Composite peptide dietary supplement for improving constitution with yin deficiency as well as preparation method, preparation and application of composite peptide dietary supplement

Abstract

The invention provides a composite peptide dietary supplement for improving yin deficiency constitution, and a preparation method, a preparation and application thereof, and belongs to the technical field of food biology. In the first aspect, the invention provides the composite peptide dietary supplement, and the composite peptide dietary supplement comprises the following raw materials: common yam rhizome, cinnamon, poria cocos, smoked plum, fructus amomi, fresh rhizoma phragmitis, lotus seed, lily, radix polygonati officinalis, liquorice and mulberry; 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 yin deficiency constitution.

Description

Technical Field

[0001] This invention relates to the field of food biotechnology. Specifically, it relates to a compound peptide dietary supplement for improving yin deficiency constitution, its preparation method, formulation, and application. Background Technology

[0002] With the progress of human society, world medicine is shifting from disease-oriented medicine to medicine focused on the "person." From health to sub-health and then to disease, the influence of constitution cannot be ignored. Various constitutional imbalances are the intrinsic basis for the occurrence of diseases. Furthermore, it is precisely because of differences in constitution that the occurrence and outcome of diseases vary. Human constitution possesses both stability and variability; through intervention, imbalances in constitution can be improved and adjusted, thereby restoring health. Identifying constitution through diet and improving imbalances can not only adjust sub-health states and prevent their progression to disease, but also achieve a proactive preventative healthcare effect.

[0003] In Traditional Chinese Medicine (TCM), constitution refers to the relatively stable, comprehensive characteristics of an individual formed throughout their life cycle (birth, growth, maturity, and aging), manifesting in morphological structure, physiological function, and psychological state. It is characterized by individual differences in structure, function, metabolism, and responses to external stimuli, susceptibility to certain etiologies and diseases, and a particular tendency in disease progression and outcome. Epidemiological surveys have found that Yin deficiency constitution is a common imbalanced constitution among the nine categories of constitutions.

[0004] Yin deficiency constitution refers to a condition where, due to dysfunction of the internal organs, there is a predisposition to insufficient yin fluid, leading to internal heat. Common symptoms include emaciation, flushed cheeks, hot palms and soles, night sweats, irritability, dry mouth, dry hair and skin, and a dry, red tongue with little or no coating. It is often caused by external invasion of dryness-heat pathogens, excessive consumption of warming and drying foods, excessive worry, immoderate sexual activity, or prolonged illness. Disease tendencies include susceptibility to general weakness, seminal emission, and insomnia; tolerance to winter but not summer, and intolerance to heat, dryness, and heat pathogens. Treatment should focus on nourishing yin fluids and clearing heat. The specific yin deficiency should be diagnosed based on the individual organ, using formulas to nourish the yin fluids of the five organs and clear their deficiency heat. Based on the theory of yin-yang interdependence, a small amount of yang-tonifying herbs may be added.

[0005] Yin deficiency constitution is often caused by external invasion of dryness and heat, excessive consumption of warm and dry foods, excessive worry, immoderate sexual activity, and prolonged illness, which leads to dysfunction of the internal organs, depletion of yin fluid, and internal heat. Symptoms include loss of body moisture and nourishment, dryness due to deficiency heat, and restlessness due to deficiency fire.

[0006] Yin deficiency mainly manifests in five syndromes: 1. Lung Yin Deficiency Syndrome. Symptoms: Dry cough, scanty sticky white sputum, or sputum with blood streaks, dry throat, red tongue with little coating, and thready and rapid pulse. Treatment principle: Nourish yin and moisten the lungs, supplemented with clearing lung heat. 2. Heart Yin Deficiency Syndrome. Symptoms: Insomnia with excessive dreaming, five-center heat (heat in the palms, soles, and chest), palpitations, red tongue with little coating, and thready and rapid pulse. Treatment principle: Nourish heart yin, supplemented with clearing heart heat. 3. Spleen and Stomach Yin Deficiency Syndrome. Symptoms: Loss of appetite, hard stools, dry mouth and lips, or even dry retching, red tongue with little coating, and thready and rapid pulse. Treatment principle: Nourish yin and harmonize the stomach, supplemented with clearing stomach heat. 4. Liver Yin Deficiency Syndrome. Symptoms: Dry eyes, headache, dizziness, blurred vision, red tongue with little coating, and wiry and thready and rapid pulse. Treatment principle: Nourish liver yin, supplemented with clearing liver heat. 5. Kidney Yin Deficiency Syndrome. Symptoms: Lower back pain and weakness in the legs, dizziness and tinnitus, seminal emission and forgetfulness, scanty urine, dry hair and skin, red tongue with little or no coating, and a thready and rapid pulse. Treatment principle: Nourish kidney yin, supplemented with clearing heat from the lower burner.

[0007] Yin deficiency syndrome has various causes, and it first leads to the lesion of a certain organ. Since the five organs are interconnected, it can affect other organs over time, and the symptoms are varied.

[0008] The theory of constitution posits that "body constitution is related to disease," meaning that body constitution is not only an internal factor in the occurrence of disease but also influences its development and outcome.

[0009] Currently, there are no commercially available health foods made from ingredients such as Chinese yam specifically for people with Yin deficiency. Therefore, this invention is formulated based on the "Classification and Determination of Traditional Chinese Medicine Constitutions" standard officially released by the China Association of Traditional Chinese Medicine on April 9, 2009. It uses ingredients that are both food and medicine, and employs scientific processes to produce concentrated small-molecule peptide compressed candies. This candy can effectively regulate the sub-health state of people with Yin deficiency.

