Array-based microfibril surface assessment (AMSA): a method for probing surface-exposed polysaccharides on cellulose nan
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ORIGINAL RESEARCH
Array-based microfibril surface assessment (AMSA): a method for probing surface-exposed polysaccharides on cellulose nanofibres Alixander Perzon Peter Ulvskov
. Stjepan Kresˇimir Kracˇun
. Bodil Jørgensen
.
Received: 24 May 2020 / Accepted: 10 August 2020 Ó Springer Nature B.V. 2020
Abstract Small amounts of polysaccharides remaining tightly bound to cellulose microfibrils in the plant cell wall are significant to cell wall architecture and of importance to the rheological properties of plantderived cellulose nanofibres (CNFs). A method for detecting these polysaccharides is presented. The
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10570-020-03398-x) contains supplementary material, which is available to authorized users. A. Perzon S. K. Kracˇun B. Jørgensen P. Ulvskov (&) Department of Plant and Environmental Sciences, Section for Glycobiology, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark e-mail: [email protected] A. Perzon e-mail: [email protected]
method, Array-based Microfibril Surface Assessment (AMSA), uses monoclonal antibodies to probe polysaccharides that are exposed on fibrous material. Microfluidised alcohol insoluble residue from sugar beet pulp was used as control and baseline, representing a minimally processed material. To test the utility of AMSA, CNF prepared from sugar beet pulp were analysed. Surprisingly, nanofibrillation did not enhance binding of CBM3a (specific to crystalline cellulose) which indicates that bare cellulose is not easily made. However, nanofibrillation exposed a large amount of unbranched mannan, implying that mannan is closely associated with cellulose in these cell walls. Additional treatment of the CNF suspensions with pectinases removed the majority of homogalacturonan and rhamnogalacturonan I, which caused flocculation of the samples. This highlights the importance of preserving small quantities of these polysaccharides to promote CNF-water interactions and obtain well-dispersed fibres. In conclusion, it is demonstrated how AMSA has a utility in both biotechnological applications involving CNF as well as for exploring plant cell wall architecture.
S. K. Kracˇun e-mail: [email protected] B. Jørgensen e-mail: [email protected] Present Address: S. K. Kracˇun Chr. Hansen A/S, Discovery R&D, Microbial Screening Department, Bøge Alle 10-12, 2970 Hørsholm, Denmark
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Cellulose
Graphic abstract
Substrate Insoluble dye product
Robot spots cellulose nanofiber suspensions onto nitrocellulose membrane Probing with primary, glycan-specific antibody Develop using enzymeconjugated secondary antibody Keywords Cellulose Surface Polysaccharides Nanofibers Microfibrils AMSA
Introduction The primary plant cell wall confers strength and is essential for growth and development of plants (Caffall and Mohnen 2009). The polysaccharide matrix that shapes the primary cell wall involves cellulose, hemicelluloses, and pectin.
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