Composite peptide dietary supplement for improving qi depression constitution and preparation method, preparation and application thereof

Abstract

The invention provides a composite peptide dietary supplement for improving qi depression constitution as well as a preparation method, a preparation and application of the composite peptide dietary supplement, and belongs to the technical field of food biology. On the first aspect, the invention provides the composite peptide dietary supplement, and the composite peptide dietary supplement comprises the following raw materials: fingered citron, fructus amomi, citron, poria cocos, malt, cordyceps militaris, Chinese dates, liquorice, ginseng, sorbitol, arabinose, tea leaf theanine and mint; on the second aspect, the invention provides a preparation method of the composite peptide dietary supplement; and thirdly, the invention provides a preparation of the composite peptide dietary supplement. In the fourth aspect, the invention also provides application of the composite peptide dietary supplement and the preparation thereof in preparation of food or medicines suitable for people with qi depression constitution.

Description

Technical Field

[0001] The present invention relates to the field of food biotechnology, and in particular to a composite peptide dietary supplement for improving qi stagnation constitution, and a preparation method, preparation and application thereof. Background Art

[0002] With the progress of human society, world medicine is changing from disease medicine to "human" medicine. From health to sub-health and then to disease, the influence of physical factors cannot be ignored. Various physical biases are the inherent basis for the occurrence of diseases. At the same time, it is precisely because of the differences in physical constitution that the occurrence and outcome of body diseases are also different. Human physique is both stable and variable. Through intervention, the imbalanced state of human physique can be improved and adjusted, thereby restoring health. Diet for body identification and improvement of biased physique can not only adjust the sub-health state and block its transformation to the disease state, but also achieve the health care effect of active prevention.

[0003] Constitution in traditional Chinese medicine refers to the comprehensive and relatively stable characteristics of an individual in terms of morphological structure, physiological function and psychological state formed during the life process of birth, growth, maturity and aging, which are manifested in individual differences in structure, function, metabolism and response to external stimuli, susceptibility to certain causes and diseases, and certain tendencies in the transmission and progression of diseases. Epidemiological surveys have found that Qi stagnation constitution is a common biased constitution among the nine types of constitution.

[0004] Qi Depression Constitution is a physical state formed by long-term emotional depression and stagnation of Qi, with introverted and unstable personality, melancholy and fragility, sensitivity and suspicion as the main manifestations. People in this physical state are mostly middle-aged and young people, and more common in women. They are often lonely and introverted, easily sentimental, and narrow-minded. The main disease of people with Qi Depression Constitution is the liver, and also affects the heart, stomach, large intestine, and small intestine. It is easy to hurt emotions and diet, and it is easy to produce Qi stagnation, such as depression, insomnia, plum pit qi, panic, etc. Modern research shows that this kind of constitution is prone to tumors. Conditioning treatment should regulate emotions and dredge Qi. The term Qi depression comes from "Suwen·Liuyuan Zhengji Dalun", which is mostly caused by emotional discomfort and Qi stagnation. "Dan Xi's Heart Method·Six Depressions": "Those with qi stagnation have chest and flank pain, and a deep and sluggish pulse." "Zhengzhi Huibu·Depression and Symptoms": "Qi stagnation causes chest fullness and flank pain, belching and abdominal distension." People with qi stagnation constitution are often unhappy and low in mood, easily nervous and anxious, sentimental, emotionally fragile, easily afraid or frightened, have pain in the ribs or breasts, sigh for no reason, and have a depressed look.

[0005] Causes of Qi stagnation and common diseases: 1. Congenital inheritance. 2. Staying up late frequently. 3. Liver is the main cause of the disease, and the heart, stomach, large intestine and small intestine are also affected. Emotions and diet are easily hurt, which can easily lead to poor Qi. Long-term excessive stress and excessive thinking. 4. Sudden mental stimulation, such as the death of a loved one, sudden fright, etc. 5. Common diseases include peptic ulcer, neurosis, insomnia, bipolar disorder, menopausal syndrome, depression, insomnia, plum pit qi, panic, etc.

[0006] The constitution theory believes that "body and disease are related". Constitution is not only an internal factor in the occurrence of disease, but also affects the development and outcome of the disease.

[0007] At present, no candy health food for people with qi stagnation constitution using bergamot as raw materials has been found in the market. In view of this, the present invention formulates a formula according to the standard of "Classification and Determination of Chinese Medical Constitution" officially released by the Chinese Association of Traditional Chinese Medicine on April 9, 2009, selects raw materials that are both medicinal and edible, and adopts scientific technology to make concentrated small molecule peptide tablet candy. The candy has the effects of soothing the liver and regulating qi, promoting qi and strengthening the spleen, and can achieve the effect of regulating the sub-health state of people with qi stagnation constitution.