[0010] Therefore, this invention provides a dietary supplement containing a compound peptide of Chinese yam, cinnamon, poria cocos, dried plum, amomum villosum, fresh reed rhizome, lotus seed, lily bulb, polygonatum odoratum, licorice, and mulberry, as well as its preparation method, formulation, and application. Summary of the Invention

[0011] The purpose of this invention is to provide a compound peptide dietary supplement for improving Yin deficiency constitution, as well as its preparation method, formulation, and application.

[0012] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:

[0013] In a first aspect, the present invention provides a compound peptide dietary supplement for improving Yin deficiency constitution. The raw materials of the compound peptide dietary supplement include Chinese yam, cinnamon, poria cocos, dried plum, amomum villosum, fresh reed rhizome, lotus seed, lily bulb, Solomon's seal rhizome, and licorice.

[0014] Furthermore, the ingredients of this complex peptide dietary supplement also include mulberry.

[0015] Furthermore, the ingredients of this compound peptide dietary supplement, by weight, are: Chinese yam 2-10 parts, cinnamon 2-10 parts, poria cocos 2-8 parts, dried plum 2-8 parts, amomum villosum 2-8 parts, fresh reed rhizome 2-8 parts, lotus seed 2-8 parts, lily bulb 2-8 parts, polygonatum odoratum 2-8 parts, licorice root 2-8 parts, and mulberry 2-8 parts.

[0016] Furthermore, the ingredients of this compound peptide dietary supplement, by weight, are: 8 parts Chinese yam, 8 parts cinnamon, 6 parts Poria cocos, 6 parts dried plum, 6 parts Amomum villosum, 6 parts fresh reed rhizome, 6 parts lotus seed, 6 parts lily bulb, 6 parts Polygonatum odoratum, 6 parts licorice root, and 6 parts mulberry; or 8 parts Chinese yam, 8 parts cinnamon, 6 parts Poria cocos, 6 parts dried plum, 6 parts Amomum villosum, 6 parts fresh reed rhizome, 6 parts lotus seed, 4 parts lily bulb, 4 parts Polygonatum odoratum, 4 parts licorice root, and 4 parts mulberry.

[0017] Secondly, the present invention provides a method for preparing a compound peptide dietary supplement for improving Yin deficiency constitution, comprising:

[0018] The process includes the following steps: Step 1: Preparation of raw slurry; Step 2: Enzymatic hydrolysis; Step 3: Preparation of yeast starter; Step 4: Preparation of diluted mixed slurry; Step 5: Preparation of fermentation broth; Step 6: Yeast autolysis and ultrasonic cell disruption; Step 7: Ultrafiltration; Step 8: Purification; Step 9: Drying.

[0019] Furthermore, the specific process for preparing the raw pulp in the first step is to take Chinese yam, cinnamon, poria cocos, dried plum, amomum villosum, fresh reed rhizome, lotus seed, lily bulb, Solomon's seal rhizome, licorice root, and mulberry, wash them clean, soak them until there is no dry core, drain the surface water, and crush them into raw pulp for later use.

[0020] Furthermore, the specific process for the second enzymatic hydrolysis step is as follows: using the weight of the original pulp obtained in the first step as a standard, add 1000-3000 u / g of cellulase and 1000-2000 u / g of pectinase respectively, adjust the pH to 4.0-5.0, the temperature to 50-54℃, and the time to 4-6 hours.

[0021] The third step, preparing the yeast starter, involves inoculating 2.0–3.0% of highly active yeast into a culture medium at 30–38°C, incubating at 160 rpm for 12–48 hours to produce binary yeast starter solution A; and inoculating 1.5–2.5% of Staphylococcus xylose into a culture medium at 30–42°C for 9–15 hours to produce yeast starter solution B. Both cultures use peptone at 8.0–100 mg / L. 2.0 g / L, beef powder 8.0–12.0 g / L, yeast powder 1.8–2.2 g / L, glucose 18.0–22.0 g / L, dipotassium hydrogen phosphate 1.8–2.2 g / L, sodium acetate 4.0–6.0 g / L, triammonium citrate 1.8–2.2 g / L, manganese sulfate 0.04–0.06 g / L, biotin 0.3–0.5 mg / L, riboflavin 0.1–0.3 mg / L.

[0022] Furthermore, the fourth step of preparing the diluted mixed slurry involves using the weight of the original slurry obtained in the first step as a standard, adding an appropriate amount of purified water and an appropriate amount of peptone to the fermentation tank, boiling it and maintaining it for a period of time, stopping the heating, cooling it down, and obtaining the diluted mixed slurry for later use.

[0023] Furthermore, the fifth step of preparing the fermentation broth involves inoculating the binary yeast broth A and yeast broth B from the second step into the diluted mixed slurry obtained in the third step at a ratio of 0.02 to 0.06 times their weight.

[0024] Furthermore, the sixth step, yeast autolysis and ultrasonic cell disruption, involves controlling suitable conditions for yeast autolysis after fermentation is terminated, autolyzing the yeast in the fermentation broth, and then using ultrasonic cell disruption extraction to obtain abundant yeast small molecule peptides and amino acids.

[0025] Furthermore, the specific process of the seventh step, ultrafiltration, involves taking the supernatant and performing membrane separation using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0026] Furthermore, the eighth step of purification involves using dextran gel LH-60 for purification.

