Regulation On Synthesis Of Polysaccharide In Glycyrrhiza Inflata Bat. Callus And Biological Activity Analysis

As one of the most crucial Chinese traditional medicine, the radices and rhizomes ofGlycyrrhiza spp. containing triterpene glycosides, compounds such as flavonoids andpolysaccharides, which grows in arid and semi-arid desert steppe, desert margins and loesshilly area. As an important biological activity of components in Glycyrrhiza spp, Glycyrrhizapolysaccharides （GPS） can regulate the body’s cell division or differentiation and growth oraging, as well as have a variety of immune function and the role of strong anti-tumor virusresistance. Modern researches of chemical and biological activity of Glycyrrhiza spp. aremostly focused on Glycyrrhizic acid and flavonoids, but lack of the research reports ofglycyrrhiza polysaccharides. This paper to one of the three major varieties of licoriceresources of Glycyrrhiza inflate Bat. for the study, MS medium on the basis of, by adjustmentmedium of nutrition components on expansion fruit licorice more injury organization cell ofgrowth for regulation, achieved the purpose which has increased GPS in its product. Andusing orthogonal experimental method to find out the ultrasonic extraction of callus tissue ofglycyrrhiza polysaccharide in optimal conditions.By protein clearing, and semi-permeable membrane dialysis, and chromatographycolumn washing, most of the impurities has been removed from GPS. Finaly, we comparedthe GPS that extracted from Callus with GPS in the same conditions of extraction andpurification of natural licorice medicinal materials which have anti-oxidant and anti-tumoractivity.Results show that:a.Among the tested basal medium, MS with the addition of sucrose was the best forcallus growth,6,7-V was most suitable for glycyrrhiza polysaccharide production. Theoptimal concentration of PO₄^3–and Ca²⁺of medium was1.25mmol/L and2.99mmol/L forbiomass accumulation of callus and bio-synthesis of glycyrrhiza polysaccharide. The higherconcentration of phosphate and Ca²⁺were benefit of the production of glycyrrhizapolysaccharide, but not the growth of callus.b.The extraction rate of polysaccharide from Glycyrrhiza inflata callus induced from cotyledon reached the maximum content of7.72%and when the liquid-solid ratio was70:1,ultrasonic power with80W,90℃and30min, content of Glycyrrhiza polysaccharide was86.7980%. Extraction rate of polysaccharide in Glycyrrhiza inflata callus from hypocotylreached the maximum content of7.49%,and when the liquid-solid ratio was40:1,afterultrasound with80W,80℃a nd20min, content ofGlycyrrhiza polysaccharide was76.7110%.The extraction rate of polysaccharide from medicinal materials was only5.26%, and the ofGlycyrrhiza polysaccharide was only58.374%, which is significantly lower than thepolysaccharide from Glycyrrhiza inflata callus.c.We use the method of Sevag flowing water dialysis DEAE-52and Sephadex G-75tothe further purify GPS. The content of polysaccharide in Glycyrrhiza inflata callus were92.685%, which is clearly higher than the polysaccharide in medicinal materials86.424%.d.Glycyrrhiza polysaccharides had a certain degree of anti-oxidant and anti-tumoractivity, and the effect of scavenging free radicals and inhibiting tumor cells of thepolysaccharide from Glycyrrhiza inflata callus were more visible than from medicinalmaterials.At present most of glycyrrhiza polysaccharide obtained based on the natural licoriceherbs, but because of over-exploitation of resources causing scarce. In the experiment, weextract GPS by inducting callus of glycyrrhiza and from which extracts on tissue culture cellsof glycyrrhiza polysaccharides constituents, bio-activity of polysaccharides from medicinalplants and natural licorice made a preliminary study of the similarities and differences.Theexperiment results show that, through the expansion of Glycyrrhiza inflata Batal callusgrowth and polysaccharides synthesis regulation of medium can be given more GPS thanafter using MS basic of glycyrrhiza polysaccharide. The GPS which were obtained throughtissue culture methods and were in natural licorice herbal medicine have a certainanti-oxidant and anti-tumor activity, and GPS in the callus have an advantage. The researchhas opened up new avenues for the large-scale production of glycyrrhiza polysaccharide.