Colletotrichum gloeosporioides biomass was submitted to ethanolic extraction (78 ºC, 12 h) to remove low molecular weight compounds followed by hot aqueous extraction (100 ºC, 4 h, 4x) to obtain polysaccharides. Consecutive freezing/thawing treatments of the aqueous extract yielded a soluble fraction in cold water that was separated on five components (PI to PV) by gel exclusion chromatography on Sepharose CL-6B. The PIV was homogeneous on HPSEC/RID, with apparent MW of 2.8 x 104 g mol-1, Mw/Mn 1.17, 100% of total sugar. The PIV was composed by galactose (61%), glucose (34%) and mannose (5%). 1H and 13C NMR analysis mono- and two-dimensional, homo- (COSY) and heteronuclear (HSQC and HMBC) showed that PIV is a heteropolysacharide with a main chain constituted by β-D-galactofuranosidic units (1→5) and (1→6) linked with some of the β-D-galactofuranosidic (1→6) units substituted in O-3 by α-D-glucopiranosidic residues. Cytotoxic assays (MTT) indicated that the viability of CHO-K1 cells, when exposed to different heteropolysaccharide concentrations, did not show significant differences when compared to the viability of the negative control (CN), in the experimental times of 24 and 48 hours. These results suggest the possibility of new cytotoxic assays in other normal cell lines as well as in tumor cells.
Keywords:
heteropolysaccharide; HPSEC/RID; HSQC; MTT assay
