Medicinal and edible leavening with uric acid reducing effect and preparation method of medicinal and edible leavening

Abstract

The invention belongs to a traditional Chinese medicine fermentation technology, and particularly relates to a medicinal and edible leavening with a uric acid reducing effect. The fermented product is prepared from the following components in parts by weight: 80 to 120 parts of alfalfa, 35 to 65 parts of folium cortex eucommiae, 30 to 60 parts of sparassis crispa and 10 to 20 parts of lucid ganoderma. The components have the effect of reducing uric acid after being subjected to enzymolysis and fermentation, and a medicinal and edible leavening is provided and can be applied to products for relieving hyperuricemia, gout and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a medicine and food homologous fermented product with the efficacy of reducing uric acid and a preparation method thereof. Background Art

[0002] Uric acid is the end metabolite of purine metabolism in the body. When purine metabolism in the body is abnormal or uric acid excretion is blocked, the concentration of blood uric acid in the body increases, leading to hyperuricemia (HUA). The increase in serum uric acid concentration causes monosodium urate crystals to deposit in and around the joint cavity, resulting in the onset of gout. The development of gout will further damage the kidneys. Hyperuricemia is the fourth major underlying disease after hypertension, diabetes, and hyperlipidemia, and has become a health problem that cannot be ignored. Currently, the main drugs used to treat HUA and gout are allopurinol, colchicine, indomethacin, probenecid, benzbromarone, and glucocorticoids, etc. These drugs have the effects of anti-inflammatory, analgesic, inhibiting uric acid production, and promoting uric acid excretion. However, long-term use can cause digestive tract damage, kidney and liver toxicity. And most drug treatments are accompanied by adverse reactions, such as headache, nausea, diarrhea, rash, gastrointestinal reactions, as well as elevated transaminases, decreased white blood cells, urinary tract stones, and reduced renal function.

[0003] Chinese herbal medicines have the advantages of safety, small toxic and side effects, stable curative effect, and economy in reducing uric acid. Chinese herbal medicines for both medicine and food can be used as drugs and are also the main foods in people's daily lives, having great potential in the later product development.

[0004] Modern traditional Chinese medicine fermentation technology, as an emerging technology, can enhance the active ingredients of Chinese herbal medicines, improve bioavailability, or convert complex Chinese herbal medicine components into forms that are more easily absorbed by the human body through the fermentation of microorganisms, thereby improving the medicinal efficacy. Summary of the Invention

[0005] Therefore, the present invention provides a medicine and food homologous fermented product with the efficacy of reducing uric acid and a preparation method thereof. The medicine and food homologous fermented product includes four raw materials, namely alfalfa, eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum, and is prepared by fermentation with probiotics, having the efficacy of reducing uric acid.

[0006] The technical content of the present invention is as follows:

[0007] On the one hand, the present invention provides a medicine and food homologous fermented product with the efficacy of reducing uric acid. The raw materials, by weight, include the following components: 80 - 120 parts of alfalfa, 35 - 65 parts of eucommia ulmoides leaves, 30 - 60 parts of Sparassis crispa, and 10 - 20 parts of Ganoderma lucidum.

[0008] Preferably, the fermented product, in terms of parts by weight of raw materials, comprises the following components: 90-110 parts of alfalfa, 40-60 parts of eucommia ulmoides leaves, 35-55 parts of sparassis crispa, and 10-15 parts of ganoderma lucidum.

[0009] Preferably, it further comprises 0.15-0.27 parts of fermenting agent and 12-22 parts of enzyme preparation.

[0010] More preferably, the fermenting agent is 0.17-0.24 parts, and the enzyme preparation is 14-20 parts.

[0011] More preferably, the fermented product, in terms of parts by weight of raw materials, comprises the following components: 100 parts of alfalfa, 50 parts of eucommia ulmoides leaves, 50 parts of sparassis crispa, 15 parts of ganoderma lucidum, 0.215 parts of fermenting agent, and 17.2 parts of enzyme preparation.

[0012] Preferably, the medicated and edible fermented product further comprises 5-10 parts of astragalus membranaceus.

[0013] More preferably, the medicated and edible fermented product further comprises 8-10 parts of astragalus membranaceus.

[0014] Preferably, the fermenting agent comprises lactobacillus rhamnosus and bifidobacterium animalis subsp. lactis.

[0015] More preferably, the fermenting agent comprises 0.08-0.15 parts of lactobacillus rhamnosus and 0.07-0.12 parts of bifidobacterium animalis subsp. lactis.

[0016] More preferably, the weight ratio of lactobacillus rhamnosus to bifidobacterium animalis subsp. lactis in the fermenting agent is 3:2.