[0008] Therefore, the present invention provides a dietary supplement of bergamot, amomum villosum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, sorbitol, arabinose, tea theanine, and mint composite peptide, and a preparation method, preparation, and application thereof. Summary of the invention

[0009] The object of the present invention is to provide a composite peptide dietary supplement for improving qi stagnation constitution and a preparation method, preparation and application thereof.

[0010] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0011] In a first aspect, the present invention provides a composite peptide dietary supplement for improving qi stagnation constitution, wherein the raw materials of the composite peptide dietary supplement include bergamot, amomum villosum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, sorbitol, arabinose, and tea theanine.

[0012] Furthermore, the raw material of the composite peptide dietary supplement also includes mint.

[0013] Furthermore, the raw materials of the composite peptide dietary supplement are, by weight, 2-10 parts of bergamot, 2-10 parts of amomum, 2-10 parts of citron, 2-8 parts of poria, 2-8 parts of malt, 2-8 parts of cordyceps militaris, 2-8 parts of jujube, 2-8 parts of licorice, 2-8 parts of ginseng, 2-4 parts of sorbitol, 2-4 parts of arabinose, 1-3 parts of tea theanine, and 2-8 parts of mint.

[0014] Furthermore, the raw materials of the composite peptide dietary supplement are, by weight, 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint; or 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 4 parts of jujube, 4 parts of licorice, 4 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 4 parts of mint.

[0015] In a second aspect, the present invention provides a method for preparing a composite peptide dietary supplement for improving qi stagnation constitution, comprising:

[0016] The method comprises the following steps: the first step is to prepare the original pulp; the second step is to prepare the fermentation agent; the third step is to prepare the diluted mixed pulp; the fourth step is to prepare the fermentation liquid; the fifth step is the yeast autolysis and ultrasonic wall breaking; the sixth step is the ultrafiltration; the seventh step is the purification; and the eighth step is the drying.

[0017] Furthermore, the specific process for preparing the stock pulp in the first step is to take bergamot, amomum villosum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint, wash them separately, soak them until there is no dry core, drain the surface water, and crush them into stock pulp for standby use.

[0018] Furthermore, the specific process of preparing the yeast agent in the second step is to select high-temperature resistant high-activity yeast and Aspergillus oryzae for mixed culture to form a binary yeast solution A, and to mix and culture three kinds of bacteria, Lactobacillus bulgaricus, Streptococcus thermophilus, and Lactobacillus acidophilus, to form a yeast solution B for standby use.

[0019] Furthermore, the specific process for preparing the diluted mixed slurry in the third step is to use the weight of the original slurry obtained in the first step as the standard, add sorbitol, arabinose, tea theanine, an appropriate amount of pure water, and an appropriate amount of brown sugar to the original slurry in the fermentation tank, keep boiling for a period of time, stop heating, cool down, and obtain a diluted mixed slurry for standby use.

[0020] Furthermore, the specific process for preparing the fermentation liquid in the fourth step is to inoculate the binary yeast liquid A and yeast liquid B in the second step into the diluted mixed slurry obtained in the third step at a ratio of 0.02 to 0.06 times by weight.

[0021] Furthermore, the specific process of the fifth step of yeast autolysis and ultrasonic cell wall breaking is to control appropriate yeast autolysis conditions after terminating the fermentation, autolyze the yeast in the fermentation broth and use ultrasonic cell wall breaking extraction technology to obtain rich yeast small molecule peptides and amino acids.

[0022] Furthermore, the specific process of the sixth step ultrafiltration is to take the supernatant and use an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons for membrane separation to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0023] Furthermore, the specific process of the seventh step of purification is purification using 732 type cation exchange resin.

[0024] Furthermore, the specific process of the eighth step of drying is to freeze-dry the purified product to obtain the active polypeptide or spray-dry it.

[0025] Furthermore, in the second step, high-activity yeast and Aspergillus oryzae are inoculated into a culture medium at a temperature of 30 to 38° C., cultured at 160 rpm for 12 to 48 hours to prepare a binary yeast solution A; Lactobacillus bulgaricus, Streptococcus thermophilus and Lactobacillus acidophilus are inoculated into a culture medium at a temperature of 30 to 42° C., cultured for 5 to 20 hours to prepare a yeast solution B.

