scholarly journals Identification and characterization of glycosyltransferases involved in the synthesis of the side chains of the cell wall pectic polysaccharide rhamnogalacturonan II

2015 ◽  
Author(s):  
Malcolm O'Neill
Keyword(s):  
Cell Wall ◽  
Side Chains ◽  
Pectic Polysaccharide ◽  
Rhamnogalacturonan Ii ◽  
Identification And Characterization

scholarly journals Protocols for Isolating and Characterizing Polysaccharides From 1 Plant Cell Walls: A Case Study Using Rhamnogalacturonan-II

2021 ◽  
Author(s):  
Breeanna Urbanowicz ◽  
William Barnes ◽  
Sabina Koj ◽  
Ian Black ◽  
Stephanie Archer-Hartmann ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Size Exclusion ◽  
Proton Nuclear Magnetic Resonance ◽  
Wall Material ◽  
Resonance Spectroscopy ◽  
Cell Wall Polysaccharide ◽  
Pectic Polysaccharide ◽  
Rhamnogalacturonan Ii

Abstract Background: In plants, there is a large diversity of polysaccharides that comprise the cell wall. Each major type of plant cell wall polysaccharide, including cellulose, hemicellulose, and pectin, has distinct structures and functions that contribute to wall mechanics and influence plant morphogenesis. In recent years, pectin modification and valorization has attracted much attention due to its expanding roles of pectin in biomass deconstruction, food science, material science, and environmental remediation. However, pectin utilization has been limited by our incomplete knowledge of pectin structure. Herein, we present a workflow of principles relevant for the characterization of polysaccharide primary structure using nature’s most complex polysaccharide, rhamnogalacturonan-II (RG-II), as a model.Results: We outline how to isolate RG-II from celery and duckweed cell wall material and red wine using chemical or enzymatic treatments coupled with size-exclusion chromatography. From there, we demonstrate the use of mass spectrometry (MS)-based techniques to determine the glycosyl residue and linkage compositions of the intact RG II molecule and RG-II-derived oligosaccharides including special considerations for labile monosaccharides. In doing so, we demonstrated that in the duckweed Wolffiella repanda the arabinopyranosyl (Arap) residue of side chain B is substituted at O-2 with rhamnose. As RG-II is further modified by non-glycosyl modifications including methyl-ethers, methyl-esters, and acetyl-esters, we then describe ways to use electrospray-MS to identify these moieties on RG-II-derived oligosaccharides. We then explored the utility of proton nuclear magnetic resonance spectroscopy (1H-NMR) in identifying RG-II-specific sugars and non-glycosyl modifications to complement and extend MS-based approaches. Finally, we describe how to assess the factors that affect RG-35 II dimerization using liquid chromatographic and NMR spectroscopic approaches.Conclusions: The complexity of pectic polysaccharide structures has hampered efforts aimed at their valorization. In this work, we used RG-II as a model to demonstrate the steps necessary to isolate and characterize polysaccharides using chromatographic, MS, and NMR techniques. The principles can be applied to the characterization of other saccharide structures and will help inform researchers on how saccharide structure relates to functional properties in the future.

Identification and characterization of a major building block in the cell wall of Saccharomyces cerevisiae

1997 ◽  
Vol 25 (3) ◽  
pp. 856-860 ◽  
Author(s):  
F. M. Klis ◽  
L. H. P. Caro ◽  
J. H. Vossen ◽  
J. C. Kapteyn ◽  
A. F. J. Ram ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Wall ◽  
Building Block ◽  
Identification And Characterization

scholarly journals Pectin Rhamnogalacturonan II: On the “Small Stem with Four Branches” in the Primary Cell Walls of Plants

2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
Beda M. Yapo
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Vascular Plants ◽  
Minor Component ◽  
Primary Cell ◽  
Side Chains ◽  
Complex Cell ◽  
Rhamnogalacturonan Ii ◽  
Branching Point ◽  
A Chain