[0017] Even more preferably, the viable count of lactobacillus rhamnosus is 100-5000 billion CFU / g, and the viable count of bifidobacterium animalis subsp. lactis is 100-5000 billion CFU / g.

[0018] Preferably, the enzyme preparation comprises cellulase, pectinase, and papain.

[0019] More preferably, the enzyme preparation comprises 8-12 parts of cellulase, 2-6 parts of pectinase, and 2-4 parts of papain.

[0020] Even more preferably, the weight ratio of cellulase, pectinase, and papain is 3:1:1.

[0021] Even more preferably, the enzyme activity of cellulase is 10,000-100,000 U / g, the enzyme activity of pectinase is 10,000-50,000 U / g, and the enzyme activity of papain is 20,000-150,000 U / g.

[0022] On the other hand, the present invention also provides a preparation method of a fermented product of medicated and edible homologous substances with the effect of reducing uric acid, comprising the following steps:

[0023] (1) Mix the raw materials with water to obtain a mixed solution;

[0024] (2) Adjust the pH of the mixed solution to 4-5, then add an enzyme preparation and perform enzymatic hydrolysis treatment to obtain an enzymatic hydrolysate;

[0025] (3) Adjust the pH of the enzymatic hydrolysate to 5.5-6.5, raise the temperature, and perform enzyme inactivation extraction treatment to obtain an enzyme-inactivated extraction solution;

[0026] (4) Add a fermenting agent to the enzyme-inactivated extraction solution for fermentation to obtain a fermentation broth;

[0027] (5) Centrifuge the fermentation broth, and the supernatant is the fermentation clear solution.

[0028] Preferably, it further comprises the steps of subjecting the fermentation clear solution to reduced pressure concentration and vacuum drying at 60-80 °C to obtain a fermented product.

[0029] Preferably, in step (2), the enzymatic hydrolysis temperature is 40-60 °C and the time is 40-90 min.

[0030] Preferably, in step (3), the enzyme inactivation extraction temperature is 110-121 °C and the enzyme inactivation extraction time is 35-70 min.

[0031] Preferably, the fermentation temperature in step (4) is 35-42 °C and the fermentation time is 18-48 h.

[0032] On the other hand, the present invention provides the application of the above uric acid-lowering composition in the preparation of products for treating hyperuricemia and / or gout.

[0033] Compared with the prior art, the present invention has the following advantages:

[0034] 1. The fermented product of medicated and edible homologous substances provided by the present invention selects a raw material composition with a history of dual use as medicine and food, and has the characteristics of natural safety, low toxicity, and low irritation, and is suitable for long-term use. Alfalfa contains various bioactive ingredients such as saponins, flavonoid compounds, polysaccharides, and coumarins, and can be used for the prevention and treatment of arthritis, gout, hepatitis, cholecystitis, kidney stones and other diseases; Eucommia ulmoides leaves are the dried leaves of Eucommia ulmoides Oliv. of the Eucommiaceae family, and have the effects of tonifying the liver and kidney, astringing essence and arresting seminal emission; Sparassis crispa is a rare edible and medicinal fungus, rich in polysaccharides, proteins, cellulose, minerals and various bioactive ingredients, and has anti-inflammatory, immunomodulatory and other effects; Ganoderma lucidum contains active ingredients such as polysaccharides and phenols, and has the effects of replenishing qi and calming the mind, relieving cough and asthma. The uric acid-lowering effect produced by the synergistic action of the four raw materials is superior to the uric acid-lowering effect of each raw material.

[0035] 2. The medicated and edible ferment with the effect of reducing uric acid provided by the present invention may also include astragalus root. After adding astragalus root, the uric acid-reducing level of the fermentation product can be improved. This may be because astragalus root contains flavonoids (such as calycosin-7-O-β-D-glucoside, calycosin, ononin, formononetin, etc.) and saponins (such as astragaloside IV, astragalosides, etc.). During the co-enzymatic hydrolysis and fermentation process of astragalus root and other raw materials, the flavonoids and saponins can be released to the greatest extent, thus achieving an increase in the uric acid-reducing level.

[0036] 3. The medicated and edible ferment provided by the present invention uses the enzymatic hydrolysis-fermentation technology process to make full use of raw materials, convert macromolecular substances that are difficult to absorb into small molecular substances that are easy to absorb, and improve the bioavailability. Detailed implementation manners

[0037] The following non-limiting examples can enable those of ordinary skill in the art to understand the present invention more comprehensively, but do not limit the present invention in any way. The following content is only an exemplary illustration of the scope claimed by the present invention. Those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and it should also fall within the scope claimed by the present invention.