[0026] Furthermore, in the third step, based on the weight of the raw pulp obtained in the first step, sorbitol, arabinose, tea theanine, 2 to 5 times of pure water, and 0.1 to 0.2 times of brown sugar are added to the raw pulp in the fermentation tank, and the mixture is boiled for 5 minutes, and then the heating is stopped and the mixture is cooled to obtain a diluted mixed pulp for standby use.

[0027] Furthermore, in the fourth step, the fermentation time is 1 to 2 days.

[0028] Furthermore, in the second step, the culture medium is 1% yeast extract powder, 75-79% water, 3-5% brown sugar and 15-20% potato.

[0029] In a third aspect, the present invention provides a preparation of the composite peptide dietary supplement, which can be a tablet, an granule or an oral solution.

[0030] In a fourth aspect, the present invention also provides a use of the composite peptide dietary supplement and the preparation thereof in preparing food or medicine suitable for people with qi stagnation constitution.

[0031] Compared with the prior art, the beneficial effects of the present invention include:

[0032] Compared with the prior art, the composite peptide dietary supplement provided by the present invention has a more scientific and reasonable formula, a high-efficiency and low-cost preparation method, high active peptide activity and high yield, high preparation bioavailability, and a significant effect on improving qi stagnation constitution, with significant improvements in many aspects. DETAILED DESCRIPTION

[0033] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. If specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0034] The features and properties of the present invention are further described in detail below in conjunction with the embodiments:

[0035] Example 1

[0036] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of tuckahoe, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, and 2 parts of tea theanine. Wash bergamot, amomum, citron, tuckahoe, malt, cordyceps militaris, jujube, licorice, and ginseng separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0037] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato. Select activated high temperature resistant high activity yeast and Aspergillus oryzae in the inoculation ratio of 1:1, the inoculation amount is 2.5%, 160 rpm, 34℃ mixed culture for 30h, to make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus in the inoculation ratio of 1:1:1, the inoculation amount is 2%, 36℃ mixed culture for 12h, to make fermentation liquid B, and set aside.

[0038] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0039] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0040] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0041] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0042] Step 7: Purification: Purification using 732 type cation exchange resin.

[0043] Step 8: Drying: The purified product is spray dried.

[0044] Example 2

[0045] The first step is to prepare the stock solution: take 10 parts of bergamot, 2 parts of amomum, 10 parts of citron, 2 parts of poria, 8 parts of malt, 2 parts of cordyceps militaris, 8 parts of jujube, 2 parts of licorice, 8 parts of ginseng, 2 parts of sorbitol, 4 parts of arabinose, 1 part of tea theanine, and 2 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0046] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 79% water, 3% brown sugar and 17% potato. Select activated high temperature resistant high activity yeast and Aspergillus oryzae in the inoculation ratio of 1:1, the inoculation amount is 2%, 200 rpm, 30℃ mixed culture for 48h, and make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus in the inoculation ratio of 1:1:1, the inoculation amount is 1%, and 42℃ mixed culture for 5h to make fermentation liquid B for standby use.

[0047] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 5 times pure water, and 0.1 times brown sugar to the original slurry in the fermentation tank, boil it for 7 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0048] The fourth step is to prepare the fermentation liquid: inoculate the binary yeast liquid A and yeast liquid B in the second step into the diluted mixed slurry obtained in the third step at a ratio of 0.03 times by weight, respectively, at a fermentation temperature of 36° C. and a fermentation time of 24 hours.

[0049] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 57°C for 12 hours, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1.5 minutes, rest for 2 minutes, and breaking for 10 times.

[0050] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0051] Step 7: Purification: Purification using 732 type cation exchange resin.

[0052] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0053] Example 3

[0054] The first step is to prepare the stock solution: 2 parts of bergamot, 10 parts of amomum, 2 parts of citron, 8 parts of poria, 2 parts of malt, 8 parts of cordyceps militaris, 2 parts of jujube, 8 parts of licorice, 2 parts of ginseng, 4 parts of sorbitol, 2 parts of arabinose, 3 parts of tea theanine, and 8 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, and crush into stock solution for standby use.

[0055] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 75% water, 5% brown sugar and 19% potato. Select activated high temperature resistant high activity yeast and Aspergillus oryzae in the inoculation ratio of 1:1, the inoculation amount is 3%, 120 rpm, 38℃ mixed culture for 12h, and make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus in the inoculation ratio of 1:1:1, the inoculation amount is 3%, and 30℃ mixed culture for 20h to make fermentation liquid B for standby use.

[0056] The third step is to prepare the diluted mixed slurry: using the weight of the original slurry obtained in the first step as the standard, add sorbitol, arabinose, tea theanine, 2 times of pure water, and 0.2 times of brown sugar to the original slurry in the fermentation tank, boil it for 3 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0057] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.05 times by weight, respectively. The fermentation temperature is 30° C. and the fermentation time is 48 hours.

