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Method for modifying functional plants polysaccharide

A modification method, plant polysaccharide technology, applied in the field of functional plant polysaccharide modification, can solve the problems of low toxicity and low transfection efficiency, and achieve high recovery rate and high gene transfection efficiency

Inactive Publication Date: 2008-08-06
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Non-viral gene carriers have problems such as toxicity, biocompatibility, and degradability. At present, there is a synthetic material polyethyleneimine (PEI). Experiments have found that polyethyleneimine with a molecular weight of 22000-25000 has a very high transfection efficiency. However, there are problems of toxicity and degradability. Polyethyleneimine with small molecular weight (600, 1200, 2000) has low toxicity and is easy to metabolize in vivo, but has almost no transfection efficiency

Method used

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  • Method for modifying functional plants polysaccharide
  • Method for modifying functional plants polysaccharide
  • Method for modifying functional plants polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1) Polysaccharide extraction and purification

[0033] Take 500 grams of any of biologically active medicinal plants, such as shiitake mushrooms, scutellaria baicalensis, ganoderma lucidum, rice kernels, cactus, versicolor versicolor, and wolfberry, add 3 times the amount of water and boil for 3 hours, and add the obtained solution to 95% ethanol solution after concentration , stirring while adding, so that the final ethanol concentration was 65%, after centrifugation, the resulting precipitate was freeze-dried to obtain 22.5 grams of crude polysaccharide.

[0034] Take 10 grams of crude polysaccharide, dissolve it in 200 ml of aqueous solution, put it into a dialysis bag with a molecular weight of 15,000 and dialyze for 48 hours after dissolving, take out the solution in the bag, add 2 grams of activated carbon for decolorization, filter, and continue to use the filtrate with a dialysis bag with a molecular weight of 15,000 After dialysis for 48 hours, the solution in ...

Embodiment 2

[0042] Take 0.5 g of the purified Scutellaria baicalensis polysaccharide in Example 1 and dissolve it in 15 ml of phosphate buffer. Take 0.2 g of N,N'-disuccinimidyl sulfate, dissolve it in 10 ml of dichloromethane, and slowly add it to the polysaccharide solution under the protection of nitrogen and avoiding light, and stir while adding. After the internal addition is completed, react in the dark for 90 minutes after the addition.

[0043] Take 0.6 g of polyethyleneimine with a molecular weight of 1200, dissolve it in 15 ml of phosphate buffer solution, add 0.2 ml of triethylamine as a catalyst, add it into the activated polysaccharide solution under the protection of light and nitrogen, and stir while adding, Complete the addition within 150 minutes, and react in the dark for 150 minutes after the addition.

[0044] The completed solution was put into a dialysis bag with a molecular weight cut off of 15,000, dialyzed for 48 hours, and the solution in the bag was freeze-drie...

Embodiment 3

[0047] Take 0.1 g of Polysaccharide polysaccharide purified in Example 1 and dissolve it in 5 ml of phosphate buffer. Take 0.3 g of benzotriazole carbonate, dissolve it in 10 ml of dichloromethane, and slowly add it to the polysaccharide solution under nitrogen protection and dark conditions, do not add while stirring, add it within 100 minutes, after the addition is complete React for 80 minutes.

[0048] Take 0.4 g of polyethyleneimine with a molecular weight of 2000, dissolve it in 10 ml of phosphate buffer solution, add 0.3 ml of triethylamine as a catalyst, add it into the activated polysaccharide solution under the protection of light and nitrogen, and stir while adding, Complete the addition within 130 minutes, and react in the dark for 130 minutes after the addition.

[0049] The solution that completed the reaction was put into a 15000 dialysis bag, dialyzed for 48 hours, and the solution in the bag was freeze-dried.

[0050] Through gel chromatographic analysis, th...

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Abstract

The invention relates to a modification method of functional plant polysaccharide, which comprises extracting functional polysaccharide from the plant with biological activity, separating, purifying and determining molecular weight and biological function, modifying structure via agent, activating active hydroxyl on sugar chain, connecting small molecule polyethylene imine, dialyzing, freezing and drying to obtain functional plant polysaccharide. The modification method has simple process, high efficiency and high yield, which keeps prior activity of natural polysaccharide and adds new biological function to obtain a natural macromolecule compound as non-viral gene carrier.

Description

technical field [0001] The invention relates to a class of natural polymer materials, in particular to a method for modifying functional plant polysaccharides. Background technique [0002] Polysaccharides are polymers of aldose and / or ketose sugars that exist in nature linked together by glycosidic bonds. Polysaccharides are essential components of all living organisms, and they are closely related to various physiological functions that sustain life. In recent years, polysaccharides from plants, marine organisms, and fungi have emerged as an important type of biologically active natural products. have been discovered successively. Polysaccharides have a complex structure because they have many different monosaccharide residues, different attachment positions, and different types of glycosidic bonds, which can form different conformations, different relative molecular masses, and intra-chain and inter-chain hydrogen bonds. secondary structure. It is particularly worth n...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K47/36
Inventor 汤谷平周峻姜启英胡秀荣
Owner ZHEJIANG UNIV
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