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CN-121971342-A - Pummelo peel fermentation concentrated solution, preparation method thereof and application thereof in preventing and treating pneumonia and pulmonary fibrosis

CN121971342ACN 121971342 ACN121971342 ACN 121971342ACN-121971342-A

Abstract

The invention discloses a pummelo peel fermentation concentrated solution, a preparation method thereof and application thereof in preventing and treating pneumonia and pulmonary fibrosis. The preparation method comprises the steps of carrying out hot water leaching on exocarpium Citri Grandis, carrying out compound enzymolysis by using cellulase and pectase, inoculating single specific lactobacillus for fermentation, and finally carrying out solid-liquid separation, sterilization and concentration. According to the invention, through an enzymolysis-single bacteria fermentation synergistic process, the naringin content in the product is improved to more than 4000mg/100mL, and the antioxidation activity is obviously enhanced. In vitro pharmacological experiments prove that the fermentation concentrated solution can efficiently relieve the pneumonia induced by LPS and the lung fibrosis induced by TGF-beta by synergistically regulating down key inflammation and fibrosis genes, and the effect is obviously better than that of unfermented and enzymolysis products. The process is green and controllable, and the obtained product has wide application prospect in preparing medicaments for preventing and treating pneumonia and pulmonary fibrosis.

Inventors

  • HUANG JUNYANG
  • CHEN RUIFENG
  • XU HUIHUI
  • XU MEI
  • ZHAO DAZHOU

Assignees

  • 厦门元之道生物科技有限公司

Dates

Publication Date
20260505
Application Date
20260305

Claims (10)

  1. 1. The preparation method of the pummelo peel fermentation concentrated solution is characterized by comprising the following steps: s1, raw material pretreatment, namely crushing pummelo peel, sieving the crushed pummelo peel with a 20-30 mesh sieve, and then mixing the pummelo peel with deionized water according to a mass ratio of 1:20-1:25 to obtain a pummelo peel mixed solution; S2, extracting, namely performing hot water extraction on the pummelo peel mixed solution to obtain pummelo peel water extract; S3, performing enzymolysis, namely adding cellulase with the mass of 0.5% and pectase with the mass of 0.5% into the orange peel water extract, and performing enzymolysis at 50-60 ℃ for 160-180 min to obtain orange peel enzymolysis liquid; s4, sterilizing before fermentation, namely sterilizing and cooling exocarpium Citri Grandis enzymolysis liquid; S5, single strain fermentation, namely inoculating single lactobacillus strain with the liquid system of 0.01% into cooled exocarpium Citri Grandis enzymolysis liquid for fermentation, wherein the fermentation temperature is 36-39 ℃ and the fermentation time is 20 hours, so as to obtain exocarpium Citri Grandis fermentation liquid; The lactobacillus adopts one of lactobacillus plantarum (Lactiplantibacillus plantarum) YYS-99, lactobacillus mucilaginosus (Limosilactobacillus fermentum) JIAN, pediococcus acidilactici (Pediococcus acidilactici) SUN01, lactobacillus paracasei (Lacticaseibacillus paracasei) B20 and lactobacillus plantarum (Lactiplantibacillus plantarum) MPs-68; S6, solid-liquid separation and concentration, namely, carrying out solid-liquid separation on the pummelo peel fermentation liquid, collecting supernatant, sterilizing and concentrating to obtain pummelo peel fermentation concentrated liquid.
  2. 2. The method for preparing pummelo peel fermentation concentrate according to claim 1, wherein the pummelo peel mixed solution is hydrolyzed at 95-100 ℃ for 100-120 min in the step S2.
  3. 3. The method according to claim 1, wherein in step S3, the cellulase activity is 20000 u/g and the pectase activity is 30000 u/g.
  4. 4. The method for preparing pummelo peel fermentation concentrate according to claim 1, wherein the sterilization and cooling conditions in step S4 are that the pummelo peel enzymatic hydrolysate is heated to 95-100 ℃ and kept for 15-20 minutes for sterilization, and cooled to 37+ -2 ℃ after the sterilization is completed.
  5. 5. The method for preparing pummelo peel fermentation concentrate according to claim 1, wherein in step S5, each lactic acid bacteria strain is provided as a bacterial powder; when lactobacillus plantarum YYS-99 is used, the viable count of the bacterial powder is 1-5 multiplied by 1012 CFU/g; When the lactobacillus mucilaginosus JIAN is used, the viable count of the bacterial powder is 1-5 multiplied by 101 0 CFU/g; when Pediococcus acidilactici SUN01 is used, the viable count of the bacterial powder is 1-5 multiplied by 1011 CFU/g; when the lactobacillus paracasei B20 is used, the viable count of the bacterial powder is 1-5 multiplied by 1012 CFU/g; when the lactobacillus plantarum MPs-68 are used, the viable count of the bacterial powder is 1-5 multiplied by 10 9 CFU/g.
  6. 6. The method for preparing pummelo peel fermentation concentrate according to claim 1, wherein in step S6, the pummelo peel fermentation concentrate is first subjected to rough filtration by using a 300-mesh sieve, and then centrifuged at 5000 r/min for 5min, and then the pummelo peel fermentation supernatant is collected; Sterilizing for 20-30 min at 100-110 ℃ to obtain sterilized pummelo peel fermentation liquor; Concentrating sterilized exocarpium Citri Grandis fermentation broth at 70deg.C to 1/7.5 of original volume to obtain exocarpium Citri Grandis fermentation concentrate.
  7. 7. A pummelo peel fermentation concentrate, which is characterized in that the pummelo peel fermentation concentrate is prepared by the preparation method according to any one of claims 1 to 6, and the naringin content in the pummelo peel fermentation concentrate is more than or equal to 4000 mg/100mL.
  8. 8. Use of the pummelo peel fermentation concentrate according to claim 7 in the preparation of food, health care product or cosmetic having an antioxidant function.
  9. 9. Use of the pummelo peel fermentation concentrate according to claim 7 for the preparation of a medicament for the prevention and treatment of pneumonia, wherein the mechanism for the prevention and treatment of pneumonia comprises down-regulating expression of inflammatory factor genes p-65, IRF3, TNF- α and IL-6.
  10. 10. Use of the exocarpium Citri Grandis fermentation concentrate according to claim 7 in the preparation of a medicament for preventing and treating pulmonary fibrosis, wherein the mechanism for preventing and treating pulmonary fibrosis comprises down-regulating expression of fibrosis-related genes Collagen I, vimentin, HDAC, ATG3 and ATG5, and reducing concentration of type I Collagen.