[0058] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 53°C and 36h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 0.5min, rest for 0.5min, and breaking for 30 times.

[0059] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0060] Step 7: Purification: Purification using 732 type cation exchange resin.

[0061] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0062] Example 4

[0063] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 4 parts of jujube, 4 parts of licorice, 4 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 4 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, and crush into stock solution for standby use.

[0064] The second step is to prepare the fermentation agent: 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato in the culture medium. Select activated high-temperature resistant and highly active yeast and Aspergillus oryzae in a ratio of 1:1, with an inoculation amount of 2.5% respectively, 160 rpm, 34°C mixed culture for 30 hours to make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, and Lactobacillus acidophilus in a ratio of 1:1:1, with an inoculation amount of 2% respectively, 36°C mixed culture for 12 hours to make fermentation liquid B for standby use.

[0065] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0066] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0067] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0068] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0069] Step 7: Purification: Purification using 732 type cation exchange resin.

[0070] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0071] Example 5

[0072] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0073] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato. Select activated high temperature resistant high activity yeast and Aspergillus oryzae in the inoculation ratio of 1:1, the inoculation amount is 2.5%, 160 rpm, 34℃ mixed culture for 30h, to make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus in the inoculation ratio of 1:1:1, the inoculation amount is 2%, 36℃ mixed culture for 12h, to make fermentation liquid B, and set aside.

[0074] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0075] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0076] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0077] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0078] Step 7: Purification: Purification using 732 type cation exchange resin.

[0079] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0080] Comparative Example 1

[0081] The first step is to prepare ultrafine powder: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Crush them into powder with a particle size of about 100 μm and set aside.

[0082] The second step is water extraction method: add 10 times the amount of distilled water, soak for 4 hours, heat to boil, decoct for 2 hours, filter, add 5 times the amount of distilled water to the residue, heat to boil, decoct for 1 hour, filter; mix the two filtrates.

[0083] The third step is to obtain the product: after the filtrate is appropriately concentrated, freeze-drying is performed to obtain the product.

[0084] Comparative Example 2

[0085] The first step is to prepare ultrafine powder: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Crush them into powder with a particle size of about 100 μm and set aside.

[0086] The second step is ethanol extraction method: add 10 times the amount of 95% ethanol, soak for 4 hours, heat under reflux for extraction for 2 hours and filter, add 5 times the amount of 95% ethanol to the residue, heat under reflux for extraction for 2 hours and filter; mix the two filtrates.

[0087] The third step is to obtain the product: after the filtrate is appropriately concentrated, freeze-drying is performed to obtain the product.

[0088] Comparative Example 3

[0089] The first step is to prepare ultrafine powder: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Crush them into powder with a particle size of about 100 μm and set aside.

[0090] The second step is to extract with ethanol first and then with water: add 10 times the amount of 95% ethanol, soak for 4 hours, heat and reflux to extract for 2 hours, filter, add 5 times the amount of 95% ethanol to the residue, heat and reflux to extract for 2 hours, filter; mix the two filtrates. Add 10 times the amount of distilled water to the residue, soak for 4 hours, heat and boil for 2 hours, filter, add 5 times the amount of distilled water to the residue, heat and boil for 1 hour, filter; mix the two filtrates with the alcohol extraction filtrate.

[0091] The third step is to obtain the product: after the filtrate is appropriately concentrated, freeze-drying is performed to obtain the product.

[0092] Comparative Example 4

[0093] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0094] The second step is to prepare the fermentation agent: the culture medium is 40.5g / L glucose, 36.8g / L peptone, 17.3g / L corn steep liquor, 14g / L potassium nitrate, 1.5g / L yeast nutrient salt, 0.6g / L potassium dihydrogen phosphate, and 0.8g / L magnesium sulfate. Select activated high-temperature resistant and highly active yeast and Aspergillus oryzae in an inoculation ratio of 1:1, with an inoculation amount of 2.5% respectively, 160 rpm, 34°C mixed culture for 30h, to prepare binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, and Lactobacillus acidophilus in an inoculation ratio of 1:1:1, with an inoculation amount of 2% respectively, 36°C mixed culture for 12h, to prepare fermentation liquid B for standby use.

[0095] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0096] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0097] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0098] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0099] Step 7: Purification: Purification using 732 type cation exchange resin.