[0027] Furthermore, the ninth step, drying, involves freeze-drying the purified product to obtain active peptides.

[0028] Furthermore, in the third step, using the weight of the original slurry obtained in the first step as a standard, add 2 to 5 times the amount of purified water and 0.1 to 0.2 times the amount of peptone to the original slurry, boil it, keep it for 5 minutes, stop heating, cool it down, and obtain a diluted mixed slurry for later use.

[0029] Furthermore, in the fifth step, the fermentation temperature is 30–36℃, and the time is 24–48 hours.

[0030] Furthermore, in the second enzymatic hydrolysis step, based on the weight of the original pulp obtained in the first step, 2000 u / g of cellulase and 1500 u / g of pectinase were added respectively, the pH was adjusted to 4.5, the temperature was 52℃, and the time was 5h; the enzyme inactivation temperature was 95℃, and the time was 30min.

[0031] The culture medium for the third step consisted of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast extract, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at 2.5% of the culture medium, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast culture solution A. Activated Staphylococcus xylose was inoculated at 2% of the culture medium, cultured at 36°C for 12 hours to prepare yeast culture solution B, which was then ready for use.

[0032] The fourth step is to prepare the diluted mixed slurry. Using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0033] The fifth step, preparing the fermentation broth, involves inoculating the binary yeast broth A and yeast broth B from the third step into the diluted mixed slurry obtained in the fourth step at a ratio of 0.04 times their weight, with the fermentation temperature at 33℃ and the time at 36 hours.

[0034] Thirdly, the present invention provides a formulation of the compound peptide dietary supplement, which may be a tablet, granule or oral liquid.

[0035] Fourthly, the present invention also provides the use of the compound peptide dietary supplement and its preparation in the preparation of food or medicine suitable for people with yin deficiency.

[0036] Compared with the prior art, the beneficial effects of the present invention include:

[0037] Compared with existing technologies, the compound peptide dietary supplement provided by this invention has a more scientific and reasonable formulation, an efficient and low-cost preparation method, high activity and high yield of active peptides, high bioavailability of the preparation, and significant improvement in many aspects, including Yin deficiency constitution. Detailed Implementation

[0038] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0039] The features and performance of the present invention will be further described in detail below with reference to embodiments:

[0040] Example 1

[0041] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, and 6 parts of licorice root. Wash them thoroughly, soak them until there is no dry core, drain the surface water, and crush them into raw pulp for later use.

[0042] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0043] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0044] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at a rate of 2.5%, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at a rate of 2%, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0045] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0046] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0047] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0048] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0049] Step 8: Purification: Purify using dextran gel LH-60.

[0050] Example 2

[0051] Step 1: Prepare the raw pulp: Take 10 parts of Chinese yam, 2 parts of cinnamon, 2 parts of poria cocos, 8 parts of dried plum, 2 parts of amomum villosum, 8 parts of fresh reed rhizome, 2 parts of lotus seed, 8 parts of lily bulb, 2 parts of polygonatum odoratum, 8 parts of licorice root, and 2 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0052] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 1000 u / g of cellulase and 2000 u / g of pectinase respectively, adjust the pH to 4.0, set the temperature to 54℃, and the time to 4 h; the enzyme inactivation temperature is 95℃ and the time is 30 min.

[0053] The third step involves preparing the yeast starter: the culture medium consists of 12.0 g / L peptone, 8.0 g / L beef meal, 2.2 g / L yeast powder, 18.0 g / L glucose, 2.2 g / L dipotassium hydrogen phosphate, 4.0 g / L sodium acetate, 2.2 g / L triammonium citrate, 0.04 g / L manganese sulfate, 0.5 mg / L biotin, and 0.1 mg / L riboflavin. Activated, highly active yeast was inoculated at 3.0% in a culture medium at 30°C and cultured at 160 rpm for 48 hours to prepare binary yeast starter solution A. Staphylococcus xylose was inoculated at 1.5% in a culture medium at 42°C and cultured for 9 hours to prepare yeast starter solution B, ready for use.

[0054] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 5 times the amount of purified water and 0.1 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating, cool down, and obtain the diluted mixed slurry for later use.

[0055] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.06 times their weight. The fermentation temperature is 30℃ and the fermentation time is 48h.

[0056] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0057] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0058] Step 8: Purification: Purify using dextran gel LH-60.

[0059] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0060] Example 3

[0061] Step 1: Prepare the raw pulp: Take 2 parts of Chinese yam, 10 parts of cinnamon, 8 parts of poria cocos, 2 parts of dried plum, 8 parts of amomum villosum, 2 parts of fresh reed rhizome, 8 parts of lotus seed, 2 parts of lily bulb, 8 parts of polygonatum odoratum, 2 parts of licorice root, and 8 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0062] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 3000 u / g of cellulase and 1000 u / g of pectinase respectively, adjust the pH to 5.0, set the temperature to 50℃, and the time to 6 h; the enzyme inactivation temperature is 95℃ and the time is 30 min.