Description

Pummelo peel fermentation concentrated solution, preparation method thereof and application thereof in preventing and treating pneumonia and pulmonary fibrosis Technical Field The invention belongs to the technical field of biotechnology and traditional Chinese medicine deep processing, and particularly relates to a preparation method of pummelo peel fermentation concentrated solution and application of the concentrated solution prepared by the method in preventing and treating pneumonia and pulmonary fibrosis. Background Pneumonia and pulmonary fibrosis are respiratory diseases that severely threaten human health. Pneumonia is mostly caused by excessive inflammatory reaction caused by pathogen infection, and pulmonary fibrosis is mainly characterized by proliferation of pulmonary fibroblasts and massive deposition of extracellular matrix, ultimately resulting in destruction of lung structure and loss of function. At present, the clinic is mainly adopting glucocorticoid and immunosuppressant for treatment, but the drug resistance and various side effects, such as immune function inhibition, osteoporosis and the like, are easy to generate after long-term use. Exocarpium Citri Grandis (CITRI GRANDIS Exocarpium) is a traditional Chinese medicine, has effects of regulating qi-flowing, relieving rigidity of the middle warmer, eliminating dampness and resolving phlegm, and its main active ingredients include flavonoid compounds such as naringin, polysaccharide and volatile oil, etc., and modern pharmacological study shows that it has antiinflammatory, antioxidant and immunoregulatory effects. However, the exocarpium Citri Grandis extract obtained by the traditional water extraction or alcohol extraction method has the problems of low dissolution rate of active ingredients, low bioavailability and the like, and the full play of the drug effect is limited. The microbial fermentation technology is an effective means for improving the drug effect of the traditional Chinese medicinal materials. In the prior art, mixed bacteria are adopted to ferment exocarpium citri grandis, but researches are focused on the general anti-inflammatory and antioxidant effects, and about the prevention and treatment effects and mechanisms of single specific lactic acid bacteria on pneumonia and pulmonary fibrosis, no systematic report is yet available at present. Disclosure of Invention The invention aims to provide a preparation method of pummelo peel fermentation concentrate capable of remarkably improving naringin content and enhancing biological activity. In order to achieve the aim, the solution scheme of the invention is that the preparation method of the pummelo peel fermentation concentrated solution comprises the following steps: s1, raw material pretreatment, namely crushing pummelo peel, sieving the crushed pummelo peel with a 20-30 mesh sieve, and then mixing the pummelo peel with deionized water according to a mass ratio of 1:20-1:25 to obtain a pummelo peel mixed solution; S2, extracting, namely performing hot water extraction on the pummelo peel mixed solution to obtain pummelo peel water extract; S3, performing enzymolysis, namely adding cellulase with the mass of 0.5% and pectase with the mass of 0.5% into the orange peel water extract, and performing enzymolysis at 50-60 ℃ for 160-180 min to obtain orange peel enzymolysis liquid; s4, sterilizing before fermentation, namely sterilizing and cooling exocarpium Citri Grandis enzymolysis liquid; S5, single strain fermentation, namely inoculating single lactobacillus strain with the liquid system of 0.01% into cooled exocarpium Citri Grandis enzymolysis liquid for fermentation, wherein the fermentation temperature is 36-39 ℃ and the fermentation time is 20 hours, so as to obtain exocarpium Citri Grandis fermentation liquid; The lactobacillus adopts one of lactobacillus plantarum (Lactiplantibacillus plantarum) YYS-99, lactobacillus mucilaginosus (Limosilactobacillus fermentum) JIAN, pediococcus acidilactici (Pediococcus acidilactici) SUN01, lactobacillus paracasei (Lacticaseibacillus paracasei) B20 and lactobacillus plantarum (Lactiplantibacillus plantarum) MPs-68; S6, solid-liquid separation and concentration, namely, carrying out solid-liquid separation on the pummelo peel fermentation liquid, collecting supernatant, sterilizing and concentrating to obtain pummelo peel fermentation concentrated liquid. In the step S2, the pummelo peel mixed solution is hydrolyzed for 100-120 min at 95-100 ℃. Further, in the step S3, the enzyme activity of the cellulase is 20000 u/g, and the enzyme activity of the pectase is 30000 u/g. In step S4, the sterilization and cooling conditions are that the exocarpium citri rubrum enzymolysis liquid is heated to 95-100 ℃, and kept for 15-20 minutes for sterilization, and after the sterilization is finished, the exocarpium citri rubrum enzymolysis liquid is cooled to 37+/-2 ℃. Further, in step S5, each lactic acid bacteria strain is provided as a ba