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CN-118240109-B - Grifola frondosa polysaccharide GFP22, preparation method thereof and application thereof in preparation of prebiotic products and auxiliary antibacterial drugs

CN118240109BCN 118240109 BCN118240109 BCN 118240109BCN-118240109-B

Abstract

The invention discloses a grifola frondosa polysaccharide GFP22, a preparation method thereof and application thereof in preparing prebiotic products and auxiliary antibacterial medicines. The grifola frondosa polysaccharide GFP22 is mannose and glucose polymerized to form mannose and glucose, the ratio of mannose to glucose is 2.88:5.24, the grifola frondosa polysaccharide GFP has the following structure, and the weight average molecular weight range is 4.9kDa. The grifola frondosa polysaccharide GFP22 can promote the synthesis of short-chain fatty acid in intestinal tracts, regulate the steady state of intestinal flora, promote the proliferation of bifidobacteria and inhibit the proliferation of klebsiella, and is hopeful to be developed into a prebiotic product and a saccharide medicament for assisting in resisting the klebsiella. R:β-D-Glcp-(1→6)-β-D-Manp-(1→。

Inventors

  • CHEN SHAODAN
  • CAI MANJUN
  • LIU YUANCHAO
  • WU QINGPING
  • HU HUIPING
  • XIE YIZHEN
  • BAI GUANGJIAN
  • GAO XIONG
  • XIAO CHUN
  • YONG TIANQIAO
  • HUANG LONGHUA

Assignees

  • 广东省科学院微生物研究所(广东省微生物分析检测中心)

Dates

Publication Date
20260505
Application Date
20240409

Claims (10)

  1. 1. The preparation method of the grifola frondosa polysaccharide GFP22 is characterized by comprising the following steps: (1) Polysaccharide extraction, namely extracting the fruiting body of the grifola frondosa by boiling water, collecting the extracting solution, concentrating and centrifuging, and then dialyzing and precipitating the centrifugate supernatant, centrifuging by alcohol precipitation, and collecting precipitate to obtain crude polysaccharide GFP; (2) And (3) polysaccharide purification, namely separating crude polysaccharide GFP by a DEAE Sepharose TM Fast Flow anion exchange column, sequentially using deionized water, 0.1M and 0.2M NaCl solution for gradient elution, collecting an eluting peak component eluted by the 0.1M NaCl solution to obtain secondary crude polysaccharide GFP2, further purifying the secondary crude polysaccharide GFP2 by a SEPHACRYL S-200HR gel column, eluting by using the NaCl solution with the pH of 6.8 and the concentration of 0.05M phosphate buffer salt of 0.15M, and collecting an eluting component to obtain the grifola frondosa polysaccharide GFP22.
  2. 2. The method according to claim 1, wherein in the step (1), ethanol used for precipitation is 95% ethanol by volume, and the volume of ethanol used for precipitation is 3 to 5 times that of the concentrated solution.
  3. 3. The Grifola frondosa polysaccharide GFP22 prepared by the preparation method of claim 1.
  4. 4. The grifola frondosa polysaccharide GFP22 according to claim 3, wherein the grifola frondosa polysaccharide GFP22 is a mannans polymerized by mannose and glucose, and has the following structure: R: β-D-Glcp-(1→6)-β-D-Manp-(1→。
  5. 5. The ash tree flower polysaccharide GFP22 according to claim 4, wherein the ash tree flower polysaccharide GFP22 has a weight average molecular weight of 4.90 kDa, and a ratio of mannose to glucose of 2.88:5.24.
  6. 6. Use of the grifola frondosa polysaccharide GFP22 of any one of claims 3-5 in the preparation of a prebiotic product or an auxiliary antibacterial medicament.
  7. 7. The use according to claim 6, wherein the prebiotic product promotes the production of short chain fatty acids in the gut and regulates gut flora homeostasis.
  8. 8. The use according to claim 7, wherein the short chain fatty acids comprise acetic acid, propionic acid and butyric acid, and wherein the modulation of intestinal flora homeostasis comprises promotion of bifidobacterium proliferation and inhibition of klebsiella proliferation.
  9. 9. The use according to claim 6, wherein the antibacterial object of the auxiliary antibacterial agent comprises klebsiella pneumoniae.
  10. 10. A prebiotic product or auxiliary antibacterial agent comprising as active ingredient the grifola frondosa polysaccharide GFP22 of any one of claims 3-5.