[0063] The third step is to prepare the yeast starter: The culture medium consists of 8.0 g / L peptone, 12.0 g / L beef meal, 1.8 g / L yeast powder, 22.0 g / L glucose, 1.8 g / L dipotassium hydrogen phosphate, 6.0 g / L sodium acetate, 1.8 g / L triammonium citrate, 0.06 g / L manganese sulfate, 0.3 mg / L biotin, and 0.3 mg / L riboflavin. Activated, highly active yeast was inoculated at a rate of 2.0% in a culture medium at 38°C and cultured at 160 rpm for 12 hours to prepare binary yeast starter solution A. Staphylococcus xylose was inoculated at a rate of 2.5% in a culture medium at 30°C and cultured for 15 hours to prepare yeast starter solution B, which is ready for use.

[0064] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 2 times the amount of purified water and 0.2 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating, cool down, and obtain the diluted mixed slurry for later use.

[0065] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.02 times their weight. The fermentation temperature is 36℃ and the fermentation time is 24h.

[0066] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0067] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0068] Step 8: Purification: Purify using dextran gel LH-60.

[0069] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0070] Example 4

[0071] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 4 parts of lily bulb, 4 parts of polygonatum odoratum, 4 parts of licorice root, and 4 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0072] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0073] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0074] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at a rate of 2.5%, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at a rate of 2%, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0075] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0076] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0077] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0078] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0079] Step 8: Purification: Purify using dextran gel LH-60.

[0080] Example 5

[0081] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0082] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0083] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0084] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at a rate of 2.5%, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at a rate of 2%, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0085] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0086] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0087] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0088] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0089] Step 8: Purification: Purify using dextran gel LH-60.

[0090] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0091] Comparative Example 1

[0092] Step 1: Preparation of ultrafine powder: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Crush them into powder with a particle size of about 100μm and set aside.

[0093] The second step is water extraction: add 10 times the amount of distilled water, soak for 4 hours, then heat to boiling and decoct for 2 hours, filter, add 5 times the amount of distilled water to the residue, heat to boiling and decoct for 1 hour, and filter; mix the two filtrates.

[0094] The third step is to obtain the product: After appropriate concentration of the filtrate, the product is obtained by freeze drying.

[0095] Comparative Example 2

[0096] Step 1: Preparation of ultrafine powder: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Crush them into powder with a particle size of about 100μm and set aside.

[0097] The second step is ethanol extraction: add 10 times the amount of 95% ethanol, soak for 4 hours, heat and reflux for 2 hours, filter, add 5 times the amount of 95% ethanol to the residue, heat and reflux for 2 hours, filter; mix the two filtrates.

[0098] The third step is to obtain the product: After appropriate concentration of the filtrate, the product is obtained by freeze drying.

[0099] Comparative Example 3

[0100] Step 1: Preparation of ultrafine powder: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Crush them into powder with a particle size of about 100μm and set aside.

[0101] The second step involves ethanol extraction followed by water extraction: Add 10 times the volume of 95% ethanol, soak for 4 hours, then heat under reflux for 2 hours and filter. Add 5 times the volume of 95% ethanol to the residue, heat under reflux for 2 hours and filter. Mix the two filtrates. Add 10 times the volume of distilled water to the residue, soak for 4 hours, then heat to boiling and decoct for 2 hours and filter. Add 5 times the volume of distilled water to the residue, heat to boiling and decoct for 1 hour and filter. Mix the two filtrates with the ethanol extraction filtrate.

[0102] The third step is to obtain the product: After appropriate concentration of the filtrate, the product is obtained by freeze drying.

[0103] Comparative Example 4

[0104] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0105] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0106] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, 2000 u / g of hemicellulase and 1500 u / g of pectinase were added respectively, the pH was adjusted to 4.5, the temperature was 52℃, and the time was 5h; the enzyme inactivation temperature was 95℃ and the time was 30min.

[0107] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at a rate of 2.5%, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at a rate of 2%, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0108] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0109] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0110] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0111] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0112] Step 8: Purification: Purify using dextran gel LH-60.

[0113] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0114] Comparative Example 5

[0115] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0116] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0117] The second step involves preparing the yeast starter: the culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at 2.5% of its population and cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at 2% of its population and cultured at 36°C for 12 hours to prepare yeast starter solution B, ready for use.

[0118] The third step is to prepare the diluted mixed slurry: using the weight of the original slurry prepared in the first step as the standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermentation tank, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry, which is ready for use.

[0119] Step 4: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 2 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0120] Step 5: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0121] Step 6: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0122] Step 7: Purification: Purify using dextran gel LH-60.

[0123] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0124] Comparative Example 6

[0125] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0126] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0127] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0128] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, heat-resistant, and highly active yeast was inoculated at 2.5% of the culture medium, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Lactobacillus bulgaricus was inoculated at 2% of the culture medium, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0129] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0130] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0131] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0132] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0133] Step 8: Purification: Purify using dextran gel LH-60.

[0134] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0135] Comparative Example 7

[0136] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0137] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0138] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0139] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at a rate of 2.5%, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at a rate of 2%, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0140] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of brown sugar to the fermentation tank, boil and maintain for 5 minutes, then stop heating, cool down, and obtain the diluted mixed slurry for later use.

[0141] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0142] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0143] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0144] Step 8: Purification: Purify using dextran gel LH-60.

[0145] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0146] Comparative Example 8

[0147] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0148] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0149] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0150] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at a rate of 2.5%, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at a rate of 2%, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0151] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0152] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0153] Step 6: Ultrasonic cell disruption: After fermentation is terminated, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute, followed by a 1-minute interval, for a total of 20 cycles.

[0154] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0155] Step 8: Purification: Purify using dextran gel LH-60.