[0038] When the examples give numerical ranges, it should be understood that unless otherwise specified in the present invention, both endpoints of each numerical range and any value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present invention belongs.

[0039] The present invention will be further described below by way of specific examples. All kinds of chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Products from different manufacturers do not have a significant impact on the effect.

[0040] Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis were purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd., Lactobacillus rhamnosus JYLR-005 and Bifidobacterium animalis subsp. lactis JYBR-190; cellulase and pectinase were purchased from Shandong Longkete Enzyme Preparation Co., Ltd.; papain was purchased from Shaanxi Chenming Technology Co., Ltd.

[0041] Example 1

[0042] This example provides a fermented product of medicine and food homology with the effect of reducing uric acid, including 100 parts of alfalfa, 50 parts of eucommia ulmoides leaves, 50 parts of Sparassis crispa, 15 parts of Ganoderma lucidum, 10.32 parts of cellulase, 3.44 parts of pectinase, 3.44 parts of papain, 0.129 parts of Lactobacillus rhamnosus, and 0.086 parts of Bifidobacterium animalis subsp. lactis; among them, the enzyme activity of cellulase is 60,000 U / g, the enzyme activity of pectinase is 40,000 U / g, the enzyme activity of papain is 100,000 U / g, the viable count of Lactobacillus rhamnosus is 300 billion CFU / g, and the viable count of Bifidobacterium animalis subsp. lactis is 300 billion CFU / g.

[0043] Its preparation method is as follows:

[0044] (1) Crush alfalfa, eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum into powders. Take 100 parts of alfalfa, 50 parts of eucommia ulmoides leaves, 50 parts of Sparassis crispa, and 15 parts of Ganoderma lucidum, and mix them evenly with 2150 parts of water to obtain a mixed solution;

[0045] (2) After adjusting the pH of the mixed solution to 4.5, add 10.32 parts of cellulase, 3.44 parts of pectinase, and 3.44 parts of papain, and carry out enzymatic hydrolysis at a temperature of 50 °C for 75 min to obtain an enzymatic hydrolysate;

[0046] (3) Adjust the pH of the enzymatic hydrolysate to 6.2, raise the temperature of the enzymatic hydrolysate to 118 °C, and carry out enzyme inactivation extraction treatment for 60 min to obtain an enzyme-inactivated extraction solution;

[0047] (4) Wait for the temperature of the enzyme-inactivated extraction solution to drop to about 40 °C, add 0.129 parts of Lactobacillus rhamnosus and 0.086 parts of Bifidobacterium animalis subsp. lactis, and ferment at a temperature of 40 °C for 42 h to obtain a fermentation broth;

[0048] (5) Carry out centrifugation on the fermentation broth, and the supernatant is the fermentation supernatant; concentrate the fermentation supernatant under reduced pressure and dry it in vacuum at 60 °C to obtain a fermentation powder.

[0049] Example 2

[0050] This example provides a fermented product of medicine and food homology with the effect of reducing uric acid, including 80 parts of alfalfa, 65 parts of eucommia ulmoides leaves, 35 parts of Sparassis crispa, 10 parts of Ganoderma lucidum, 8 parts of cellulase, 6 parts of pectinase, 2 parts of papain, 0.11 parts of Lactobacillus rhamnosus, and 0.08 parts of Bifidobacterium animalis subsp. lactis; among them, the enzyme activity of cellulase is 60,000 U / g, the enzyme activity of pectinase is 40,000 U / g, the enzyme activity of papain is 100,000 U / g, the viable count of Lactobacillus rhamnosus is 300 billion CFU / g, and the viable count of Bifidobacterium animalis subsp. lactis is 300 billion CFU / g.

[0051] Its preparation method is as follows:

[0052] (1) Grind alfalfa, eucommia leaves, hydrangea, and ganoderma into powder, and mix 80 parts of alfalfa, 65 parts of eucommia leaves, hydrangea, 35 parts of ganoderma, and 10 parts of ganoderma with 2280 parts of water to obtain a mixed solution;

[0053] (2) After adjusting the pH of the mixed solution to 4, 8 parts of cellulase, 6 parts of pectinase and 2 parts of papain were added, and enzymolysis was performed at 40°C for 90 minutes to obtain an enzymolysis solution;

[0054] (3) adjusting the pH of the enzymatic hydrolysate to 6.5, raising the temperature of the enzymatic hydrolysate to 121° C., and performing enzyme inactivation extraction treatment for 35 minutes to obtain an enzyme inactivation extract;

[0055] (4) When the temperature of the enzyme-killing extract drops to about 42° C., add 0.11 parts of Lactobacillus rhamnosus and 0.08 parts of Bifidobacterium animalis subsp. lactis, and ferment at 42° C. for 18 h to obtain a fermentation liquid;

[0056] (5) The fermentation liquid is centrifuged to obtain the supernatant, which is the fermentation liquid. The fermentation liquid is concentrated under reduced pressure and vacuum dried at 80°C to obtain the fermentation powder.

