic Acid ◽  
Size Exclusion ◽  
Resonance Spectroscopy ◽  
Protective Properties ◽  
Isolation And Characterization ◽  
Exclusion Chromatography

Abstract The Gram-positive bacterium Lactococcus lactis G121 is a farm isolate that protects mice from ovalbumin-induced asthma. To understand the molecular mechanisms of such allergy-protective properties, the isolation and characterization of cell envelope constituents is crucial. Here, structural analyses of the extracellular teichoic acid (EC TA) from L. lactis G121 are presented. Extraction with 0.9% saline afforded a crude TA fraction. Consecutive size exclusion chromatography on Biogel P60 and P10 matrix was performed to purify the sample. Chemical component analyses, high-resolution electrospray ionization Fourier-transformed ion cyclotron mass spectrometry, and nuclear magnetic resonance spectroscopy were conducted for structural elucidation. The EC TA was a poly(glycosylglycerol phosphate) molecule with a repeating unit of -6)-[β-d-Glcp-(1→3)-][α-d-GlcpNAc-(1→4)-]α-d-GalpNAc-(1→3)-β-d-GlcpNAc-(1→2)-glycerol-(1-P-).

scholarly journals Synthesis and characterization of amphiphilic graft copolymers of poly(1,3 dioxolane) macromonomers with styrene and methyl methacrylate

e-Polymers ◽  
2009 ◽  
Vol 9 (1) ◽  
Author(s):  
Hayet Bendaikha ◽  
Gérald Clisson ◽  
Abdelouahad Khoukh ◽  
Jeanne François ◽  
Seghier Ould Kada
Keyword(s):  
Methyl Methacrylate ◽  
Size Exclusion ◽  
Resonance Spectroscopy ◽  
Molecular Weights ◽  
Molar Ratios ◽  
H Nmr ◽  
Amphiphilic Graft Copolymers ◽  
Exclusion Chromatography ◽  
Cationic Ring Opening Polymerization

AbstractMethacrylate-terminated Poly(1,3 dioxolane) (PDXL) macromonomers were synthesized by cationic ring-opening polymerization in the presence of 2- hydroxypropyl methacrylate (2-HPMA) as transfer agent. Molecular weights, polydispersity index and functionality of the PDXL macromonomers were evaluated by size exclusion chromatography (SEC) and 1H nuclear magnetic resonance spectroscopy (1H-NMR). Copolymerizations of PDXL macromonomers, of different molecular weights, with styrene (St) and methyl methacrylate (MMA) were carried out using various feed molar ratios. The resulting polymers confirmed the grafting of PDXL with PS and PMMA by SEC and 1H-NMR Monomer reactivity ratios between the macromonomers and the comonomers were estimated from the copolymerization results. Macromonomer reactivity depends on the comonomer considered. Glass transition temperatures of the copolymers were found to decrease with an increase in the amount of PDXL in the copolymers. The values of Tg depend on the composition and the size of the PDXL grafts.

scholarly journals Methods for Separation and Characterization of Extracellular Vesicles: Results of a Worldwide Survey Performed by the ISEV Rigor and Standardization Subcommittee

Cells ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1955 ◽  
Author(s):  
Felix Royo ◽  
Clotilde Théry ◽  
Juan M. Falcón-Pérez ◽  
Rienk Nieuwland ◽  
Kenneth W. Witwer
Keyword(s):  
Extracellular Vesicles ◽  
Initial Response ◽  
Size Exclusion ◽  
Disease Biomarkers ◽  
Quality Controls ◽  
Isolation And Characterization ◽  
Measurement Results ◽  
Exclusion Chromatography ◽  
Important Objective

Research on extracellular vesicles (EVs) is growing exponentially due to an increasing appreciation of EVs as disease biomarkers and therapeutics, an expanding number of EV-containing materials under study, and application of new preparation, detection, and cargo analysis methods. Diversity of both sources and methodologies imposes challenges on the comparison of measurement results between studies and laboratories. While reference guidelines and minimal requirements for EV research have achieved the important objective of assembling community consensus, it is also essential to understand which methodologies and quality controls are currently being applied, and how usage trends are evolving. As an initial response to this need, the International Society for Extracellular Vesicles (ISEV) performed a worldwide survey in 2015 on “Techniques used for the isolation and characterization of extracellular vesicles” and published the results from this survey in 2016. In 2019, a new survey was performed to assess the changing state of the field. The questionnaire received more than 600 full or partial responses, and the present manuscript summarizes the results of this second worldwide survey. The results emphasize that separation methods such as ultracentrifugation and density gradients are still the most commonly used methods, the use of size exclusion chromatography has increased, and techniques based on tangential flow and microfluidics are now being used by more than 10% of respondents. The survey also reveals that most EV researchers still do not perform sample quality controls before or after isolation of EVs. Finally, the majority of EV researchers emphasize that separation and characterization of EVs should receive more attention.