[0100] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0101] Comparative Example 5

[0102] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0103] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato. Select activated Kluyveromyces marxii and Aspergillus oryzae in a ratio of 1:1, the inoculation amount is 2.5%, 160 rpm, 34℃ mixed culture for 30h, to make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus in a ratio of 1:1:1, the inoculation amount is 2%, 36℃ mixed culture for 12h, to make fermentation liquid B, and set aside.

[0104] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0105] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0106] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0107] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0108] Step 7: Purification: Purification using 732 type cation exchange resin.

[0109] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0110] Comparative Example 6

[0111] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0112] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato. Select activated high temperature resistant high activity yeast and Aspergillus oryzae in the inoculation ratio of 1:1, the inoculation amount is 2.5% respectively, 160 rpm, 34℃ mixed culture for 30h, to make binary fermentation liquid A; select activated Bacillus subtilis, thermophilic Streptococcus, Lactobacillus casei inoculation ratio of 1:1:1, the inoculation amount is 2% respectively, 36℃ mixed culture for 12h, to make fermentation liquid B, and set aside.

[0113] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0114] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0115] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0116] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0117] Step 7: Purification: Purification using 732 type cation exchange resin.

[0118] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0119] Comparative Example 7

[0120] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0121] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato. Select activated high temperature resistant high activity yeast and Aspergillus oryzae in the inoculation ratio of 1:1, the inoculation amount is 2.5%, 160 rpm, 34℃ mixed culture for 30h, to make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus in the inoculation ratio of 1:1:1, the inoculation amount is 2%, 36℃ mixed culture for 12h, to make fermentation liquid B, and set aside.

[0122] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of glucose to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0123] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0124] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0125] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0126] Step 7: Purification: Purification using 732 type cation exchange resin.

[0127] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0128] Comparative Example 8

[0129] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0130] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato. Select activated high temperature resistant high activity yeast and Aspergillus oryzae in the inoculation ratio of 1:1, the inoculation amount is 2.5%, 160 rpm, 34℃ mixed culture for 30h, to make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus in the inoculation ratio of 1:1:1, the inoculation amount is 2%, 36℃ mixed culture for 12h, to make fermentation liquid B, and set aside.

[0131] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0132] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0133] The fifth step is ultrasonic wall breaking: ultrasonic wall breaking technology is used, ultrasonic breaking for 1 minute, rest for 1 minute, and breaking 20 times.

[0134] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0135] Step 7: Purification: Purification using 732 type cation exchange resin.

[0136] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0137] Comparative Example 9

[0138] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0139] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato. Select activated high temperature resistant high activity yeast and Aspergillus oryzae in the inoculation ratio of 1:1, the inoculation amount is 2.5%, 160 rpm, 34℃ mixed culture for 30h, to make binary fermentation liquid A; select activated Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus in the inoculation ratio of 1:1:1, the inoculation amount is 2%, 36℃ mixed culture for 12h, to make fermentation liquid B, and set aside.

[0140] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0141] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0142] Step 5: yeast autolysis: After the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, and the yeast autolysis conditions are controlled at 55°C and 24h.

[0143] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0144] Step 7: Purification: Purification using 732 type cation exchange resin.

[0145] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0146] Comparative Example 10

[0147] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0148] The second step is ultrafiltration: taking the weight of the stock solution obtained in the first step as the standard, add sorbitol, arabinose, tea theanine, and 3 times the volume of water to the stock solution and stir it thoroughly, then coarsely filter the supernatant and use an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons for membrane separation to remove large molecular impurities and obtain small molecular peptides below 10,000 Daltons and other components.

[0149] The third step is purification: purification using 732 type cation exchange resin.

[0150] The fourth step is drying: freeze-drying the purified product to obtain active polypeptides.

[0151] Comparative Example 11

[0152] The first step is to prepare the stock solution: take 8 parts of bergamot, 8 parts of amomum, 8 parts of citron, 6 parts of poria, 6 parts of malt, 6 parts of cordyceps militaris, 6 parts of jujube, 6 parts of licorice, 6 parts of ginseng, 3 parts of sorbitol, 3 parts of arabinose, 2 parts of tea theanine, and 6 parts of mint. Wash bergamot, amomum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint separately, soak until there is no dry core, drain the surface water, crush into stock solution, and set aside.

[0153] The second step is to prepare the yeast agent: the culture medium is 2% yeast extract powder, 50% water, 9% brown sugar and 39% potato, and activated Aspergillus oryzae is selected, the inoculation amount is 2%, 31°C, 160 rpm, and aeration culture is carried out for 24 hours to prepare yeast solution A for standby use; the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato, and activated Lactobacillus acidophilus is selected, the inoculation amount is 2%, and closed culture is carried out at 35°C for 48 hours to prepare binary yeast solution B for standby use.