[0057] Example 3

[0058] The present embodiment provides a medicinal and edible fermented product with uric acid-lowering effect, comprising 110 parts of alfalfa, 35 parts of eucommia leaves, 55 parts of hydrangea, 20 parts of ganoderma, 12 parts of cellulase, 2 parts of pectinase and 3 parts of papain, 0.08 parts of Lactobacillus rhamnosus, and 0.12 parts of Bifidobacterium animalis subsp. lactis; wherein the enzymatic activity of cellulase is 60,000 U / g, the enzymatic activity of pectinase is 40,000 U / g, the enzymatic activity of papain is 100,000 U / g, the viable bacterial count of Lactobacillus rhamnosus is 300 billion CFU / g, and the viable bacterial count of Bifidobacterium animalis subsp. lactis is 300 billion CFU / g.

[0059] Its preparation method is:

[0060] (1) Grinding alfalfa, eucommia leaves, hydrangea, and ganoderma into powder, taking 110 parts of alfalfa, 35 parts of eucommia leaves, 55 parts of hydrangea, 20 parts of ganoderma and 2640 parts of water and mixing them evenly to obtain a mixed solution;

[0061] (2) After adjusting the pH of the mixed solution to 5, 12 parts of cellulase, 2 parts of pectinase and 3 parts of papain were added, and enzymolysis was performed at a temperature of 60°C for 40 minutes to obtain an enzymolysis solution;

[0062] (3) adjusting the pH of the enzymatic hydrolysate to 5.5, raising the temperature of the enzymatic hydrolysate to 110° C., and performing enzyme inactivation extraction treatment for 70 minutes to obtain an enzyme inactivation extract;

[0063] (4)When the temperature of the enzyme-inactivated extraction solution drops to about 35°C, add 0.08 parts of Lactobacillus rhamnosus and 0.12 parts of Bifidobacterium animalis subsp. lactis, and ferment at 35°C for 48 h to obtain a fermentation broth.

[0064] (5)Centrifuge the fermentation broth, and the supernatant is the fermentation clear liquid; concentrate the fermentation clear liquid under reduced pressure and vacuum-dry it at 65°C to obtain a fermentation powder.

[0065] Example 4

[0066] This example provides a medicated and edible ferment with the effect of reducing uric acid, including 120 parts of alfalfa, 40 parts of eucommia leaves, 60 parts of Sparassis crispa, 13 parts of Ganoderma lucidum, 9 parts of cellulase, 5 parts of pectinase, 4 parts of papain, 0.15 parts of Lactobacillus rhamnosus, and 0.07 parts of Bifidobacterium animalis subsp. lactis; among them, the enzyme activity of cellulase is 60,000 U / g, the enzyme activity of pectinase is 40,000 U / g, the enzyme activity of papain is 100,000 U / g, the viable count of Lactobacillus rhamnosus is 300 billion CFU / g, and the viable count of Bifidobacterium animalis subsp. lactis is 300 billion CFU / g.

[0067] Its preparation method is as follows:

[0068] (1)Crush alfalfa, eucommia leaves, Sparassis crispa, and Ganoderma lucidum into powders. Take 120 parts of alfalfa, 40 parts of eucommia leaves, 60 parts of Sparassis crispa, and 13 parts of Ganoderma lucidum and mix them evenly with 2330 parts of water to obtain a mixed solution.

[0069] (2)After adjusting the pH of the mixed solution to 4.5, add 9 parts of cellulase, 5 parts of pectinase, and 4 parts of papain, and carry out enzymatic hydrolysis at 45°C for 60 min to obtain an enzymatic hydrolysate.

[0070] (3)Adjust the pH of the enzymatic hydrolysate to 6.0, raise the temperature of the enzymatic hydrolysate to 115°C, and carry out enzyme-inactivated extraction treatment for 55 min to obtain an enzyme-inactivated extraction solution.

[0071] (4)When the temperature of the enzyme-inactivated extraction solution drops to about 38°C, add 0.15 parts of Lactobacillus rhamnosus and 0.07 parts of Bifidobacterium animalis subsp. lactis, and ferment at 38°C for 36 h to obtain a fermentation broth.

