Purification And Physicochemical Properties Of The D-grifolan Component

Systematic studies on the extraction , purification and structural characterization of Grifola frondosa polysaccharides and its D-fraction were carried out in the presented dissertation. Some new methods and points were developed on the basis of thesis of these studies. The results and creative point are described as follows:1. Found the content of polysaccharides in the crude polysaccharides is 28.9%,and the content of protein is 35.61% by Kjeldahl determination, and the content of 3 -glucan is 3.3%~3.4%.2. Built a simple detecting method about G-D by HPLC According to some scientific reports and the analysis of the products on the market which are described mainly constituted with G-D.3. The optimal concentration of ethanol are 45% and 80% in the two alcohol -insoluble stages; DEAE Sepharose Fast Flow is suitable for G-D ion exchange chromatography with the eluant at pH 7.2 and Sepheracyl S-400 is appropriate for the further purification of G-D with 1mol/L NaCl eluant at pH 7.2. G-D is a brownish black , deliquescent and diffluent powder ,whose specific rotatory power is -62. 5 . Found the content of polysaccharides in G-D is 79.5%,and the content of protein is 16.2%.4. Using papain, get G-D glycopeptide. By β -elimination and hydrazinolysis,found there is no or a few 0-type glycopeptide bond but N-type glycopeptide bond in G-D. After degraded by H2SO4 there are three kinds of monosaccharide: D-glucose, D-galactose, D-mannitose,and the ratio isl.8:0.4: 29.4. The FT-RI and 1H ,13C NMR spectrum indicated that there are β -(1â†'6)-D-mannopyranose in G-D chains.5.By compared the content of G-D in the Grifola frondosa crude polysaccharides from Qingyuan and Qianxi, find that which in the former is 1.78 times as in the later.6.Adopting FPLC to separate other glucoproteins in the crude polysaccharides can make every component reach the separation of the certain degree. But the type of FPLC columns and the elution should be further optimized.the innovations that originally studies is embodied in the following several points:1. A simple , convenient and practical detection method of G-D have been set up.2. A simple and convenient, reliable extraction and purification method of G-D have been established3. The way of joining between polysaccharide chains and peptide chains in G-D have been analysed, confirm that it may contain a few O-type glucoprotein, but must contain N-type glucoprotein.4. The monoses of G-D were analysed ,it contains outside a large amount of mannose , A small amount of glucose, galactose exist yet.5. the units of G-D chains was detected, it mostly contains β -(1â†' 6)-D-mannopyranose .