scholarly journals Isolation and characterization of BanLec-I, a mannoside-binding lectin from Musa paradisiac (banana)

1990 ◽  
Vol 272 (3) ◽  
pp. 721-726 ◽  
Author(s):  
V L Koshte ◽  
W van Dijk ◽  
M E van der Stelt ◽  
R C Aalberse
Keyword(s):  
Cell Proliferation ◽  
Affinity Chromatography ◽  
Molecular Mass ◽  
Size Exclusion ◽  
T Cell Proliferation ◽  
Isolation And Characterization ◽  
Sds Page ◽  
Exclusion Chromatography ◽  
Binding Lectin

A lectin (BanLec-I) from banana (Musa paradisiac) with a binding specificity for oligomannosidic glycans of size classes higher than (Man)6GlcNAc was isolated and purified by affinity chromatography on a Sephadex G-75 column. It did not agglutinate untreated human or sheep erythrocytes, but it did agglutinate rabbit erythrocytes. BanLec-I stimulated T-cell proliferation. On size-exclusion chromatography, BanLec-I has a molecular mass of approx. 27 kDa, and on SDS/PAGE the molecular mass is approx. 13 kDa. The isoelectric point is 7.2-7.5. BanLec-I was found to be very effective as a probe in detecting glycoproteins, e.g. on nitrocellulose blots.

scholarly journals Biochemical and Biophysical Characterization of the dsDNA packaging motor from the Lactococcus lactis bacteriophage asccphi28

2020 ◽  
Author(s):  
Emilio Reyes-Aldrete ◽  
Erik A. Dill ◽  
Cecile Bussetta ◽  
Michal R. Szymanski ◽  
Geoffrey Diemer ◽  
...  
Keyword(s):  
Lactococcus Lactis ◽  
Analytical Ultracentrifugation ◽  
Atp Hydrolysis ◽  
Size Exclusion ◽  
Radius Of Gyration ◽  
Biophysical Characterization ◽  
X Ray ◽  
Symmetric Complex ◽  
Exclusion Chromatography

AbstractDouble-stranded DNA viruses package their genomes into pre-assembled protein procapsids. This process is driven by macromolecular motors that transiently assemble at a unique vertex of the procapsid and utilize homomeric ring ATPases to couple genome encapsidation to ATP hydrolysis. Here we describe biochemical and biophysical characterization of the packaging ATPase from Lactococcus lactis phage asccφ28. Size-exclusion chromatography, analytical ultracentrifugation, small angle x-ray scattering, and negative stain TEM indicate that the ~45 kDa protein formed a 443 kDa cylindrical assembly with a maximum dimension of ~155 Å and radius of gyration of ~54 Å. Together with the dimensions of the crystallographic asymmetric unit from preliminary X-ray diffraction experiments, these results indicate that gp11 forms a decameric D5-symmetric complex consisting of two pentameric rings related by 2-fold symmetry. Additional kinetic analysis shows that recombinantly expressed gp11 has ATPase activity comparable to that of functional ATPase rings assembled on procapsids in other genome packaging systems. Hence, gp11 forms rings in solution that likely reflect the fully assembled ATPases in active virus-bound motor complexes. Whereas ATPase functionality in other dsDNA phage packaging systems requires assembly on viral capsids, the ability to form functional rings in solution imparts gp11 with significant advantages for high resolution structural studies and rigorous biophysical/biochemical analysis.

Isolation and characterization of lignins from Eucalyptus grandis Hill ex Maiden and Eucalyptus globulus Labill. by enzymatic mild acidolysis (EMAL)

Holzforschung ◽  
2008 ◽  
Vol 62 (1) ◽  
pp. 24-30 ◽  
Author(s):  
Anderson Guerra ◽  
Lucian A. Lucia ◽  
Dimitris S. Argyropoulos
Keyword(s):  
Eucalyptus Globulus ◽  
Eucalyptus Grandis ◽  
Raw Material ◽  
Size Exclusion ◽  
Hydroxyl Groups ◽  
Aryl Ether ◽  
Isolation And Characterization ◽  
White Fir ◽  
Exclusion Chromatography