[0072] (5)Centrifuge the fermentation broth, and the supernatant is the fermentation clear liquid; concentrate the fermentation clear liquid under reduced pressure and vacuum-dry it at 70°C to obtain a fermentation powder.

[0073] Example 5

[0074] This example provides a fermented product of medicine and food homology with the effect of reducing uric acid, which includes 90 parts of alfalfa, 60 parts of eucommia ulmoides leaves, 30 parts of Sparassis crispa, 18 parts of Ganoderma lucidum, 11 parts of cellulase, 4 parts of pectinase, 2 parts of papain, 0.1 part of Lactobacillus rhamnosus, and 0.1 part of Bifidobacterium animalis subsp. lactis; among them, the enzyme activity of cellulase is 60,000 U / g, the enzyme activity of pectinase is 40,000 U / g, the enzyme activity of papain is 100,000 U / g, the viable count of Lactobacillus rhamnosus is 300 billion CFU / g, and the viable count of Bifidobacterium animalis subsp. lactis is 300 billion CFU / g.

[0075] Its preparation method is as follows:

[0076] (1) Crush alfalfa, eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum into powders, take 90 parts of alfalfa, 60 parts of eucommia ulmoides leaves, 30 parts of Sparassis crispa, 18 parts of Ganoderma lucidum and mix them evenly with 2178 parts of water to obtain a mixed solution;

[0077] (2) After adjusting the pH of the mixed solution to 4.8, add 11 parts of cellulase, 4 parts of pectinase and 2 parts of papain, and carry out enzymatic hydrolysis at a temperature of 55 °C for 80 min to obtain an enzymatic hydrolysate;

[0078] (3) Adjust the pH of the enzymatic hydrolysate to 5.8, raise the temperature of the enzymatic hydrolysate to 120 °C, and carry out enzyme inactivation extraction treatment for 45 min to obtain an enzyme-inactivated extraction solution;

[0079] (4) Wait for the temperature of the enzyme-inactivated extraction solution to drop to about 37 °C, add 0.1 part of Lactobacillus rhamnosus and 0.1 part of Bifidobacterium animalis subsp. lactis, and ferment at a temperature of 37 °C for 24 h to obtain a fermentation broth;

[0080] (5) Carry out centrifugation treatment on the fermentation broth, and the supernatant is the fermentation supernatant; concentrate the fermentation supernatant under reduced pressure and vacuum dry it at 75 °C to obtain a fermentation powder.

[0081] Example 6

[0082] This example provides a fermented product of medicine and food homology with the effect of reducing uric acid. The difference between this example and Example 1 is that the medicine and food homologous substances are increased by astragalus membranaceus, specifically as follows: 100 parts of alfalfa, 50 parts of eucommia ulmoides leaves, 50 parts of Sparassis crispa, 15 parts of Ganoderma lucidum and 5 parts of astragalus membranaceus, and the rest is the same as in Example 1. And the total weight of astragalus membranaceus, alfalfa, eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum is the same as the total weight of alfalfa, eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum in Example 1.

[0083] Example 7

[0084] This example provides a fermented product of medicated and edible homologous substances with the effect of reducing uric acid. The difference between this example and Example 1 is that the medicated and edible homologous substances are added with Astragalus membranaceus, specifically as follows: 100 parts of alfalfa, 50 parts of Eucommia ulmoides leaves, 50 parts of Sparassis crispa, 15 parts of Ganoderma lucidum, and 8 parts of Astragalus membranaceus, and the rest is the same as in Example 1. And the total weight of Astragalus membranaceus, alfalfa, Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum is the same as the total weight of alfalfa, Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum in Example 1.

[0085] Example 8

[0086] This example provides a fermented product of medicated and edible homologous substances with the effect of reducing uric acid. The difference between this example and Example 1 is that the medicated and edible homologous substances are added with Astragalus membranaceus, specifically as follows: 100 parts of alfalfa, 50 parts of Eucommia ulmoides leaves, 50 parts of Sparassis crispa, 15 parts of Ganoderma lucidum, and 10 parts of Astragalus membranaceus, and the rest is the same as in Example 1. And the total weight of Astragalus membranaceus, alfalfa, Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum is the same as the total weight of alfalfa, Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum in Example 1.

[0087] Comparative Example 1

[0088] This comparative example provides a fermented product of medicated and edible homologous substances. The difference between this comparative example and Example 1 is that it does not include Sparassis crispa, specifically as follows: 100 parts of alfalfa, 50 parts of Eucommia ulmoides leaves, and 15 parts of Ganoderma lucidum, and the rest is the same as in Example 1. And the total weight of alfalfa, Eucommia ulmoides leaves, and Ganoderma lucidum is the same as the total weight of alfalfa, Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum in Example 1.

