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CN-121971480-A - Fresh rehmannia root cell wall polysaccharide composition, extraction method and application

CN121971480ACN 121971480 ACN121971480 ACN 121971480ACN-121971480-A

Abstract

The invention relates to a fresh rehmannia root cell wall polysaccharide composition, an extraction method and application thereof, belonging to the field of plant extraction and application, the invention adopts a sequential extraction method to gradually extract cell wall polysaccharide from FRR, and three cell wall pectic polysaccharides (WSP, CSP, NSP) and two hemicellulose polysaccharides (KSH-1 and KSH-2) are successfully obtained. The results show that CSP and NSP exhibit relatively uniform molecular weight distribution. In terms of composition, WSP, CSP and NSP are composed mainly of Ara and GalA, while KSH-1 and KSH-2 contain mainly Ara and Xyl. The FRR-derived polysaccharide has been shown to have good prospects as a functional food or pharmaceutical ingredient having antioxidant and immunoregulatory functions.

Inventors

  • WANG LI
  • MA BINGJI
  • PENG HUASHENG
  • YANG YUPENG
  • WEN CHUNNAN
  • SONG XIAOLONG

Assignees

  • 河南农业大学

Dates

Publication Date
20260505
Application Date
20260213

Claims (10)

  1. 1. A fresh rehmannia cell wall polysaccharide composition is characterized by comprising water-soluble pectin, CDTA-soluble pectin, na 2 CO 3 -soluble pectin, hemicellulose KSH1 and KSH-2, wherein the water-soluble pectin, CDTA-soluble pectin and NA 2 CO 3 -soluble pectin comprise main components of arabinose, galactose and galacturonic acid, monosaccharides such as Fuc, man and Xyl are not contained, the ratio of arabinose in the water-soluble pectin, CDTA-soluble pectin and NA 2 CO 3 -soluble pectin is 28.72%, 38.46% and 42.36%, the ratio of galacturonic acid is 46.26%, 25.43% and 20.40%, the ratio of galactose is 12.02%, 15.32% and 20.73%, the ratio of arabinose in the KSH-1 and KSH-2 is 55.53% and 30.03%, and the ratio of galactose is 13.53% and 11.62% and 35.22% respectively; The total content of galacturonic acid and glucuronic acid in the water-soluble pectin, CDTA-soluble pectin, NA 2 CO 3 -soluble pectin is 53.18%, 38.51% and 34.52%, HG=GalA-Rha, RG-1=2×Rha+Glc+Ara, HG ratio of the water-soluble pectin, CDTA-soluble pectin and NA 2 CO 3 -soluble pectin is 43.94%, 21.94% and 15.23%, RG-I ratio is 35.71%, 45.44% and 52.70%, respectively, and the ratio of Rha/(GalA+GlcA) in the water-soluble pectin, CDTA-soluble pectin and NA 2 CO 3 -soluble pectin is 0.04, 0.09 and 0.15, respectively, the Xyl/Ara ratio of KSH-1 is 0.2, and the Xyl/Ara ratio of KSH-2 is 1.17.
  2. 2. The cell wall polysaccharide composition of claim 1, wherein the cell wall polysaccharide composition comprises: The water-soluble pectin has two peaks with molecular weights of 8.638 X10 5 g/mol and 1.479X 10 5 g/mol, respectively; the CDTA soluble pectin has a single peak and a molecular weight of 1.893 X10- 5 g/mol; The NA 2 CO 3 soluble pectin has a single peak and a molecular weight of 2.149 multiplied by 10 5 g/mol; The molecular weight of the KSH-1 is 1.773 multiplied by 10 4 g/mol; The molecular weight of the KSH-2 is 9.195 X10- 4 g/mol.
