Radial flow chromatographic separation and purification method for biological polysaccharide

Abstract

The invention relates to a radial flow chromatographic separation and purification method for biological polysaccharide, which comprises the following steps of: centrifuging a medical fungi liquid fermentation product or a mycelium (fruiting body) enzymolysis product and collecting supernatant or collecting supernatant which comprises polysaccharide and is extracted from plants, animals, bacteriaand yeast; directly loading the supernatant on a macroporous resin radial chromatographic column of 500ml at the flow rate of 60 to 120ml per minute; eluting the macroporous resin radial chromatographic column by using distilled water or deionized water at the flow rate of 50 to 200ml per minute; and collecting 500 to 800ml of eluent, namely solution containing purified polysaccharide. The polysaccharide is directly separated and purified from the enzymolysis supernatant and the fermentation supernatant of the medical fungi mycelium and the supernatant extracted from the plants, animals, bacteria and yeast by a one-step method; and the biological polysaccharide separation and purification method has the advantages of simple process, convenient operation, no use of an organic solvent, highproduction efficiency and obvious purification effect.

Description

Technical field: [0001] The invention relates to technical fields such as separation science and engineering, biopharmaceuticals, etc., and specifically relates to a method for separating and purifying biological polysaccharides by utilizing macroporous resin radial flow chromatography technology. Background technique: [0002] Biological polysaccharides (including fungi, bacteria, plants, animals, etc.) have attracted extensive attention at home and abroad because of their remarkable anti-tumor, immune regulation, anti-oxidation, anti-aging and other biological activities. At present, most of the separation and purification strategies of biological polysaccharides are polysaccharide extraction supernatants are heated and concentrated—ethanol precipitation—centrifugation—to obtain crude polysaccharides—dialysis to remove small molecular impurities—hydrogen peroxide decolorization or activated carbon decolorization—Sevage method or trichloroacetic acid method to remove Miscel...

AI Technical Summary

Problems solved by technology

At present, most of the separation and purification strategies of biological polysaccharides are polysaccharide extraction supernatants are heated and concentrated—ethanol precipitation—centrifugation—to obtain crude polysaccharides—dialysis to remove small molecular impurities—hydrogen peroxide decolorization or activated carbon decolorization—Sevage method or trichloroacetic acid method to remove Miscellaneous protein—axial flow ion exchange chromatography or gel chromatography purification—eluent concentration—ethanol precipitation—centrifugation—precipitation and freeze drying—finally purified polysaccharides. It can be seen that the separation and purification process of traditional methods is complicated and the amount of chemical solvents large, poor environmental protection effect, low production efficiency, high production cost, and difficult to industrialized production. At the same time, due to the use of a large amount of organic solvents in the preparation process of polysaccharides, it not only causes harm to the operator and the environment, but also the residue of harmful substances affects the quality of the drug. Also cause a certain degree of adverse effects, therefore, the development of simple, efficient, clean, green separation and refining technology is a top priority

[0003] Radial flow chromatography technology is a new type of high-efficiency chromatography technology. Due to its radial flow principle, it can increase flow rate, reduce pressure, large capacity and load, easy to operate, and the chromatographic parameters obtained by laboratory or pilot test are stable. It can be directly applied to industrial production. At present, radial flow chromatography is mainly used to separate protein drugs. Unfortunately, there is no method for separating and purifying biological polysaccharides using radial flow chromatography.