Isolation of protein-polysaccharide conjugates from Solanum tuberosum L. rootlets by extended reversed-phase flash chromatography
Аннотация
Polysaccharides and their conjugates with various biologically active compounds (proteins, nucleic acids, peptides etc.) are of great importance, they take a part in various biological processes. The one of the important feature of polysaccharides is possibility to form wide variety of branched structures with different molecular masses and, thus, they can form very complicated structural conjugates with wide range of biologically active compounds. Cells of tubers of cultivated potato (Solanum tuberosum L.) contain high amounts of polysaccharides conjugated with proteins and nucleotides. In this work we developed fast and efficient chromatographic procedure for the isolation of protein-polysaccharide conjugates from the rootlets of Solanum tuberosum L. It consisted of double-cartridge flash-chromatographic system, filled with the cross-linked macroporous styrene-divinylbenzene and C18 stationary phases. Isolated products were characterized by different analytical methods, including HPLC (reversed phase and size exclusion chromatography), gas chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (determination of proteins). Some structural features were determined by means of IR spectroscopy in the region characteristic for saccharides (1250-750 cm-1). It could be concluded, that isolated conjugates consisted of arabinogalactane polysaccharide and protein with average MM 60 kDa. Analytical data obtained for both products, isolated using conventional precipitation method and flash-chromatographic procedure were similar.
Скачивания
Литература
Wasser S.P. Medicinal mushrooms as a source of antitumor and immunomodulat-ing polysaccharides. Applied Microbiology and Biotechnology. 2002; 60: 258-274. https://doi.org/10.1007/s00253-002-1076-7
Zong A., Cao H., Wang F. Anticancer polysaccharides from natural resources: A review of recent research. Carbohydrate Polymers. 2012; 90: 1395-1410. https://doi.org/10.1016/j.carbpol.2012.07.026
Furrer A., Cladis D.P., Kurilich A., Manoharan R., Ferruzzi M.G. Changes in phenolic content of commercial potato va-rieties through industrial processing and fresh preparation. Food Chem. 2017; 218: 47-55. https://doi.org/10.1016/j.foodchem.2016.08.126
Aversano R., Contaldi F., Adelfi M.G., D’Amelia V., Diretto G., De Tommasi N., Vaccaro C., Vassallo A., Carputo D. Com-parative metabolite and genome analysis of tuber-bearing potato species. Phytochemis-try. 2017; 137: 42-51. https://doi.org/10.1016/j.phytochem.2017.02.011
Zhang Q.-W., Lin L.-G., Ye W.-C. Techniques for extraction and isolation of natural products: a comprehensive review. Chinese Medicine. 2018; 13: 1-26. http://dx.doi.org/10.1186/s13020-018-0177-x
Reid R.G., Sarker S.D. Isolation of Natural Products by Low-Pressure Column Chromatography. Natural Products Isola-tion. 2012; 864: 155-187. https://doi.org/10.1007/978-1-61779-624-1_7
Shi L. Bioactivities, isolation and puri-fication methods of polysaccharides from natural products: A review. International Journal of Biological Macromolecules. 2016; 92: 37-48. https://doi.org/10.1016/j.ijbiomac.2016.06.100
Ding X., Zhu F., Gao S. Purification, antitumour and immunomodulatory activi-ty of water-extractable and alkali-extractable polysaccharides from Solanum nigrum L. Food Chem. 2012; 131; 677-684. https://doi.org/10.1016/J.FOODCHEM.2011.09.060
Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T44. Nature. 1970; 227: 680-685. https://doi.org/10.1038/227680a0
Ranathunge K., Schreiber L., Franke R. Suberin research in the genomics era – New interest for an old polymer. Plant Science. 2011; 180: 399-413. https://doi.org/10.1016/j.plantsci.2010.11.003
Synytsya A., Novak M. Structural analysis of glucans. Ann. Transl. Med. 2014; 2: 17. https://doi.org/10.3978/j.issn.2305-5839.2014.02.07
Kaсurakova M., Capek P., Sasinko-va V., Wellner N., Ebringerova A. FT-IR study of plant cell wall model compounds: pectic polysaccharides and hemicelluloses. Carbohydrate Polymers. 2000; 43: 195-203. http://dx.doi.org/10.1016/S0144-8617(00)00151-X