[0154] The third step is to prepare the diluted mixed slurry: based on the weight of the original slurry obtained in the first step, add sorbitol, arabinose, tea theanine, 3.5 times of pure water, and 0.15 times of brown sugar to the original slurry in the fermentation tank, boil it for 5 minutes, stop heating, cool it down, and obtain the diluted mixed slurry for standby use.

[0155] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0156] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0157] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0158] The seventh step is purification: purification using anion exchange resin TEHUAMP-600.

[0159] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0160] Comparative Example 12

[0161] The first step is to prepare the stock solution: take 8 parts of ginseng, 6 parts of wolfberry, 6 parts of hawthorn, 6 parts of chicken gizzard lining, 6 parts of orange peel, 8 parts of peach kernel, and 6 parts of chicory. Wash them separately, soak them until there is no dry core, drain the surface water, crush them into stock solution, and set aside.

[0162] The second step is to prepare the fermentation agent: the culture medium is 2% yeast extract powder, 2% glucose and 5% soybean meal, and the balance is water. The activated Paecilomyces punctatus is inoculated at an inoculation rate of 2%, and aerobically cultured at 160 rpm and 25°C for 30 hours to prepare a binary fermentation liquid A; the activated Bifidobacterium lactis is inoculated at an inoculation rate of 2%, and sealed cultured at 36°C for 12 hours to prepare a fermentation liquid B for standby use.

[0163] The third step is to prepare the diluted mixed slurry: according to the weight of the original slurry obtained in the first step, add 3.5 times of pure water and 0.15 times of brown sugar into the fermentation tank, boil for 5 minutes, stop heating, cool down, and obtain the diluted mixed slurry for standby use.

[0164] Step 4: prepare the fermentation liquid: inoculate the fermentation liquid A and B in the second step into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, and ferment at a closed temperature of 33° C. for 36 hours.

[0165] The fifth step is ultrasonic cell wall breaking: after the fermentation is terminated, ultrasonic cell wall breaking technology is used, ultrasonic breaking for 1 minute, rest for 1 minute, and breaking 20 times.

[0166] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 6000 Daltons to remove macromolecular impurities and obtain components such as polypeptides below 6000 Daltons.

[0167] Step 7: Purification: Purification using 724 cation exchange resin.

[0168] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0169] Comparative Example 13

[0170] The first step is to prepare the stock solution: take 8 parts of ginseng, 6 parts of lotus seeds, 6 parts of Amomum villosum, 8 parts of Chinese yam, 6 parts of Poria cocos, 8 parts of white hyacinth bean, 6 parts of orange peel, 6 parts of licorice, and 6 parts of jujube. Wash them separately, soak them until there is no dry core, drain the surface water, crush them into stock solution, and set aside.

[0171] The second step is to prepare the fermentation agent: the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% soybean meal superfine powder. Select activated Kluyveromyces marxii, inoculated at 2.5%, 160 rpm, 28℃ sealed fermentation for 8 hours, and then aerate and culture for 30 hours to make fermentation liquid A; select activated Lactobacillus bulgaricus, Bifidobacterium longum, and Lactobacillus acidophilus inoculated at a ratio of 1:1:1, inoculated at 2% respectively, and mixed cultured at 36℃ for 12 hours to make fermentation liquid B for standby use.

[0172] The third step is to prepare the diluted mixed slurry: according to the weight of the original slurry obtained in the first step, add 3.5 times of pure water and 0.15 times of brown sugar into the fermentation tank, boil for 5 minutes, stop heating, cool down, and obtain the diluted mixed slurry for standby use.

[0173] The fourth step is to prepare the fermentation liquid: the yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight, respectively. The fermentation temperature is 33° C. and the time is 36 hours.

[0174] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0175] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0176] Step 7: Purification: Purification using Tehua IRA204 anion exchange resin.

[0177] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0178] Comparative Example 14

[0179] The first step is to prepare the stock solution: take 12 parts of sea buckthorn, 8 parts of patchouli, 8 parts of perilla, 6 parts of oats, 6 parts of black plum, 6 parts of houttuynia, 6 parts of mint, 6 parts of angelica, 6 parts of Euryale, 2 parts of red dates, 2 parts of lactitol, 2 parts of isomaltooligosaccharide, and 6 parts of licorice. Wash sea buckthorn, patchouli, perilla, oats, black plum, houttuynia, mint, angelica, Euryale, red dates, and licorice separately, soak until there is no dry core, drain the surface water, and crush into stock solution for standby use.