[0089] Comparative Example 2

[0090] This comparative example provides a fermented product of medicated and edible homologous substances. The difference between this comparative example and Example 1 is that it does not include Eucommia ulmoides leaves, specifically as follows: 100 parts of alfalfa, 50 parts of Sparassis crispa, and 15 parts of Ganoderma lucidum, and the rest is the same as in Example 1. And the total weight of alfalfa, Sparassis crispa, and Ganoderma lucidum is the same as the total weight of alfalfa, Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum in Example 1.

[0091] Comparative Example 3

[0092] This comparative example provides a fermented product of medicated and edible homologous substances. The difference between this comparative example and Example 1 is that it does not include Ganoderma lucidum, specifically as follows: 100 parts of alfalfa, 50 parts of Eucommia ulmoides leaves, and 50 parts of Sparassis crispa, and the rest is the same as in Example 1. And the total weight of alfalfa, Eucommia ulmoides leaves, and Sparassis crispa is the same as the total weight of alfalfa, Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum in Example 1.

[0093] Comparative Example 4

[0094] This comparative example provides a fermented product of medicine and food homology. The difference between this comparative example and Example 1 is that it does not include alfalfa, specifically as follows: 50 parts of Eucommia ulmoides leaves, 50 parts of Sparassis crispa, and 15 parts of Ganoderma lucidum, and the rest is the same as in Example 1. And the total weight of Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum is the same as the total weight of alfalfa, Eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum in Example 1.

[0095] Comparative Example 5

[0096] This comparative example provides a fermented product of medicine and food homology. The difference between this comparative example and Example 1 is that Eucommia ulmoides leaves are replaced by Eucommia ulmoides, specifically as follows: 100 parts of alfalfa, 50 parts of Eucommia ulmoides, 50 parts of Sparassis crispa, and 15 parts of Ganoderma lucidum, and the rest is the same as in Example 1.

[0097] Comparative Example 6

[0098] This comparative example provides a fermented product of medicine and food homology. The difference between this comparative example and Example 1 is that the fermentation agent only includes 0.215 parts of Lactobacillus rhamnosus. The rest is the same as in Example 1.

[0099] Comparative Example 7

[0100] This comparative example provides a fermented product of medicine and food homology. The difference between this comparative example and Example 1 is that the fermentation agent only includes 0.215 parts of Bifidobacterium animalis subsp. lactis. The rest is the same as in Example 1.

[0101] Experimental Example 1 Effect of the fermented product on uric acid in zebrafish

[0102] 1. Experimental animals and breeding environment

[0103] 5 dpf (day past fertilization) wild-type AB strain zebrafish (purchased from Hangzhou Huante Biotechnology Co., Ltd.), raised according to the environmental requirements for the quality control of experimental fish in GB / T 39649-2020, ordinary grade, the breeding system uses reverse osmosis water, the water temperature is maintained at 28 °C, the conductivity is 300 - 500 μS / cm; the pH is 6.8 - 7.5; the dissolved oxygen ≥ 5 mg / L, and the day-night alternation time is 14h / 10h.

[0104] 2. Determination of the maximum tolerable concentration of zebrafish

[0105] Randomly select 5 dpf wild-type AB strain zebrafish and place 30 zebrafish in each well of a 24-well plate. Prepare aqueous solutions of the fermentation powders obtained in Examples 1-8 and Comparative Examples 1-7 with mass concentrations of 125, 250, 500, 1000, 2000, and 4000 μg / mL respectively using fish-raising water. Except for the blank control group, the other groups were administered 2 mL of the aqueous solutions of the fermentation powders of Examples 1-8 and Comparative Examples 1-7 at different concentrations, and the blank control group was given 2 mL of fish-raising water. Each group was set up with 3 parallels. After treatment in an incubator at 28°C for 24 h, observe and record the status and mortality of zebrafish in each group.

[0106] 3. Zebrafish modeling, grouping, and administration

[0107] Randomly select 1530 5 dpf wild-type AB strain zebrafish and randomly divide them into a blank control group, a model control group, and experimental groups (aqueous solutions of the fermentation powders of Examples 1-8 and Comparative Examples 1-7 at the same concentration), with 30 zebrafish in each group. Place them in a 24-well plate, with 3 parallels in each group. Except for the blank control group, the other groups were treated with 250 μmol / L potassium oxonate and 10 μmol / L sodium xanthine to induce the establishment of a hyperuricemia model in 5 dpf zebrafish at 28°C for 24 h.

[0108] After modeling, the experimental groups were given the corresponding aqueous solutions of the fermentation powders at 1000 μg / mL, and the blank control group and the model control group were given an equal amount of fish-raising water, and cultured in an incubator at 28°C for 24 h.