Description

Grifola frondosa polysaccharide GFP22, preparation method thereof and application thereof in preparation of prebiotic products and auxiliary antibacterial drugs Technical Field The invention belongs to the field of biological medicine, and in particular relates to a grifola frondosa polysaccharide GFP22, a preparation method thereof and application thereof in preparation of prebiotic products and auxiliary antibacterial medicines. Background Intestinal flora refers to microorganisms in the intestinal tract, and about 10 trillion microorganisms exist in the intestinal tract of a human, and the microorganisms play a very important role in regulating nutrient absorption, maintaining epithelial cell development, regulating innate immunity and the like. The homeostasis and diversity of intestinal flora are closely related to host cell proliferation, neural signaling pathways, etc. Unbalanced intestinal flora, excessive proliferation of harmful bacteria, reduced beneficial bacteria, and can cause various diseases such as hypoimmunity and metabolic disorder. Klebsiella (Klebsiella Pneumoniae, KP) belongs to the Enterobacteriaceae family, and is a gram-negative facultative anaerobe, commonly found in the environment, such as water, soil and vegetation, and can colonize the respiratory tract or intestinal tract of humans or animals, and is a common zoonotic conditional pathogen. Klebsiella pneumoniae (Klebsiella Pneumoniae, KP) is one of the most popular opportunistic pathogens in the world at present, has high pathogenicity to human beings, animals and the like, and can cause pneumonia, metritis, mastitis and other suppurative inflammations of human beings and animals, even septicemia. Edible fungi polysaccharide is carbohydrate polymer derived from edible fungi, is resistant to endogenous digestive enzymes, and is neither hydrolyzed nor absorbed in small intestine. Since edible fungus polysaccharide cannot be digested and degraded by human body, and the intestinal flora planted in the intestinal lumen of human body contains abundant polysaccharide hydrolase, the edible fungus polysaccharide is mainly metabolized and degraded by the intestinal flora and affects the intestinal flora, so that the health of a host is regulated. Such healthy polysaccharides or dietary fibers are also commonly referred to as prebiotics, which promote the proliferation of the beneficial bacteria corresponding thereto, and which upon administration in certain amounts provide a host health benefit to the microorganism. Grifola frondosa (Grifola frondosa) is a fungus of Grifola genus (Grifola) of Polyporaceae family (Polyporaceae), and is one of rare edible and medicinal fungi developed and utilized in recent years. The Maitake Mushroom contains chemical components such as polysaccharide, sterol and polyphenols, and has effects of resisting tumor, regulating immunity, reducing blood sugar and blood lipid, etc. Grifola frondosa strain HMGIM-W151021 is a high-yield polysaccharide strain which is collected from the wild and successfully domesticated by the university of Guangdong microbiological institute, and the preservation number of the strain is GDMCC No:61165, which is disclosed in Chinese patent ZL 202210673582.4, however, the polysaccharide structure and the regulation and control effect of the polysaccharide structure on intestinal flora are not reported yet. Disclosure of Invention The invention uses a simple and effective process and method for extracting and purifying edible fungus polysaccharide, takes the Grifola frondosa HMGIM-W151021 as a raw material to obtain a mixed polysaccharide extract, and further carries out purification to obtain the Grifola frondosa polysaccharide GFP22 with uniform charge and molecular weight. In vitro experiments prove that the polysaccharide can regulate the balance of intestinal flora, inhibit pathogenic bacteria of intestinal klebsiella, promote probiotics such as bifidobacteria and the like. And after the polysaccharide is subjected to intestinal anaerobic fermentation, short-chain fatty acid can be produced, and the short-chain fatty acid can regulate the immunity and metabolic homeostasis of organisms, so that the polysaccharide is hopeful to be developed into a novel prebiotic or a saccharide medicament for assisting in resisting pneumonia and related diseases caused by klebsiella. The invention provides a grifola frondosa polysaccharide GFP22, which is mannose-glucose with a weight average molecular weight of 4.90kDa, and has the following structure: R:β-D-Glcp-(1→6)-β-D-Manp-(1→。 the ash tree flower polysaccharide GFP22 is polysaccharide polymerized by mannose and glucose, and the ratio (molar ratio) of the mannose to the glucose is 2.88:5.24. The invention also provides a preparation method of the grifola frondosa polysaccharide GFP22, which comprises the following steps: (1) Polysaccharide extraction, namely extracting the fruiting body of the grifola frondosa by boiling water, collecting