[0156] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0157] Comparative Example 9

[0158] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0159] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0160] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0161] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at a rate of 2.5%, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at a rate of 2%, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0162] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0163] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0164] Step 6: Yeast autolysis: After terminating fermentation, the yeast in the fermentation broth is autolyzed. The conditions for yeast autolysis are controlled at 55℃ for 24 hours.

[0165] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0166] Step 8: Purification: Purify using dextran gel LH-60.

[0167] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0168] Comparative Example 10

[0169] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0170] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0171] The second step of enzymatic hydrolysis: Using the weight of the original pulp obtained in the first step as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0172] The third step is to prepare the yeast starter: The culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at a rate of 2.5%, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at a rate of 2%, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0173] The fourth step is to prepare the diluted mixed slurry: using the weight of the enzymatic hydrolysate obtained in the second step as a standard, add 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermenter, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry for later use.

[0174] Step 5: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 3 into the diluted mixed slurry obtained in step 4 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0175] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0176] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0177] Step 8: Purification: Purification using TEHUAMP-600 anion exchange resin.

[0178] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0179] Comparative Example 11

[0180] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0181] The second step is ultrafiltration: After adding 3 times the volume of water to the original slurry and stirring thoroughly, the supernatant is coarsely filtered and then separated using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Daltons to remove large molecular impurities and obtain small molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0182] The third step is purification: purification is carried out using a mixed-bed ion exchange resin with an anion-cation resin ratio of 3:2.

[0183] Step 4: Drying: The purified product is freeze-dried to obtain active peptides.

[0184] Comparative Example 12

[0185] Step 1: Prepare the raw pulp: Take 8 parts ginseng, 6 parts wolfberry, 6 parts hawthorn, 6 parts chicken gizzard lining, 6 parts tangerine peel, 8 parts peach kernel, and 6 parts chicory. Wash them thoroughly, soak them until there is no dry core, drain the surface water, and crush them into raw pulp for later use.

[0186] The second step is to prepare the yeast starter: the culture medium consists of 2% yeast extract, 2% glucose, and 5% soybean meal, with the remainder being water. Activated *Paecilomyces hepiali* is inoculated at 2% of the culture medium and cultured aerobically at 160 rpm for 30 hours at 25°C to prepare binary yeast starter solution A. Activated *Bifidobacterium lactis* is inoculated at 2% of the culture medium and cultured in a sealed container at 36°C for 12 hours to prepare yeast starter solution B, ready for use.

[0187] The third step is to prepare the diluted mixed slurry: Based on the weight of the original slurry obtained in the first step, add 3.5 times the amount of purified water and 0.15 times the amount of brown sugar to the fermentation tank, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry, which is ready for use.

[0188] Step 4: Prepare the fermentation broth. Inoculate the yeast broth A and B from step 2 into the diluted mixed slurry obtained in step 3 at a ratio of 0.04 times their weight. Ferment in a sealed container at 33°C for 36 hours.

[0189] Step 5: Ultrasonic cell disruption. After fermentation is terminated, ultrasonic cell disruption is performed for 1 minute, followed by a 1-minute interval, for a total of 20 cycles.

[0190] Step 6: Ultrafiltration: Take the supernatant and use an ultrafiltration membrane with a molecular weight cutoff of 6000 Daltons for membrane separation to remove macromolecular impurities and obtain peptides and other components with a molecular weight cutoff of less than 6000 Daltons.

[0191] Step 7: Purification: Purify using 724 cation exchange resin.

[0192] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0193] Comparative Example 13

[0194] Step 1: Prepare the raw pulp: Take 8 parts ginseng, 6 parts lotus seeds, 6 parts cardamom, 8 parts yam, 6 parts poria cocos, 8 parts white hyacinth bean, 6 parts tangerine peel, 6 parts licorice root, and 6 parts jujube. Wash them thoroughly, soak them until there is no dry core, drain the surface water, and crush them into raw pulp for later use.

[0195] The second step is to prepare the yeast starter: the culture medium consists of 1% yeast extract, 77% water, 4% brown sugar, and 18% soybean meal ultrafine powder. Activated Kluyveromyces macrocarpa was used, with an inoculation rate of 2.5%, fermented at 160 rpm in a sealed environment at 28°C for 8 hours, followed by aeration for 30 hours to prepare yeast starter solution A. Activated Lactobacillus bulgaricus, Bifidobacterium longum, and Lactobacillus acidophilus were used in an inoculation ratio of 1:1:1, with each inoculated at 2%, and cultured together at 36°C for 12 hours to prepare yeast starter solution B, ready for use.

[0196] The third step is to prepare the diluted mixed slurry: Based on the weight of the original slurry obtained in the first step, add 3.5 times the amount of purified water and 0.15 times the amount of brown sugar to the fermentation tank, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry, which is ready for use.

[0197] Step 4: Prepare the fermentation broth. Inoculate yeast broth A and yeast broth B from step 2 into the diluted mixed slurry obtained in step 3 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0198] Step 5: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0199] Step 6: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0200] Step 7: Purification: Purification using Tehua IRA204 anion exchange resin.

[0201] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0202] Comparative Example 14

[0203] Step 1: Prepare the raw pulp: Take 8 parts of Buddha's Hand, 8 parts of Amomum villosum, 8 parts of Citronella foenum-graecum, 6 parts of Poria cocos, 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 peppermint. Wash the Buddha's Hand, Amomum villosum, Citronella foenum-graecum, Poria cocos, malt, Cordyceps militaris, jujube, licorice, ginseng, and peppermint separately, soak them until no dry core remains, drain the surface water, and crush them into a raw pulp for later use.