  3. 3. The composition of claim 1, wherein the water-soluble pectin has a degree of esterification of 71.52%, the CDTA-soluble pectin has a degree of esterification of 53.63%, and the NA 2 CO 3 -soluble pectin has a degree of esterification of 72.04%.
  4. 4. The composition of claim 1, wherein the water-soluble pectin, CDTA-soluble pectin and NA 2 CO 3 -soluble pectin each have a triple helix structure.
  5. 5. The cell wall polysaccharide composition of claim 1, wherein the cell wall polysaccharide composition has pseudoplastic fluid behavior, has apparent viscosity in the range of 0.01-100 s -1 , increases with increasing concentration, and is significantly enhanced by Ca 2+ ions.
  6. 6. A method for extracting a cell wall polysaccharide composition of fresh rehmannia root as defined in any one of claims 1 to 5, comprising the steps of: (1) Preparing fresh rehmannia alcohol insoluble matter, namely cleaning fresh rehmannia root and stem, cutting, refluxing and boiling with 95% ethanol, degreasing residues with chloroform/methanol after filtration, removing starch with DMSO, and finally drying to obtain fresh rehmannia alcohol insoluble matter; (2) Sequentially extracting cell wall polysaccharide, namely sequentially treating AIR with the following solvents: a. Extracting with water at 25deg.C for 16 hr to obtain water-soluble pectin; b. CDTA solution (0.05M, pH 6), extracted at 25 ℃ for 18 hours to obtain CDTA soluble pectin; c. NA 2 CO 3 solution (0.05M, containing 0.02, M NaBH 4 ) was extracted at 4 ℃ for 20 hours and then at 25 ℃ for 2 more hours to obtain NA 2 CO 3 soluble pectin; d. 1M KOH (containing 0.02M NaBH 4 ), extracting at 25deg.C under nitrogen for 18 hr to obtain hemicellulose KSH-1; e. 4M KOH (containing 0.02M NaBH 4 ), extracting at 25deg.C under nitrogen for 2 hr to obtain hemicellulose KSH-2; (3) Purifying by concentrating the extractive solutions, dialyzing (molecular weight cut-off 3500 Da), and lyophilizing to obtain five cell wall polysaccharide components.
  7. 7. Use of the prepared cell wall polysaccharide composition of rehmannia root as defined in claim 6 for preparing products for promoting proliferation and migration of RAW264.7 cells and regulating secretion of cytokines, including inhibiting release of IL-10 and promoting secretion of NO, TNF- α and IL-6.
  8. 8. The method according to claim 7, wherein the immunomodulatory activity is positively correlated with galacturonic acid content and negatively correlated with molecular weight.
  9. 9. Use of the cell wall polysaccharide composition of rehmannia glutinosa ('suk') in the preparation of an antioxidant.
  10. 10. The method according to claim 9, wherein the antioxidant activity is positively correlated with the molecular weight and arabinose content of the polysaccharide, and wherein the correlation coefficient r=0.86 between the radical scavenging capacity of ABTS + and the molecular weight.

