Monoclonal Antibodies, Carbohydrate-Binding Modules, and Detection of Polysaccharides in Cell Walls from Plants and Marine Algae

Author(s):  
Delphine Duffieux ◽  
Susan E. Marcus ◽  
J. Paul Knox ◽  
Cécile Hervé
BioResources ◽  
2007 ◽  
Vol 2 (2) ◽  
pp. 284-295
Author(s):  
Clemens Altaner ◽  
J. Paul Knox ◽  
Michael C. Jarvis

Wood cell wall polysaccharides can be probed with monoclonal antibodies and carbohydrate-binding modules (CBMs). Binding of monoclonal antibodies to β-1-4-xylan, β-1-4-mannan, β-1-3-glucan, and α-1-5-arabinan structures were observed in native Sitka spruce (Picea sitchensis (Bong.) Carrière) wood cell walls. Furthermore CBMs of different families, differing in their affinities for crystalline cellulose (3a) and amorphous cellulose (17 and 28), were shown to bind to the native wood cell walls with varying intensities. Resin channel forming cells exhibited an increased β-1-4-xylan and a decreased β-1-4-mannan content. Focusing on severe compression wood (CW) tracheids, β-1-3-glucan was found towards the cell lumen. In contrast, α-1-5-arabinan structures were present in the intercellular spaces between the round tracheids in severe CW, highlighting the importance of this polymer in cell adhesion.


2011 ◽  
Vol 77 (20) ◽  
pp. 7217-7226 ◽  
Author(s):  
Sabine Gruber ◽  
Christian P. Kubicek ◽  
Verena Seidl-Seiboth

ABSTRACTMycoparasiticTrichodermaspecies have expanded numbers of fungal subgroup C chitinases that contain multiple carbohydrate binding modules and could thus be important for fungal cell wall degradation during the mycoparasitic attack. In this study, we analyzed the gene regulation of subgroup C chitinases in the mycoparasiteTrichoderma virens. In addition to regulation by nutritional stimuli, we found complex expression patterns in different parts of the fungal colony, and also, the mode of cultivation strongly influenced subgroup C chitinase transcript levels. Thus, the regulation of these genes is governed by a combination of colony-internal and -external signals. Our results showed completely different expression profiles of subgroup C chitinase genes inT. virensthan in a previous study withT. atroviride, although both fungi are potent mycoparasites. Only a few subgroup C chitinase orthologues were found inT. atrovirideandT. virens, and even those showed substantially divergent gene expression patterns. Microscopic analysis revealed morphogenetic differences betweenT. atrovirideandT. virens, which could be connected to differential subgroup C chitinase gene expression. The biological function of fungal subgroup C chitinases therefore might not be as clear-cut as previously anticipated. They could have pleiotropic roles and might be involved in both degradation of exogenous chitinous carbon sources, including other fungal cell walls, and recycling of their own cell walls during hyphal development and colony formation.


2006 ◽  
Vol 103 (12) ◽  
pp. 4765-4770 ◽  
Author(s):  
L. McCartney ◽  
A. W. Blake ◽  
J. Flint ◽  
D. N. Bolam ◽  
A. B. Boraston ◽  
...  

2021 ◽  
pp. 100638
Author(s):  
Marie Sofie Møller ◽  
Souad El Bouaballati ◽  
Bernard Henrissat ◽  
Birte Svensson

Materials ◽  
2021 ◽  
Vol 14 (12) ◽  
pp. 3175
Author(s):  
Mariana Barbosa ◽  
Hélvio Simões ◽  
Duarte Miguel F. Prazeres

Materials with novel and enhanced functionalities can be obtained by modifying cellulose with a range of biomolecules. This functionalization can deliver tailored cellulose-based materials with enhanced physical and chemical properties and control of biological interactions that match specific applications. One of the foundations for the success of such biomaterials is to efficiently control the capacity to combine relevant biomolecules into cellulose materials in such a way that the desired functionality is attained. In this context, our main goal was to develop bi-functional biomolecular constructs for the precise modification of cellulose hydrogels with bioactive molecules of interest. The main idea was to use biomolecular engineering techniques to generate and purify different recombinant fusions of carbohydrate binding modules (CBMs) with significant biological entities. Specifically, CBM-based fusions were designed to enable the bridging of proteins or oligonucleotides with cellulose hydrogels. The work focused on constructs that combine a family 3 CBM derived from the cellulosomal-scaffolding protein A from Clostridium thermocellum (CBM3) with the following: (i) an N-terminal green fluorescent protein (GFP) domain (GFP-CBM3); (ii) a double Z domain that recognizes IgG antibodies; and (iii) a C-terminal cysteine (CBM3C). The ability of the CBM fusions to bind and/or anchor their counterparts onto the surface of cellulose hydrogels was evaluated with pull-down assays. Capture of GFP-CBM3 by cellulose was first demonstrated qualitatively by fluorescence microscopy. The binding of the fusion proteins, the capture of antibodies (by ZZ-CBM3), and the grafting of an oligonucleotide (to CBM3C) were successfully demonstrated. The bioactive cellulose platform described here enables the precise anchoring of different biomolecules onto cellulose hydrogels and could contribute significatively to the development of advanced medical diagnostic sensors or specialized biomaterials, among others.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hong-Hsing Liu ◽  
Yu-Chen Lin ◽  
Chen-Shuan Chung ◽  
Kevin Liu ◽  
Ya-Hui Chang ◽  
...  

AbstractBowel microbiota is a “metaorgan” of metabolisms on which quantitative readouts must be performed before interventions can be introduced and evaluated. The study of the effects of probiotic Clostridium butyricum MIYAIRI 588 (CBM588) on intestine transplantees indicated an increased percentage of the “other glycan degradation” pathway in 16S-rRNA-inferred metagenomes. To verify the prediction, a scoring system of carbohydrate metabolisms derived from shotgun metagenomes was developed using hidden Markov models. A significant correlation (R = 0.9, p < 0.015) between both modalities was demonstrated. An independent validation revealed a strong complementarity (R = −0.97, p < 0.002) between the scores and the abundance of “glycogen degradation” in bacteria communities. On applying the system to bacteria genomes, CBM588 had only 1 match and ranked higher than the other 8 bacteria evaluated. The gram-stain properties were significantly correlated to the scores (p < 5 × 10−4). The distributions of the scored protein domains indicated that CBM588 had a considerably higher (p < 10−5) proportion of carbohydrate-binding modules than other bacteria, which suggested the superior ability of CBM588 to access carbohydrates as a metabolic driver to the bowel microbiome. These results demonstrated the use of integrated counts of protein domains as a feasible readout for metabolic potential within bacteria genomes and human metagenomes.


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