[0109] Potassium oxonate, product number P422648, was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; sodium xanthine, product number X3627, was purchased from Sigma Company, USA.

[0110] 4. Index detection

[0111] 4.1 Uric acid fluorescence value: After culturing the zebrafish in the blank control group, the model control group, and the experimental groups in an incubator at 28°C for 24 h, operate according to the instructions of the uric acid kit, collect data using a multifunctional microplate reader, and analyze the intensity of the uric acid fluorescence signal in zebrafish.

[0112] Amplex TM Red Uric Acid / Urate Oxidase Assay Kit, product number A22181, was purchased from Thermo Fisher Scientific (China) Co., Ltd.

[0113] 4.2 Determine the relative expression levels of OAT1 and HPRT1 genes

[0114] After culturing zebrafish in the blank control group, model control group, and experimental group in an incubator at 28 °C for 24 h, total RNA of zebrafish in each group was extracted using a rapid RNA extraction kit. The concentration of total RNA was measured by absorbance at 260 nm, and the purity of RNA was determined by the ratio of A260 nm / A280 nm.

[0115] For each group, 2 μg of total RNA of zebrafish samples was taken, and cDNA was synthesized using the FastQuant RT kit (primer sequences are shown in Table 1). The expressions of β-actin, HPRT1, and OAT1 genes were detected by q-PCR. β-actin was used as an internal reference for gene expression to calculate the relative RNA expression levels of OAT1 and HPRT1 genes.

[0116] The rapid RNA extraction kit was purchased from Shanghai Yishan Technology Co., Ltd., and the FastKing cDNA First Strand Synthesis Kit was purchased from Beijing Tiangen Biochemical Technology Co., Ltd.

[0117] Table 1 Primer sequence information

[0118]

[0119] 5. Experimental results

[0120] 5.1 Determination results of the maximum tolerable concentration of zebrafish

[0121] The effects of aqueous solutions of baking powder of Examples 1-8 and Comparative Examples 1-7 at different concentrations on the survival of zebrafish are shown in Table 2. It was found that when the concentration of the experimental group was low, it had no effect on the survival of zebrafish. However, as the concentration increased, zebrafish death occurred in each group: zebrafish death occurred at a concentration of 4000 μg / mL in Examples 2 and 3 and Comparative Example 4, while zebrafish death occurred at a concentration of 2000 μg / mL in Examples 1, 4-8 and Comparative Examples 1-3, 5-7. Therefore, aqueous solutions of baking powder corresponding to each group at a concentration of 1000 μg / mL were selected for uric acid-lowering experiment detection on zebrafish.

[0122] Table 2 Mortality rate of zebrafish in the experimental group at different concentrations

[0123]

[0124] 5.2 Evaluation of uric acid-lowering activity

[0125] The effects of aqueous solutions of baking powder in each group on uric acid lowering in zebrafish are shown in Table 3. Compared with the blank control group, the fluorescence signal intensity of the model control group was significantly enhanced, indicating that the high uric acid model of zebrafish was successfully established.

[0126] Compared with the model control group, the fluorescence signal intensity in Examples 1-8 was significantly reduced, indicating that the fermented product obtained by enzymatic hydrolysis and fermentation of the raw materials selected in the present invention has the effect of reducing uric acid.

[0127] Compared with Example 1, the uric acid fluorescence intensity in Comparative Examples 1-5 was significantly enhanced and significant, indicating that the synergistic combination of the raw materials alfalfa, eucommia ulmoides leaves, Sparassis crispa, and Ganoderma lucidum selected in the present invention can achieve the optimal uric acid-lowering effect.

[0128] Compared with Example 1, the uric acid signal intensity in Comparative Examples 6-7 was significantly increased and significant, indicating that the fermentation strains selected in the present invention are the optimal combination.

[0129] Compared with Example 1, there were significant differences in the uric acid fluorescence signal intensity in Examples 6-8, and the fluorescence signal in Examples 6-8 was lower than that in Example 1, indicating that the addition of astragalus enhanced the synergistic effect between the raw materials and better played the role of reducing uric acid.

[0130] Table 3 Evaluation of uric acid-lowering efficacy (n = 3)

[0131]

[0132] Note: Compared with the blank control group, +P < 0.05, ++P < 0.01; compared with the model control group, #P < 0.05, ##P < 0.01; compared with Example 1, △P < 0.05, △△P < 0.01.

