CN-121991259-A - High-activity rhizoma polygonati polysaccharide and preparation method and application thereof

Abstract

The invention provides a high-activity rhizoma polygonati polysaccharide, a preparation method and application thereof, and belongs to the technical field of active ingredient extraction. The invention takes the mixture of cellulase, hemicellulase and pectase as a complex enzyme preparation to carry out enzymolysis, extraction and purification to obtain the rhizoma polygonati polysaccharide PCPA. The molecular weight of the rhizoma polygonati polysaccharide PCPA is 1372 Da, the rhizoma polygonati polysaccharide consists of three monosaccharides of arabinose, glucose and fructose, the molar ratio of the three monosaccharides is 0.39:6.48:93.13, and the PCPA has the effects of delaying cell aging and improving cell repair activity.

Inventors

• YAN SHUPING
• WANG JIANCHENG
• SONG SHUANGSHUANG
• ZHANG HUIMIN
• ZHANG LIMEI
• XING YAN

Assignees

• 国珍健康科技（北京）有限公司

Dates

Publication Date

20260508

Application Date

20260317

Claims (10)

1. 1. A preparation method of high-activity rhizoma polygonati polysaccharide is characterized by comprising the following steps: (1) Pulverizing rhizoma Polygonati, and sieving with 20-40 mesh sieve to obtain rhizoma Polygonati powder; (2) Mixing rhizoma Polygonati powder with distilled water, regulating pH, and adding compound enzyme preparation at 40-60deg.C for enzymolysis at 2-4 h to obtain enzymolysis solution; (3) Extracting by heating and extracting enzymolysis solution at 100deg.C for 1-2 h, centrifuging at 5000: 5000 r/min, collecting supernatant, concentrating under reduced pressure to 1/5 of the original volume to obtain concentrated solution; (4) Precipitating with ethanol to remove impurities, adding absolute ethanol to reach ethanol concentration of 80%, standing at 4deg.C for 12 h, collecting precipitate, and lyophilizing to obtain rhizoma Polygonati crude polysaccharide; (5) Purifying by gel chromatography column, and lyophilizing to obtain single homogeneous rhizoma Polygonati active polysaccharide PCPA.
2. 2. The method of claim 1, wherein the mass/volume ratio of the rhizoma polygonati powder to distilled water in the step (2) is 1 g/15-25 mL.
3. 3. The process according to claim 1, wherein the pH in step (2) is 6.
4. 4. The method of claim 1, wherein the mass ratio of the cellulase, the hemicellulase and the pectinase is 3:1-3:1-3.
5. 5. The method of claim 1, wherein the amount of the complex enzyme preparation is 0.5-2% of the mass of the rhizoma polygonati powder.
6. 6. The method according to claim 1, wherein the gel column in the step (5) is a sephadex G-50.
7. 7. The preparation method of the polygonatum sibiricum polysaccharide powder, as set forth in claim 1, is characterized in that the purification specific steps in the step (5) are that polygonatum sibiricum polysaccharide is taken and dissolved into 40-60mg/mL solution, the solution is filtered through a 0.45 mu m filter membrane for later use, a chromatographic column is packed by a wet method, the diameter-to-height ratio is 1:15, pure water is used for elution, the flow rate is set to be 2 mL/min,3 min/tube is used for collecting eluent, the content of polysaccharide in each collected liquid is detected by a phenol-sulfuric acid method, a crude polysaccharide elution curve of polygonatum sibiricum is drawn, a single peak is collected, and freeze drying is carried out, so that single homogeneous polygonatum sibiricum active polysaccharide PCPA is obtained.
8. 8. A high-activity Polygonatum sibiricum polysaccharide prepared by the preparation method of any one of claims 1-7 is characterized by comprising three monosaccharides of arabinose, glucose and fructose in a molar ratio of 0.39:6.48:93.13, wherein the high-activity Polygonatum sibiricum polysaccharide has Mn molecular weight of 1372 Da, MP molecular weight of 2402 Da and Mw molecular weight of 2372 Da.
9. 9. The use of the high-activity rhizoma Polygonati polysaccharide according to any one of claims 1-7 in preparing products with antiaging effect.
10. 10. The method according to claim 9, wherein the product is any one or more of the following: (1) The product is a product capable of improving the cell repair activity; (2) The product can reduce the SA-beta-gal content in cells; (3) The product is a product capable of increasing the intracellular NAD+ content.