Description

Fresh rehmannia root cell wall polysaccharide composition, extraction method and application Technical Field The invention relates to a fresh rehmannia root cell wall polysaccharide composition, an extraction method and application thereof, and belongs to the field of plant extraction and application. Background Fresh rehmannia (FRR) is a commonly used traditional Chinese medicinal material. According to the description of Ben Cao gang mu, it has the functions of clearing heat, cooling blood, stopping bleeding and detoxicating. Polysaccharide is considered as one of the main active ingredients of rehmannia glutinosa. In recent years, researches show that rehmannia polysaccharide has the functions of enhancing immunity, resisting oxidation, reducing blood sugar, resisting aging and the like. In the prior art, polysaccharide in dried rehmannia root and prepared rehmannia root is separated, extracted and activity studied, such as CN202111303342.7, CN202311589402.5, CN202211362527.X, and the like, mainly prepared rehmannia root, dried rehmannia root decoction pieces or fresh rehmannia root leaves/stems are used as raw materials to extract rehmannia root polysaccharide, so that the physicochemical properties of the rehmannia root polysaccharide are studied. However, there are few reports on the research on the cell wall polysaccharide of the fresh rehmannia, so that the comprehensive and systematic research on the cell wall polysaccharide (FRR CWPs) of the fresh rehmannia has important practical significance. Disclosure of Invention The invention provides a fresh rehmannia root cell wall polysaccharide composition, an extraction method and application thereof, solves the problem that the research on the fresh rehmannia root cell wall polysaccharide is not carried out in the existing rehmannia root polysaccharide research, and effectively expands the development and application of the fresh rehmannia root cell wall polysaccharide. In order to solve the technical problems, the technical scheme adopted by the invention is that the fresh rehmannia root cell wall polysaccharide composition is characterized by comprising water-soluble pectin, CDTA-soluble pectin, NA 2CO3 -soluble pectin, hemicellulose KSH1 and KSH-2, wherein the main components of the water-soluble pectin, the CDTA-soluble pectin and the NA 2CO3 -soluble pectin are arabinose, galactose and galacturonic acid, the proportion of the arabinose in the water-soluble pectin, the CDTA-soluble pectin and the NA 2CO3 -soluble pectin is 28.72 percent, 38.46 percent and 42.36 percent respectively, the proportion of the galacturonic acid is 46.26 percent, 25.43 percent and 20.40 percent respectively, the proportion of the galactose is 12.02 percent, 15.32 percent and 20.73 percent respectively, the proportion of the arabinose in the KSH-1 and the KSH-2 is 55.53 percent and 30.03 percent respectively, and the proportion of the xylose in the KSH-1 and the KSH-2 is 13.53 percent and the proportion of the galactose is 11.62 percent and the proportion of the xylose is 35.22.93 percent respectively; The total content of galacturonic acid and glucuronic acid in the water-soluble pectin, CDTA-soluble pectin, NA 2CO3 -soluble pectin is 53.18%, 38.51% and 34.52%, HG=GalA-Rha, RG-1=2×Rha+Glc+Ara, HG ratio of the water-soluble pectin, CDTA-soluble pectin and NA 2CO3 -soluble pectin is 43.94%, 21.94% and 15.23%, RG-I ratio is 35.71%, 45.44% and 52.70%, respectively, and the ratio of Rha/(GalA+GlcA) in the water-soluble pectin, CDTA-soluble pectin and NA 2CO3 -soluble pectin is 0.04, 0.09 and 0.15, respectively, the Xyl/Ara ratio of KSH-1 is 0.2, and the Xyl/Ara ratio of KSH-2 is 1.17. Further, it is preferable that the water-soluble pectin has two peaks, and has a molecular weight of 8.638 X10 5 g/mol and 1.479X 10 5 g/mol, respectively; the CDTA soluble pectin has a single peak and a molecular weight of 1.893 X10- 5 g/mol; The NA 2CO3 soluble pectin has a single peak and a molecular weight of 2.149 multiplied by 10 5 g/mol; The molecular weight of the KSH-1 is 1.773 multiplied by 10 4 g/mol; The molecular weight of the KSH-2 is 9.195 X10- 4 g/mol. Further, preferably: The esterification degree of the water-soluble pectin is 71.52%, the esterification degree of the CDTA-soluble pectin is 53.63%, and the esterification degree of the NA 2CO3 -soluble pectin is 72.04%. Further, preferably, the water-soluble pectin, CDTA-soluble pectin and NA 2CO3 -soluble pectin all have triple helix structures. Further, preferably, the fresh rehmannia root cell wall polysaccharide composition has pseudoplastic fluid behavior, has apparent viscosity in the range of 0.01-100 s -1 of shear rate, increases with increasing concentration, and is significantly enhanced by Ca 2+ ions. The extraction method of the fresh rehmannia root cell wall polysaccharide composition comprises the following steps: (1) Preparation of fresh rehmannia alcohol insoluble substance (AIR) comprises cleaning fresh rehmannia