[0133] 5.3 Effects on the expression levels of uric acid-lowering related genes

[0134] Hypoxanthine guanine-phosphoribosyltransferases 1 (HPRT1) is a key enzyme in the salvage synthesis pathway of purine metabolism. When this enzyme is lacking, purines are decomposed, leading to an increase in uric acid levels; Organic anion transporter 1 (OAT1) is a member of the SLC22A transporter family and plays an important role in regulating basolateral uric acid excretion.

[0135] The effects of each group on the relative expression levels of OAT1 and HPRT1 genes in zebrafish are shown in Table 4. From the results, it can be seen that: compared with the blank control group, the expression levels of OAT1 and HPRT1 genes in the model control group were significantly decreased, indicating that the model was successfully established; compared with the model control group, the expression levels of OAT1 and HPRT1 genes in Examples 1-8 were significantly increased, indicating that the uric acid-lowering fermented product provided by the present invention can reduce the uric acid concentration in zebrafish by upregulating the expression of OAT1 and HPRT1 genes.

[0136] Compared with Example 1, the expression levels of OAT1 and HPRT1 genes in Comparative Examples 1-7 decreased significantly, indicating that the raw materials and fermentation strains selected in the uric acid-lowering ferment provided by the present invention are an optimal combination. Compared with Example 1, the expression levels of OAT1 and HPRT1 genes in Examples 6-8 increased significantly, indicating that the addition of Astragalus membranaceus improved the expression levels of OAT1 and HPRT1 genes, promoted uric acid excretion, and better exerted the effect of lowering uric acid.

[0137] Table 4 Expression levels of uric acid-lowering related genes (n = 3)

[0138]

[0139] Note: Compared with the blank control group, +P < 0.05, ++P < 0.01; compared with the model control group, #P < 0.05, ##P < 0.01; compared with Example 1, △P < 0.05, △△P < 0.01.

[0140] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than limiting the protection scope of the present invention. Any simple modification or equivalent replacement made by those of ordinary skill in the art to the technical solution of the present invention shall not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A food-medicinal fermented product having the effect of lowering uric acid, characterized in that: The invention comprises the following components in parts by weight: 80-120 parts of clover, 35-65 parts of eucommia leaves, 30-60 parts of hydrangea and 10-20 parts of ganoderma.

2. The food-medicine fermented product with uric acid-lowering effect according to claim 1, characterized in that: The invention comprises the following components in parts by weight: 90-110 parts of clover, 40-60 parts of eucommia leaves, 35-55 parts of hydrangea and 10-15 parts of ganoderma.

3. The food-medicine fermented product with uric acid-lowering effect according to claim 1 or 2, characterized in that: The invention also comprises 0.15-0.27 parts of leavening agent and 12-22 parts of enzyme preparation.

4. The food-medicine fermented product with uric acid-lowering effect according to claim 3, characterized in that: The fermentation agent comprises 0.08-0.15 parts of Lactobacillus rhamnosus and 0.07-0.12 parts of Bifidobacterium animalis subsp. lactis; The enzyme preparation comprises 8-12 parts of cellulase, 2-6 parts of pectinase and 2-4 parts of papain.

5. The food-medicine fermented product with uric acid-lowering effect according to any one of claims 1 to 4, characterized in that: Also includes 5-10 portions of Astragalus.

6. The method for preparing the edible-medicinal fermented product with uric acid lowering effect according to any one of claims 1 to 5, characterized in that: The steps include: (1) mixing the raw materials with water to obtain a mixed solution; (2) adjusting the pH of the mixed solution to 4-5, adding an enzyme preparation, and performing enzymatic hydrolysis to obtain an enzymatic hydrolyzate; (3) adjusting the pH of the enzymatic hydrolyzate to 5.5-6.5, raising the temperature, and performing enzyme inactivation extraction treatment to obtain an enzyme inactivation extract; (4) adding a fermentation agent to the enzyme-killed extract to carry out fermentation to obtain a fermentation liquid; (5) The fermentation liquid is centrifuged, and the supernatant is the fermentation liquid.

7. The method for preparing the food-medicine fermented product according to claim 6, characterized in that: In the step (2), the enzymolysis temperature is 40-60° C. and the enzymolysis time is 40-90 min.

8. The method for preparing the food-medicine fermented product according to claim 6, characterized in that: The enzyme inactivation extraction temperature in step (3) is 110-121° C., and the enzyme inactivation extraction time is 35-70 min.

9. The method for preparing the food-medicine fermented product according to claim 6, characterized in that: The fermentation temperature of step (4) is 35-42°C and the fermentation time is 18-48h.

10. Use of the food-medicine fermented product according to any one of claims 1 to 5 or the food-medicine fermented product obtained by the preparation method according to any one of claims 6 to 9 in preparing products for treating hyperuricemia and / or gout.

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