[0180] The second step is to prepare the yeast agent: the culture medium is 2% yeast extract powder, 50% water, 9% brown sugar and 39% potato, and activated high-temperature resistant and highly active yeast is selected, the inoculation amount is 2%, 31°C, 160 rpm, and aeration culture is carried out for 24 hours to prepare yeast solution A for standby use; the culture medium is 1% yeast extract powder, 77% water, 4% brown sugar and 18% potato, and activated Lactobacillus acidophilus is selected, the inoculation amount is 2%, and closed culture is carried out at 35°C for 48 hours to prepare binary yeast solution B for standby use.

[0181] The third step is to prepare the diluted mixed slurry: according to the weight of the original slurry obtained in the first step, add lactitol, isomaltooligosaccharide, 3.5 times purified water, and 0.15 times brown sugar into the fermentation tank, boil for 5 minutes, stop heating, cool down, and obtain the diluted mixed slurry for standby use.

[0182] The fourth step is to prepare the fermentation liquid: the binary yeast liquid A and the yeast liquid B in the second step are inoculated into the diluted mixed slurry obtained in the third step at a ratio of 0.04 times by weight respectively, the fermentation temperature is 33° C., and the fermentation time is 36 hours.

[0183] The fifth step is yeast autolysis and ultrasonic cell wall breaking: after the fermentation is terminated, the yeast in the fermentation liquid is autolyzed, the yeast autolysis conditions are controlled at 55°C, 24h, and then the ultrasonic cell wall breaking process is adopted, ultrasonic breaking for 1min, rest for 1min, and breaking 20 times.

[0184] The sixth step is ultrafiltration: the supernatant is taken and separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecular peptides and other components below 10,000 Daltons.

[0185] The seventh step is purification: purification using anion exchange resin TEHUAMP-600.

[0186] Step 8: Drying: The purified product is freeze-dried to obtain the active polypeptide.

[0187] Experimental Example 1

[0188] The results are shown in Table 1:

[0189]

[0190]

[0191] As can be seen from Table 1, the polypeptide yield and polypeptide purity of the corresponding products obtained by using bergamot, amomum villosum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, sorbitol, arabinose, tea theanine and mint in the embodiments of the present invention are significantly improved compared with the control groups, and the histamine content is low.

[0192] Experimental Example 2

[0193] 1. Methods

[0194] Rats were randomly divided into a normal group, a model group, each group of the embodiment and each group of the comparative example, with 10 rats in each group. The daily gavage sample dosage of each group of the embodiment and the comparative example was calculated according to the standard animal equivalent dose conversion coefficient method (based on 50g of raw medicinal materials / kg body weight / day for rats). Rats in the normal group and the model group were gavaged with starch of equal weight, twice a day, for 28 consecutive days. On the 29th day, except for the normal group, each group was subcutaneously injected with 0.1% epinephrine, 0.1ml / rat, and rats in each group were killed 24 hours later. The length and dry weight of the in vitro thrombus of each group were measured.

[0195] 2. Results

[0196] Statistical results of in vitro thrombus length and dry weight determination in each group See Table 2.

[0197] Table 2

[0198]

[0199]

[0200] The links not described in detail in the present invention are all common knowledge that can be selected by ordinary technicians in the field. Although the present invention has been described in detail above with general descriptions and specific implementation schemes, it is obvious to ordinary technicians in the field that some modifications or improvements can be made on the basis of the present invention. Therefore, these modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. A composite peptide dietary supplement for improving qi stagnation constitution, characterized in that: The composite peptide dietary supplement comprises the following raw materials: bergamot, amomum villosum, citron, tuckahoe, malt, cordyceps militaris, jujube, licorice, ginseng, sorbitol, arabinose and tea theanine.

2. The composite peptide dietary supplement according to claim 1, characterized in that: The composite peptide dietary supplement raw material also includes mint.

3. The composite peptide dietary supplement according to claim 2, characterized in that: In terms of weight, the bergamot is 2-10 parts, the amomum is 2-10 parts, the citron is 2-10 parts, the poria is 2-8 parts, the malt is 2-8 parts, the cordyceps militaris is 2-8 parts, the jujube is 2-8 parts, the licorice is 2-8 parts, the ginseng is 2-8 parts, the sorbitol is 2-4 parts, the arabinose is 2-4 parts, the tea theanine is 1-3 parts, and the mint is 2-8 parts.

