Design of high-throughput assays for the analysis of plant cell wall polysaccharides
Abstract: Plant cell walls contain a wide range of valuable polysaccharides possible to exploit for various purposes. To profile the diverse polysaccharides of plants, high-throughput, rapid and sustainable analytical methods are required. Currently, there are few methods designed for this purpose. An approach has been made to develop an analytical method based on enzymatic deconstruction and soft ionization mass spectrometry.Primary cell wall was extracted from the cells of the well-established model plant poplar to serve as an initial experimental subject. The cell wall was purified into alcohol insoluble residue consisting of only insoluble polysaccharides and a small amount of protein. Pure and specific enzymes were selected based on literature to deconstruct the polysaccharides and were evaluated on standard sugar substrates. Enzyme activity was measured using a rapid and reliable reducing sugar assay and achieved oligosaccharides were purified before electrospray ionization mass spectrometry. The outcome of the thesis presents an introduction to a high-throughput, rapid and sustainable analytical methodology for polysaccharide profiling in plant cell walls.
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