[0204] The second step is to prepare the yeast starter: the culture medium consists of 1% yeast extract, 77% water, 4% brown sugar, and 18% potato. Activated, heat-resistant, highly active yeast and *Aspergillus oryzae* were inoculated at a 1:1 ratio, with each inoculated at 2.5% of the total. The mixture was cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated *Lactobacillus bulgaricus*, *Streptococcus thermophilus*, and *Lactobacillus acidophilus* were inoculated at a 1:1:1 ratio, with each inoculated at 2% of the total. The mixture was cultured at 36°C for 12 hours to prepare yeast starter solution B, ready for use.

[0205] 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, 3.5 times the amount of purified water, and 0.15 times the amount of brown sugar to the original slurry in the fermentation tank, boil and maintain for 5 minutes, then stop heating, cool down, and obtain the diluted mixed slurry for later use.

[0206] Step 4: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 2 into the diluted mixed slurry obtained in step 3 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0207] Step 5: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0208] Step 6: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0209] Step 7: Purification: Purify using 732 type cation exchange resin.

[0210] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0211] Comparative Example 15

[0212] Step 1: Prepare the raw pulp: Take 8 parts Poria cocos, 8 parts tangerine peel, 6 parts cinnamon, 6 parts lily bulb, 6 parts yam, 6 parts Polygonatum sibiricum, 6 parts mulberry, and 6 parts Polygonatum odoratum. Wash them thoroughly, soak them until there is no dry core, drain the surface water, and crush them into raw pulp for later use.

[0213] The second step is to prepare the yeast starter: the culture medium consists of 1% yeast extract, 77% water, 4% brown sugar, and 18% potato. Activated, heat-resistant, highly active yeast and Aspergillus oryzae are inoculated at a 1:1 ratio, with inoculation amounts of 2.5% each. The mixture is cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Streptococcus thermophilus is inoculated at 2% and cultured at 42°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0214] 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 3.5 times the amount of purified water and 0.15 times the amount of brown sugar to the fermentation tank, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry, which is ready for use.

[0215] Step 4: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 2 into the diluted mixed slurry obtained in step 3 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0216] Step 5: Yeast autolysis and ultrasonic cell disruption: After fermentation is terminated, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0217] Step 6: Enzymatic hydrolysis: Using the weight of the original slurry obtained in Step 1 as a standard, add 20u / g of pepsin and plant protease respectively, adjust the pH to 3, set the temperature to 37℃, and the time to 7h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0218] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0219] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0220] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0221] Comparative Example 16

[0222] Step 1: Preparation of the raw pulp: Take 12 parts sea buckthorn, 8 parts patchouli, 8 parts perilla, 6 parts oats, 6 parts dried plum, 6 parts houttuynia cordata, 6 parts peppermint, 6 parts angelica dahurica, 6 parts fox nut, 2 parts jujube, 2 parts lactitol, 2 parts isomaltooligosaccharide, and 6 parts licorice. Wash the sea buckthorn, patchouli, perilla, oats, dried plum, houttuynia cordata, peppermint, angelica dahurica, fox nut, jujube, and licorice separately, soak until no dry core remains, drain the surface water, and crush into the raw pulp for later use.

[0223] The second step is to prepare the yeast starter: The culture medium consists of 2% yeast extract, 50% water, 9% brown sugar, and 39% potato. Activated, heat-resistant, and highly active yeast is selected, with an inoculation amount of 2%. The culture is carried out at 31°C, 160 rpm, and aerated for 24 hours to prepare yeast starter solution A, which is ready for use. The culture medium consists of 1% yeast extract, 77% water, 4% brown sugar, and 18% potato. Activated Lactobacillus acidophilus is selected, with an inoculation amount of 2%. The culture is carried out at 35°C in a sealed environment for 48 hours to prepare binary yeast starter solution B, which is ready for use.

[0224] The third step is to prepare the diluted mixed slurry: Based on the weight of the original slurry obtained in the first step, add lactitol, isomaltooligosaccharide, 3.5 times the amount of purified water, and 0.15 times the amount of brown sugar to the fermentation tank. After boiling, maintain the temperature for 5 minutes, stop heating, and cool down to obtain the diluted mixed slurry, which is ready for use.

[0225] Step 4: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 2 into the diluted mixed slurry obtained in step 3 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0226] Step 5: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0227] Step 6: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0228] Step 7: Purification: Purify using TEHUAMP-600 anion exchange resin.

[0229] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0230] Comparative Example 17

[0231] Step 1: Prepare the raw pulp: Take 8 parts dandelion, 8 parts Poria cocos, 6 parts Lophatherum gracile, 6 parts white hyacinth bean, 6 parts Coix seed, 6 parts fresh reed rhizome, 6 parts agastache rugosa, 6 parts red adzuki bean, 6 parts honeysuckle, and 6 parts cardamom. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0232] The second step is to prepare the yeast starter: the culture medium consists of 1% yeast extract, 77% water, 4% brown sugar, and 18% potato. Activated, heat-resistant, highly active yeast and *Aspergillus oryzae* were inoculated at a 1:1 ratio, with an inoculation amount of 2.5% each, and cultured at 160 rpm for 30 hours at 34°C to prepare binary yeast starter solution A. Activated *Lactobacillus bulgaricus*, *Kluyveromyces martensii*, and *Lactobacillus acidophilus* were inoculated at a 1:1:1 ratio, with an inoculation amount of 2% each, and cultured at 36°C for 12 hours to prepare yeast starter solution B, ready for use.