Abstract Despite the growing importance of Eucalyptus wood as raw material for pulp and paper, there is a lack of knowledge on the chemistry of their macromolecular components. The present paper addresses this issue by applying the recently developed protocol for isolating enzymatic mild acidolysis lignins (EMAL) from Eucalyptus grandis, Eucalyptus globulus and the softwood species Douglas fir and white fir, which were used for comparative purposes. The structures of EMALs were investigated by quantitative 31P NMR, DFRC/31P NMR (derivatization followed by reductive cleavage followed by quantitative 31P NMR) and size exclusion chromatography (SEC). Overall, the yields of EMALs isolated from Eucalyptus were higher than those from the softwoods examined. Lignin from E. globulus was found to contain higher contents of arylglycerol-β-aryl ether structures, free phenolic hydroxyl groups and syringyl-type units than lignin from E. grandis. New insights provided by the DFRC/31P NMR revealed that up to 62.2% of arylglycerol-β-aryl ether structures in E. globulus are uncondensed, while in E. grandis the amount of such uncondensed structures was found to be lower than 48%. SEC analyses showed that lignins from E. grandis and softwoods associate in greater extension than lignin from E. globulus.

Structure of a shear-thickening polysaccharide extracted from the New Zealand black tree fern, Cyathea medullaris

2020 ◽  
Author(s):  
M Wee ◽  
M Mastrangelo ◽  
Susan Carnachan ◽  
Ian Sims ◽  
K Goh
Keyword(s):  
New Zealand ◽  
Uronic Acid ◽  
Average Molecular Weight ◽  
Shear Thickening ◽  
Water Soluble ◽  
Size Exclusion ◽  
Resonance Spectroscopy ◽  
Tree Fern ◽  
13C Nuclear Magnetic Resonance ◽  
Exclusion Chromatography

A shear-thickening water-soluble polysaccharide was purified from mucilage extracted from the fronds of the New Zealand black tree fern (Cyathea medullaris or 'mamaku' in Māori) and its structure characterised. Constituent sugar analysis by three complementary methods, combined with linkage analysis (of carboxyl reduced samples) and 1H and 13C nuclear magnetic resonance spectroscopy (NMR) revealed a glucuronomannan comprising a backbone of 4-linked methylesterified glucopyranosyl uronic acid and 2-linked mannopyranosyl residues, branched at O-3 of 45% and at both O-3 and O-4 of 53% of the mannopyranosyl residues with side chains likely comprising terminal xylopyranosyl, terminal galactopyranosyl, non-methylesterified terminal glucopyranosyl uronic acid and 3-linked glucopyranosyl uronic acid residues. The weight-average molecular weight of the purified polysaccharide was ~1.9×106Da as determined by size-exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS). The distinctive rheological properties of this polysaccharide are discussed in relation to its structure. © 2014 Elsevier B.V.

Behavior of xylans from the Eucalyptus species. Part 2. Characterization of 4-O-methylglucuronoxylans isolated from black liquors of kraft pulping of Eucalyptus grandis and E. urophylla

Holzforschung ◽  
2013 ◽  
Vol 67 (2) ◽  
pp. 123-128
Author(s):  
Andréia S. Magaton ◽  
Teresa Cristina F. Silva ◽  
Jorge Luiz Colodette ◽  
Dorila Piló-Veloso ◽  
Flaviana Reis Milagres ◽  
...  
Keyword(s):  
Average Molecular Weight ◽  
Eucalyptus Grandis ◽  
Kraft Pulping ◽  
Raw Material ◽  
Size Exclusion ◽  
Eucalyptus Species ◽  
Exclusion Chromatography ◽  
Ft Ir ◽  
Black Liquors

Abstract 4-O-methylglucuronoxylans isolated from Eucalyptus grandis and Eucalyptus urophylla kraft black liquors (KBLs) were chemically characterized by Fourier transform infrared spectroscopy (FT-IR), size exclusion chromatography (SEC), and nuclear magnetic resonance (NMR) spectroscopy. Doses of alkali charge, expressed as active alkali (AA), were 16, 17, and 18% while the sulfidity was kept at 25%. Kappa numbers of 19.1, 17.5, and 16.1 for E. grandis and 20.4, 16.8, and 15.4 for E. urophylla were obtained. At higher alkali charges, the recovery of xylans from the KBLs was lower and the degree of substitution of xylans with uronic acids decreased. The average molecular weight (Mw) of the recovered xylans was greater under conditions of mild pulping, i.e., in the case of pulps with higher kappa numbers. Mw of xylans ranged from 16.1 to 19.1 kDa for E. grandis and from 15.4 to 20.4 kDa for E. urophylla. The xylans from KBL may be useful as pulp modifying agents or as a raw material for advanced applications.

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