Characterization and immunostimulating activity of a water-soluble polysaccharide isolated from <Emphasis Type="Italic">Haematococcus lacustris</Emphasis>

A water-soluble polysaccharide was isolated and purified from the culture filtrate of the photosynthetic green microalgae Haematococcus lacustris by 75% ethanol precipitation and Sepharose CL-6B column chromatography. The molecular mass of the purified polysaccharide (named HCP) was estimated to be approximately 135 kDa by size-exclusion HPLC and its monosaccharide composition was galactose, glucose and mannose at a relative molar ratio of 2.0, 1.0, and 4.1, respectively, suggesting that HCP is a galactomannan. Fourier-transform infrared and elemental analysis revealed that the purified HCP contains sulfate esters by 1.08% (in mass) and no detectable level of protein. The HCP significantly stimulated murine macrophage RAW264.7 cells to secrete the pro-inflammatory cytokine, TNF-α, in a dose-dependent manner and also enhanced the expression of COX-2 and iNOS genes at a concentration of lower than 10 μg/mL HCP. These results indicated that the sulfated HCP of H. lacustris has potent early innate immune stimulating activities.