The dissociation/deconstruction of the different components is a prerequisite for the analysis of ex-situ cell wall components. CD BioSciences offers our customers a variety of chemical, enzymatic, and physical methods to accomplish linkage breakdown between polymers. We aim to release all cell wall components without altering their chemical composition.

Introduction

Plant cell walls are complex 3D networks composed of proteins, polysaccharides, and lignin. Multiple molecular interactions make the analysis of cell wall compounds a more complex task. In order to analyze ex-situ cell wall components, their destructuration /dissociation/disorganization from each other is a prerequisite for their analysis. The dissociation of plant cell walls depends heavily on the number and type of bonds (covalent, electrostatic, hydrogen, ionic linkage) between them. The disintegration of bonds between these polymers can be achieved by chemical, physical, enzymatic, or by a mixture of two or three of these methods. Developing efficient and rapid methods to release all cell wall components without altering their chemical compositions is currently a major challenge in the in vitro analysis of cell wall components.

Fig.1 Diagrammatic representation of the major structural polysaccharide components of a "typical" primary plant cell wall. (Flint, 2012)

Our Services

Researchers at CD BioSciences have developed a variety of methods for purifying plant cell wall polymers to help clients characterize cell wall composition and to lay the experimental foundation for understanding the function of different cell wall polymers in plant development and adaptation. Our services include but are not limited to:

• Protein purification
  Plant cell walls contain two groups of cell wall proteins (CWP), including proteins formed with amino acids only and glycoproteins with a non-amino acid component. Depending on the strength of their attachment to the wall, we adopt different strategies for the separation of labile and weakly bound proteins. For example, we extract weakly bound proteins (hydrogen bonds, van der Waals interactions, hydrophobic or ionic interactions) by salt solutions (CaCl₂, LiCl, NaCl).
• Polysaccharide purification
  We offer three methods for the purification of plant cell wall polysaccharides. The first is the partial purification of one family of polysaccharides without focusing on other cell wall components. The second is the extraction of polysaccharides by a sequential extraction procedure, which is suitable for collecting a large number of fractions enriched in one compound. The last one uses chemical or physical processes suitable for solubilizing cell wall components.
• Cellulose isolation
• Pectins isolation
• Hemicelluloses isolation
• Lignin purification
  Acidolysis and enzymatic methods are the two main approaches for lignin isolation. We combined these two methods, i.e. cellulolytic enzyme treatment followed by mild acid digestion, to obtain higher lignin yields with fewer impurities.
• Cell wall destructuration
  Biomass disruption is another strategy for the analysis of plant cell wall composition. We have developed methods of chemical dissociation and physical dissociation (such as comminution, steam explosion and hydrothermolysis) to breakdown polymer linkages, resulting in complete cell wall disintegration. The free polymer can then be dissolved prior to analysis.

With a specialized biophysical analysis platform, CD BioSciences offers a full range of cell wall components characterization services, including pre-analytical cell wall component extraction. If you need our technical support, please contact us and we will be happy to develop the best experimental protocol for you.

References

1. Flint, H. J.; et al. Microbial degradation of complex carbohydrates in the gut. Gut microbes. 2012, 3(4): 289-306.
2. Costa, G.; Plazanet, I. Plant cell wall, a challenge for its characterisation. Advances in biological chemistry. 2016, 6(3):2162-2183.

• In Vitro Plant Cell Wall Components Analysis
• In Situ Plant Cell Wall Components Analysis