| Catathelasma ventricosum (Peck) Sing is an edible fungi with high value of nutritionaland medicinal efficacy, which belongs to agaricales, tricholomataceae, pine of budmushroom genus. Now, the research of Catathelasma ventricosum (Peck) Sing focus on itsmonosaccharide composition and antidiabetic, antihyperglycemic, antihyperlipidemic andantioxidant activities. There are few studies on its Isolation, purification, structuralinvestigation and its other biological activities. The present research mainly studied theextraction, Isolation, purification, structural investigation and its antitumor activities ofpolysaccharides from Catathelasma ventricosum fruiting bodies. The main contents andconclutions are as follows:The contents of crude protein, polysaccharides, crude fiber, crude fat, ash, moisture，reducing sugar in the dry sample were:17.21%,52.48%,4.57%,1.77%,7.73%,10.61%,11.15%; the contents of mineral elements Mg, Mn, Fe, Zn were626.35mg/Kg,11.24mg/Kg,523.25mg/Kg,92.40mg/Kg, respectively. The total amino acids were17395.61mg/100g.The ultrasonic assisted extraction technology of CVP was investigated.one-factor-at-a-time and the orthogonal design mode experiment showed the optimumconditions were as follows: ultrasonic power70W, ultrasonic time10min, liquid-solid ratio20:1, extraction temperature60℃, and the yield could come to12.380%in the condition. Thefactors order affecting the yield of polysaccharides was: ultrasonic time>extractiontemperature>liquid-solid ratio>ultrasonic power.CVP was made after polysaccharide extracted from Catathelasma ventricosum fruitingbodies in the optimum conditions with a series of purification method. Its yield was1.3119%,pale yellow in colour. Five purified polysaccharides were obtained through DEAE-52ion-exchange column choromatography and Sephadex G-100column choromatography,which named CVP-Ⅰ, CVP-Ⅱ1, CVP-Ⅱ2, CVP-Ⅲ1, CVP-Ⅲ2. CVP-Ⅰ, CVP-Ⅱ2, CVP-Ⅲ1,CVP-Ⅲ2were identified as homogeneous components by means of GPC and Sephadex G-100column choromatography. The weight average relative molecular mass of CVP-Ⅰ, CVP-Ⅱ2,CVP-Ⅲ1, CVP-Ⅲ2were14331Da,11252Da,39359Da,13999Da. The physio-chemical properties of four kind of polysaccharides CVP-Ⅰ, CVP-Ⅱ2,CVP-Ⅲ1, CVP-Ⅲ2were explored. The contents of total sugar, protein, uronic acid and starchwere determined. CVP-Ⅰ, CVP-Ⅱ2, CVP-Ⅲ1, CVP-Ⅲ2were found to be non-starchpolysaccharides. The state and color of CVP-Ⅰ, CVP-Ⅱ2, CVP-Ⅲ1, CVP-Ⅲ2was white andpowder appearance, white and flocculent appearance, white and flocculent appearance, yellowand flocculent appearance, respectively; The total sugar of the four purified polysaccharideswas80.87%,86.76%,89.38%,85.78%, respectively; Uronic acid content of the four purifiedpolysaccharides was10.94%,12.93%,13.68%,12.93%, respectivly; Protein content of thefour purified polysaccharides was1.80%,0,0.51%,1.79%, respectively.The primary structure of CVPs was analysed by IR, GC, Periodate oxidation and Smithdegration, NMR.(1) The infrared spectra showed that CVP-Ⅰ, CVP-Ⅱ2, CVP-Ⅲ1, CVP-Ⅲ2had characteristic absorption peak of polysaccharide, because they contained the stretchingvibration of O-H at3428～3394cm-1, the stretching vibration of C-H at2929～2923cm-1, thestretching vibration of a group of carboxylic at1430～1200cm-1. The absorption peak at1645cm-1and1200～1000cm-1indicated that the four kinds of polysaccharides were pyranoidring and all had uronic acid,767cm-1and872cm-1characteristic absorption peak indicatedthe existence of mannose in CVP-Ⅰ.(2) Monosaccharide composition analysis of CVP-Ⅰ,CVP-Ⅱ2, CVP-Ⅲ1, CVP-Ⅲ2by GC showed that CVP-Ⅰ and CVP-Ⅱ2were both consistedof mannose, fucose, glucose and galactose, the mole ratios in CVP-Ⅰ wereMan:Fuc:Glu:Gla=1:10.66:10.33:34.26, the mole ratios in CVP-Ⅱ2were Man:Fuc:Glu:Gla=1:1.48:29.36:5.02; CVP-Ⅲ1was consisted of mannose and glucose, its mole ratios wereMan:Glu=1:41.27; CVP-Ⅲ2was consisted of mannose, glucose and galactose, its mole ratioswere Man:Glu:Gla=1:30.23:2.59;(3) Periodate oxidation and Smith degration results showedthat CVP-Ⅰ contained （1'）（,1'6）,（1'2）,（1'2,6）,(1'3),（1'2,3）（,1'3,6）,（1'2,4）,（1'2,3,4）type glycosidic bond, and the proportion of （1'）,（1'6）was58.20%, the proportion of（1'2）（,1'2,6）was9.03%, the proportion o（f1'2,3）（,1'3,6）,（1'2,4）,（1'2,3,4）was32.33%.(4)NMR results of C13and H1showed that CVP-Ⅰcontained、β-configuration and was consisited of4kinds of sugar residues, the main chainshould to be（1'6）type glycosidic bond. Antitumor activities of five kind of purified polysaccarides CVP-Ⅰ、CVP-Ⅱ1、CVP-Ⅱ2、CVP-Ⅲ1、CVP-Ⅲ2were tested on two kinds of cancer cells, the human breastcancer cells MCF-7and the human liver cancer cell line BEL-7402by MTT assay. Theresults showed that CVP-Ⅰ, CVP-Ⅱ1, CVP-Ⅱ2, CVP-Ⅲ1, CVP-Ⅲ2all had certain inhibitionto this two kinds of tumor cells，presented dose-dependent and time-dependent manner. Theinhibitory effect of purified polysaccharides CVP-Ⅲ1、CVP-Ⅲ2were both very obvious. |
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