A study of the molecular weight distribution of natural polysaccharides and polyacetylenes by means of multidimensional chromatography
Abstract
The molecular-weight and functional characteristics of the products of the biosynthesis of polysaccharide and polyacetylene compounds by basidiomycetes genus Ganoderma lucidum and Pleurotus ostreatus were studied using multidimensional liquid chromatography together with gel permeation chromatography and reversed-phase adsorption chromatography. The fungi were grown by means of submerged cultivation in a medium consisting of wheat bran on the Chapek–Doks medium of the following composition (g/l): NaNO3 -2, K2HPO4 -1, MgSO4·7H2O-0.5, KCl-0.5, FeSO4·H2O-0.01, glucose-30%, and H2O. The cultivation was performed in a thermostat at 2800 С. After 10 days of exposure, polysaccharides and polyacetylene compounds were found in the fungi mycelium. The resulting fungal biomass was then ground and extracted with a methyl ether of acetic acid and water. The precipitate and the extract were analysed separately using the said chromatography system. The analysis demonstrated that the water extract contained 70-80% of polysaccharides, and the ether extract contained 65-70% of polyacetylene compounds. Thus, the GPC demonstrated that the products of biosynthesis contained 5 fractions, including 4 fractions of polysaccharides, consisting of hexaose (Мw=Mn=1085), heptaose (Мw=Mn=1260) and two oligosaccharides with Мw and Mn 4390, 3600 and 5450, 4500, respectively, and one polyacetylene fraction (Мw =180, Mn =150). The reversed-phase adsorption chromatography with UV-detection demonstrated that the polyacetylene fraction contained 5 subfractions, which by their chemical nature are related to polyacetylene acids and alcohols conjugated by various combinations of double and triple bonds. In order to study the chemical composition of the detected subfractions, the polyacetylene fraction was subjected to preparative separation. The separation was performed with increased extract concentration by means of multiple fraction collection according to the corresponding peaks on the LSC chromatogram. The peaks were then subjected to IR spectroscopy, which allowed us to determine their functional composition. The composition of the polyacetylene compounds was also confirmed chromatographically by the retention volumes of the tested compounds using the corresponding standards.
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References
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