Isolation,Purification,Structural Elucidation And Biological Activities Of Polysaccharide From Camphoratum(Bull.) Fr.Lactarius

Lactarius camphorates(Bull.) Fr. is a kind of fungi belonging to Agaricales, Russulaceae, Lactarius which is not only a delicious edible and medieinal fungus, but also has strong antibacterial activity and anti-oxidation, anti-aging features. It is an ideal natural flavor source and food additives. At present, researches on L. camphorates(Bull.) Fr. were focused on the planting method, nutritional element and antibacterial activity and anti-oxidation activities of the mycelial extracts. Researches on the homogenous polysaccharide from L. Camphorates(Bull.) Fr. are few reports. First, we extracted polysaccharides with boiling water and purified by DEAE-cellulose column. Then the structures of LC-1 were elucided by chemical(methylation analysis, partial acid hydrolysis,ete.)and spectroscopic methods(1D NMR:1HNMR,13CNMR). Last, using biochemical methods and modern biotechnolog method to reseach anti-oxidant and anti-tumor activities of polysaccharides from the fruiting bodies of L.camphoratum(Bull.) Fr.. it is an essential prerequisite for research of the mechanism of anti-tumor activities and immunodulatory activities of LC-1, and it can provide scientific basis for research and development of polysaccharide-targeted immune regulation.This dissertation consists of three chapter, which are extraction、purification and fractionation of polysaccharides from L.camphoratum(Bull.) Fr., chemical structural analysis, anti-oxidant and anti-tumor activities of polysaccharides from the fruiting bodies of L.camphoratum(Bull.) Fr.. 1 Studies on isolation, purification and fractionation of polysaccharides from L.camphoratum(Bull.) Fr.The fruiting bodies of L. camphoratum(Bull.) Fr. were soaked with 95% EtOH, followed by extracted with boiling water and precipitated crude polysaccharides yield of 10.2%. And then purified using DEAE-cellulose column, one water-soluble homogeneous polysaccharides(LC-1) were obtained. Lack of absorption at 280 nm and 260 nm by UV scanning indicated that it contained no protein and nucleic acid. IR spectra showed that the sugar characteristic groups were existed. High performance gel permeation chromatography(HPGPC) producing a single symmetrical peak, indicated that it was homogeneous and it molecular weight was around 9279 Da. 2 Structure elucidation of LC-1The structures of LC-1 were elucided by chemical(methylation analysis, partial acid hydrolysis,ete.) and spectroscopic methods(1D NMR:1HNMR,13CNMR), the number of sugar residues, the form of glycosyl linkage, the sequence of sugar residues, the configuration of anomeric center. It is concluded that LC-1 contained D-Glu and D-Gal in the ratio of 2:1; a repeating unit of LC-1 has a backbone of(1�'4)--dglucosepyranose which branches at O-6 based on the experimental results. The branch was supposed to be the composition of one with�'3)--d-galactopyranose residue. 3 Antioxidant activities and anti-tumor activities in vitro of LC-1The antioxidant activities in vitro of LC-1 were evaluated by measuring 2,2-dipheny 1-1-picrylhydrazyl(DPPH-) radical and ABTS+ radical scavenging activity. The results showed that the LC-1 had antioxidant activities with a dose-dependent in the testing dose range. CCK-8 assay was used to investigate the anti-proliferation activities against cervical cancer cells(HeLa) in vitro of LC-1. The results showed that the LC-1 had significantly toxicity effects on cervical cancer HeLa cell line, It can significantly inhibite HeLa’s proliferation in dose-dependent and time-dependent manner.