Rhamnogalacturonan II (RG-II) is a type of block copolymer of complex pectins that represents a quantitatively minor component of the primary cell walls of land (vascular) plants. The structural composition of RG-II is almost totally sequenced and appears to be remarkably conserved in all tracheophytes so far examined. The backbone of RG-II, released from complex (cell wall) pectins by endo-polygalacturonase (Endo-PG) treatment, has been found to contain up to 15 (1→4)-linked-α-D-GalpA units, some of which carry four well-defined side chains, often referred to as A-, B-, C-, and D-side chains. Nevertheless, the relative locations on the backbone of these four branches, especially the A chain, remain to be ascertained. A combination of different data suggests that neither the terminal nonreducing GalA nor the contiguous GalA unit is likely to be the branching point of the A chain, but probably the ninth GalA residue from the reducing end, assuming a minimum backbone length of 11 (1→4)-linked-α-d-GalpA. The latest reports on RG-II are here highlighted, with a provided update for the macrostructure and array of functionalities.

scholarly journals Identification and characterization of a cell wall porin from Gordonia jacobaea

2017 ◽  
Vol 63 (5) ◽  
pp. 266-273
Author(s):  
Guadalupe Jiménez-Galisteo ◽  
Ester Fusté ◽  
Elisa Muñoz ◽  
Teresa Vinuesa ◽  
Tom G. Villa ◽  
...  
Keyword(s):  
Cell Wall ◽  
A Cell ◽  
Identification And Characterization

scholarly journals Identification and characterization of a cell-wall anchored DNase gene inClostridium perfringens

2005 ◽  
Vol 242 (2) ◽  
pp. 281-285 ◽  
Author(s):  
Kayo Okumura ◽  
Hameem I. Kawsar ◽  
Takeshi Shimizu ◽  
Toshiko Ohta ◽  
Hideo Hayashi ◽  
...  
Keyword(s):  
Cell Wall ◽  
A Cell ◽  
Identification And Characterization

scholarly journals Protocols for isolating and characterizing polysaccharides from plant cell walls: a case study using rhamnogalacturonan-II

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
William J. Barnes ◽  
Sabina Koj ◽  
Ian M. Black ◽  
Stephanie A. Archer-Hartmann ◽  
Parastoo Azadi ◽  
...  
Keyword(s):  
Cell Wall ◽  
Size Exclusion Chromatography ◽  
Plant Cell ◽  
Cell Walls ◽  
Size Exclusion ◽  
Proton Nuclear Magnetic Resonance ◽  
Cell Wall Polysaccharide ◽  
Rhamnogalacturonan Ii ◽  
Exclusion Chromatography

Abstract Background In plants, a large diversity of polysaccharides comprise the cell wall. Each major type of plant cell wall polysaccharide, including cellulose, hemicellulose, and pectin, has distinct structures and functions that contribute to wall mechanics and influence plant morphogenesis. In recent years, pectin valorization has attracted much attention due to its expanding roles in biomass deconstruction, food and material science, and environmental remediation. However, pectin utilization has been limited by our incomplete knowledge of its structure. Herein, we present a workflow of principles relevant for the characterization of polysaccharide primary structure using nature’s most complex polysaccharide, rhamnogalacturonan-II (RG-II), as a model. Results We outline how to isolate RG-II from celery and duckweed cell walls and from red wine using chemical or enzymatic treatments coupled with size-exclusion chromatography. From there, we applied mass spectrometry (MS)-based techniques to determine the glycosyl residue and linkage compositions of the intact RG-II and derived oligosaccharides including special considerations for labile monosaccharides. In doing so, we demonstrated that in the duckweed Wolffiella repanda the arabinopyranosyl (Arap) residue of side chain B is substituted at O-2 with rhamnose. We used electrospray-MS techniques to identify non-glycosyl modifications including methyl-ethers, methyl-esters, and acetyl-esters on RG-II-derived oligosaccharides. We then showed the utility of proton nuclear magnetic resonance spectroscopy (1H-NMR) to investigate the structure of intact RG-II and to complement the RG-II dimerization studies performed using size-exclusion chromatography. Conclusions The complexity of pectic polysaccharide structures has hampered efforts aimed at their valorization. In this work, we used RG-II as a model to demonstrate the steps necessary to isolate and characterize polysaccharides using chromatographic, MS, and NMR techniques. The principles can be applied to the characterization of other saccharide structures and will help inform researchers on how saccharide structure relates to functional properties in the future.

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