Description

High-activity rhizoma polygonati polysaccharide and preparation method and application thereof Technical Field The invention belongs to the technical field of extraction of active ingredients, and particularly relates to a high-activity polygonatum polysaccharide, and a preparation method and application thereof. Background Rhizoma Polygonati is used as a traditional medicine and food homologous resource, and is originally in Ming Yi Bie Lu. Rhizoma Polygonati has effects of delaying aging, enhancing immunity, reducing blood sugar, reducing blood lipid, relieving inflammation, resisting bacteria and virus, and resisting tumor. Rhizoma Polygonati contains polysaccharide, saponin, anthraquinone compounds, alkaloid, cardiac glycoside, lignin, vitamins and other active components beneficial to human body, and its core active component, rhizoma Polygonati polysaccharide (Polygonatum Cyrtonema Polysaccharides, PCPs) has been attracting attention because of its wide bioactivity. Under the market driving of pursuing natural and efficient active ingredients, developing rhizoma polygonati polysaccharide products with definite targeting effects (such as anti-aging and skin repair) has become a research hotspot. At present, the extraction method of the rhizoma polygonati polysaccharide mainly comprises a hot water extraction method, an acid-base extraction method, an enzyme-assisted extraction method and the like. Although the hot water extraction method is simple and convenient to operate, the extraction temperature is high, the extraction time is long, the polysaccharide yield is low, the polysaccharide structure is easy to damage, and the acid-base extraction method can cause the polysaccharide glycosidic bond to break, so that the biological activity of the polysaccharide is influenced. The enzyme-assisted extraction technology is gradually applied to the extraction research of plant polysaccharide due to mild acting conditions and high extraction efficiency. However, the prior enzymolysis technology mostly adopts a single enzyme preparation, has limited cell wall degradation capability, is difficult to fully release the rhizoma polygonati intracellular polysaccharide, and meanwhile, the crude rhizoma polygonati polysaccharide component obtained by conventional extraction is complex, has wide molecular weight distribution range and poor uniformity, and the active polysaccharide component truly playing a key role is masked by a large number of invalid or low-efficiency components, so that the overall biological activity is not outstanding and is unstable. And the high molecular weight polysaccharide has certain activity, but has poor water solubility, diffusion capability in a biological system and transdermal absorption rate, and is difficult to effectively penetrate the skin stratum corneum to reach target cells in skin care application. In addition, the heterogeneous mixture does not allow the establishment of an accurate "structure-activity" relationship, severely hampering standardization and efficacy claims of the product. The related patent documents: The preparation method comprises (1) dissolving Polygonatum cyrtonema Fabricius polysaccharide in buffer solution of enzymolysis reaction, (2) adding fructosidase for enzymolysis reaction, and (3) centrifuging, dialyzing, purifying, and lyophilizing the reaction solution sequentially after terminating the reaction. According to the invention, the degradation fragments of the polygonatum cyrtonema polysaccharide are prepared by an enzymatic degradation method, the activity of promoting GLP-1 secretion is used as a guide, various parameters of enzymolysis reaction are optimized, and the oligosaccharide fragments with high activity in the polygonatum cyrtonema polysaccharide are obtained by screening, so that the method has the advantages of simplicity in operation, strong specificity, environment friendliness and the like, and provides scientific basis and theoretical basis for development of natural GLP-1 agonist hypoglycemic drugs, but the preparation method is complicated and single homogeneous polysaccharide cannot be obtained. A method for preparing neutral oligomeric polysaccharide from rhizoma Polygonati is also disclosed in Chinese patent CN 112480283A. The method comprises the steps of (1) leaching and solid-liquid separating rhizoma polygonati raw materials to obtain leaching liquor, concentrating, freezing, melting and solid-liquid separating the leaching liquor, collecting clear liquid, (2) subjecting the clear liquid to enzyme column chromatography, sequentially passing through a large-aperture membrane and a small-aperture membrane, respectively collecting trapped liquid, and (3) subjecting the trapped liquid of the small-aperture membrane to ion exchange resin, then adjusting pH to 6-7, concentrating, alcohol precipitating and drying. The method has the advantages of mild process conditions, strong continuity, low requirements on equipment, strong o