[0233] 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 3.5 times the amount of purified water and 0.15 times the amount of brown sugar to the fermentation tank, boil and maintain for 5 minutes, then stop heating, cool down, and obtain the diluted mixed slurry for later use.

[0234] Step 4: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 2 into the diluted mixed slurry obtained in step 3 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0235] Step 5: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0236] Step 6: Enzymatic hydrolysis: Using the weight of the original slurry obtained in Step 1 as a standard, add 25 u / g of Bacillus subtilis protease, adjust the pH to 9.5, set the temperature to 52℃, and the time to 5 h; the enzyme inactivation temperature is 95℃, and the time is 30 min.

[0237] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0238] Step 8: Purification: Purify using dextran gel G-50.

[0239] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0240] Comparative Example 18

[0241] Step 1: Prepare the raw pulp: Take 8 parts cinnamon, 8 parts dried ginger, 6 parts wolfberry, 6 parts raspberry, 6 parts yam, 6 parts polygonatum, 6 parts jujube, and 6 parts licorice. Wash them thoroughly, soak them until there is no dry core, drain the surface water, and crush them into raw pulp for later use.

[0242] The second step is to prepare the yeast starter: The culture medium consists of 10 g / L yeast extract, 20 g / L peptone, 10 mL / L glycerol, 100 mL / L phosphate buffer, 0.2 g / L magnesium sulfate, 0.2 mg / L riboflavin, and 0.2 mg / L histidine. Activated, heat-resistant, and highly active yeast is selected, with an inoculation amount of 2.5%, and cultured at 160 rpm at 39°C for 30 hours with aeration to prepare binary yeast starter A. The culture medium consisted of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast extract, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 1.0 g / L Tween 80, 0.05 g / L manganese sulfate, and 0.4 mg / L biotin. Activated Bifidobacterium adolescentis and Lactobacillus plantarum were used as inoculation at a ratio of 1:1, with an inoculation amount of 2% each. The mixture was cultured together at 36℃ for 12 hours to prepare yeast culture solution B, which was then ready for use.

[0243] 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 3.5 times the amount of purified water, 0.15 times the amount of glucose, and 0.02 times the amount of xylooligosaccharide to the fermentation tank, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry, which is ready for use.

[0244] Step 4: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 2 into the diluted mixed slurry obtained in step 3 at a ratio of 0.04 times their weight. The fermentation temperature is 38℃ and the time is 36h.

[0245] Step 5: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0246] Step 6 Enzymatic hydrolysis: Using the weight of the original slurry obtained in Step 1 as a standard, add 20 u / g of aspartic protease and cysteine ​​protease respectively, adjust the pH to 5.0, set the temperature to 40℃, and the time to 5 h; the enzyme inactivation temperature is 95℃ and the time is 30 min.

[0247] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0248] Step 8: Purification: Purify using dextran gel G-25.

[0249] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0250] Comparative Example 19

[0251] Step 1: Prepare the raw pulp: Take 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry. Wash each ingredient thoroughly, soak until no dry core remains, drain the surface water, and crush into raw pulp for later use.

[0252] The second step involves preparing the yeast starter: the culture medium consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast was inoculated at 2.5% of its population and cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated Staphylococcus xylose was inoculated at 2% of its population and cultured at 36°C for 12 hours to prepare yeast starter solution B, ready for use.

[0253] 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 3.5 times the amount of purified water and 0.15 times the amount of peptone to the fermentation tank, boil and maintain for 5 minutes, then stop heating and cool down to obtain the diluted mixed slurry, which is ready for use.

[0254] Step 4: Prepare the fermentation broth. Inoculate the binary yeast broth A and yeast broth B from step 2 into the diluted mixed slurry obtained in step 3 at a ratio of 0.04 times their weight. The fermentation temperature is 33℃ and the time is 36h.

[0255] Step 5: Yeast autolysis and ultrasonic cell disruption: After fermentation is terminated, the yeast in the fermentation broth is autolyzed under conditions of 55°C for 24 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1 minute followed by a 1-minute interval, for a total of 20 cycles.

[0256] Step 6: Enzymatic hydrolysis: Using the weight of the original pulp obtained in Step 1 as a standard, add 2000 u / g of cellulase and 1500 u / g of pectinase respectively, adjust the pH to 4.5, set the temperature to 52℃, and the time to 5h; the enzyme inactivation temperature is 95℃ and the time is 30min.

[0257] Step 7: Ultrafiltration: 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons.

[0258] Step 8: Purification: Purify using dextran gel LH-60.

[0259] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0260] Experimental Example 1

[0261] Table 1:

[0262]

[0263]

[0264] Experimental Example 2

[0265] 1. Method

[0266] An animal model of Yin deficiency syndrome was established using the triiodothyronine method. Male mice, weighing 18–22 g, were randomly divided into a normal group, a model group, groups 1–5 (Examples 1–5), and a control group, with 10 mice in each group. The daily gavage dosage for groups 1–5 and the control group was calculated using the standard animal equivalent dose conversion factor (based on 120 g of raw medicinal material / kg body weight / day for mice). Mice in the normal and model groups were administered the same weight of starch as in group 5 via gavage, twice daily for 35 consecutive days. Starting on day 26 of administration, triiodothyronine 18 μg / mouse was administered orally for 10 consecutive days to establish the model. At the end of the experiment, the animals were sacrificed, and plasma cAMP was measured using the protein binding method, and liver glycogen was measured using the anthrone method. The results of the effects on plasma cAMP and liver glycogen in Yin deficiency mice are shown in Table 2.