4. The composite peptide dietary supplement according to claim 2, characterized in that: In terms of weight, the bergamot is 8 parts, the Amomum villosum is 8 parts, the citron is 8 parts, the Poria cocos is 6 parts, the malt is 6 parts, the Cordyceps militaris is 6 parts, the jujube is 6 parts, the licorice is 6 parts, the ginseng is 6 parts, the sorbitol is 3 parts, the arabinose is 3 parts, the tea theanine is 2 parts, and the mint is 6 parts; or the bergamot is 8 parts, the Amomum villosum is 8 parts, the citron is 8 parts, the Poria cocos is 6 parts, the malt is 6 parts, the Cordyceps militaris is 6 parts, the jujube is 4 parts, the licorice is 4 parts, the ginseng is 4 parts, the sorbitol is 3 parts, the arabinose is 3 parts, the tea theanine is 2 parts, and the mint is 4 parts.

5. The method for preparing the composite peptide dietary supplement according to any one of claims 1 to 4, characterized in that: The following steps are involved: The first step is to prepare the original slurry; the second step is to prepare the yeast agent; the third step is to prepare the diluted mixed slurry; the fourth step is to prepare the fermentation liquid; the fifth step is to autolyze the yeast and break the wall with ultrasonic wave; the sixth step is to ultrafilter; the seventh step is to purify; and the eighth step is to dry.

6. The method for preparing the composite peptide dietary supplement according to claim 5, characterized in that: The specific process of preparing the stock solution in the first step is to take bergamot, amomum villosum, citron, poria, malt, cordyceps militaris, jujube, licorice, ginseng, and mint, wash them separately, soak them until there is no dry core, drain the surface water, crush them into stock solution, and set aside; The specific process of preparing the fermentation agent in the second step is to select high-temperature resistant high-activity yeast and Aspergillus oryzae to be mixed and cultured to form a binary fermentation liquid A, and to mix and culture three kinds of bacteria, Lactobacillus bulgaricus, Streptococcus thermophilus, and Lactobacillus acidophilus to form a fermentation liquid B for standby use; The specific process of preparing the diluted mixed slurry in the third step is to add sorbitol, arabinose, tea theanine, an appropriate amount of purified water, and an appropriate amount of brown sugar to the raw slurry in the fermenter based on the weight of the raw slurry obtained in the first step, boil it for a period of time, stop heating, cool it down, and obtain the diluted mixed slurry for standby use; The specific process of preparing the fermentation liquid in the fourth step is to inoculate the binary yeast liquid A and yeast liquid B in the second step into the diluted mixed slurry obtained in the third step at a ratio of 0.02 to 0.06 times by weight; The specific process of the fifth step of yeast autolysis and ultrasonic cell wall breaking is to control appropriate yeast autolysis conditions after terminating the fermentation, autolyze the yeast in the fermentation broth and use ultrasonic cell wall breaking extraction process to obtain rich yeast small molecule peptides and amino acids; The specific process of the sixth step of ultrafiltration is to take the supernatant and use an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons for membrane separation to remove macromolecular impurities and obtain small molecular peptides and other components below 10,000 Daltons; The specific process of the seventh step of purification is purification using 732 type cation exchange resin; The specific process of the eighth step of drying is to freeze-dry the purified product to obtain the active polypeptide or spray-dry it.

7. The method for preparing the composite peptide dietary supplement according to claim 6, characterized in that: In the second step, high-activity yeast and Aspergillus oryzae are inoculated in a culture medium at a temperature of 30 to 38° C., cultured at 160 rpm for 12 to 48 hours to prepare a binary fermentation liquid A; Lactobacillus bulgaricus, Streptococcus thermophilus and Lactobacillus acidophilus are inoculated in a culture medium at a temperature of 30 to 42° C., cultured for 5 to 20 hours to prepare a fermentation liquid B; in the third step, sorbitol, arabinose, tea theanine, 2 to 5 times of pure water and 0.1 to 0.2 times of brown sugar are added to the fermentation tank raw pulp based on the weight of the raw pulp obtained in the first step, boiled and kept for 5 minutes, heating was stopped, the temperature was lowered to obtain a diluted mixed pulp for standby use; in the fourth step, the fermentation time is 1 to 2 days.

8. The method for preparing the composite peptide dietary supplement according to claim 6, characterized in that: In the second step, the culture medium of the yeast agent is 1% yeast extract powder, 75-79% water, 3-5% brown sugar and 15-20% potato.

9. A preparation of a composite peptide dietary supplement for improving qi stagnation constitution, characterized in that: The preparation is in the form of tablets, granules or oral liquid.

10. A use of the composite peptide dietary supplement according to claims 1-5, characterized in that: The application refers to the use in preparing food or medicine suitable for people with qi stagnation constitution.

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