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Purification of water soluble bioactive polysaccharides from biomass of heterotrophic mutant alga Chlorella vulgaris G11.
Sushytskyi, L. ; Lukáč, Pavol ; Bleha, R. ; Čopíková, J. ; Vannucci, Luca ; Sinica, A. ; Kaštánek, P.
Water extract obtained from biomass of mutant alga Chlorella vulgaris G11 was deproteinized and then purified with AEC on DEAE Sepharose FF and SEC on Sepaphadex G-75 superfine columns. Two high MW (>1000 kDa and 98,3 kDa) polysaccharides (CPS1 and CPS3) were eluted with water and 0.5M NaCl on DEAE column, and heteropolysaccharide with MW of 15 kDa (CPS2) was also eluted with water on DEAE. The former fractions were identified as branched rhamnans, the latter one as branched xylogalactofuranan of unusual structure. CPS1 shown properties of unstable polimer: according to GPC analysis and by using light scattering and RI detectors it was found, that some fraction of polymers (260-800 kDa) have self-assembling properties in aqueous solution, and MW of this fraction decreased during some time while polimer with initially bigger MW (>1000 kDa) became bigger. CPS3 was eluted with 0.3-0.5 M of NaCl on DEAE Sepharose FF column and have MW of 98,3 kDa. Cell toxicity of the products were tested in vitro on cell cultures for different concentrations
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