[0267] Table 2

[0268]

[0269]

[0270] Experimental Example 3

[0271] 1. Method

[0272] Rats (half male and half female, weighing 180–220 g) were randomly divided into a normal group, a model group, groups of Examples 1–5, and a control group, with 10 rats in each group. The model group, groups of Examples 1–5, and the control group were subcutaneously injected with 0.3 g / 100 g body weight of 50% carbon tetrachloride peanut oil every 3 days. On the first day of the experiment, rats were orally administered drugs for modeling liver and kidney yin deficiency (reserpine 0.015 mg / 100 g body weight and thyroxine 1 mg / 100 g body weight). The normal group was injected with an equal volume of physiological saline. Simultaneously, the daily gavage dosage for groups of Examples 1–5 and the control group was calculated using the standard animal equivalent dose conversion factor method (based on 80 g of raw medicinal material / kg body weight / day for rats). Rats in the normal and model groups were gavage with the same weight of starch as in group 5, twice daily. After 21 days, serum reduced glutathione levels were measured using conventional methods.

[0273] 2. Results

[0274] Statistical results of serum reduced glutathione assay in each group See Table 3.

[0275] Table 3

[0276]

[0277]

[0278] All aspects not described in detail in this invention are common knowledge that can be readily understood by those skilled in the art. Although the invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which are obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of this invention fall within the scope of protection claimed by this invention.

Claims

1. A compound peptide dietary supplement for improving Yin deficiency constitution, characterized in that, The compound peptide dietary supplement includes the following ingredients: Chinese yam, cinnamon, poria cocos, dried plum, amomum villosum, fresh reed rhizome, lotus seed, lily bulb, polygonatum odoratum, and licorice.

2. The compound peptide dietary supplement according to claim 1, characterized in that, The ingredients of the compound peptide dietary supplement also include mulberry.

3. The compound peptide dietary supplement according to claim 2, characterized in that, By weight, the following components are present: 2-10 parts of Chinese yam, 2-10 parts of cinnamon, 2-8 parts of Poria cocos, 2-8 parts of dried plum, 2-8 parts of Amomum villosum, 2-8 parts of fresh reed rhizome, 2-8 parts of lotus seed, 2-8 parts of lily bulb, 2-8 parts of Polygonatum odoratum, 2-8 parts of licorice root, and 2-8 parts of mulberry fruit.

4. The compound peptide dietary supplement according to claim 2, characterized in that, By weight, the following proportions are used: 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 6 parts of fresh reed rhizome, 6 parts of lotus seed, 6 parts of lily bulb, 6 parts of polygonatum odoratum, 6 parts of licorice root, and 6 parts of mulberry; or 8 parts of Chinese yam, 8 parts of cinnamon, 6 parts of poria cocos, 6 parts of dried plum, 6 parts of amomum villosum, 4 parts of fresh reed rhizome, 4 parts of lotus seed, 4 parts of lily bulb, 4 parts of polygonatum odoratum, 4 parts of licorice root, and 4 parts of mulberry.

5. The method for preparing the compound peptide dietary supplement according to any one of claims 1-4, characterized in that, Includes the following steps: The steps are as follows: Step 1: Prepare the original slurry; Step 2: Enzymatic hydrolysis; Step 3: Prepare the yeast starter; Step 4: Prepare the diluted mixed slurry; Step 5: Prepare the fermentation broth; Step 6: Yeast autolysis and ultrasonic cell disruption; Step 7: Ultrafiltration; Step 8: Purification; Step 9: Drying.

6. The method for preparing the compound peptide dietary supplement according to claim 5, characterized in that: The specific process for preparing the raw slurry in the first step is as follows: take Chinese yam, cinnamon, poria cocos, dried plum, amomum villosum, fresh reed rhizome, lotus seed, lily bulb, Solomon's seal rhizome, licorice root, and mulberry, wash them clean, soak them until there is no dry core, drain the surface water, and crush them into raw slurry for later use. The specific process for the second step of enzymatic hydrolysis is as follows: The specific process for preparing the fermenting agent in the third step is as follows: select heat-resistant, highly active yeast and Aspergillus oryzae for inoculation and mixed culture to prepare binary fermenting agent A, and mix and culture three bacteria, Lactobacillus bulgaricus, Streptococcus thermophilus and Lactobacillus acidophilus, to prepare fermenting agent B, which is ready for use. The specific process for preparing the diluted mixed slurry in the fourth step is as follows: based on the weight of the original slurry obtained in the first step, add an appropriate amount of purified water and an appropriate amount of peptone to the fermentation tank, boil and maintain for a period of time, stop heating, cool down, and obtain the diluted mixed slurry for later use. The fifth step of preparing the fermentation broth involves inoculating the binary yeast broth A and yeast broth B from the second step into the diluted mixed slurry obtained in the third step at a ratio of 0.02 to 0.06 times their weight. The specific process of the sixth step, yeast autolysis and ultrasonic cell disruption, is as follows: after terminating fermentation, control appropriate yeast autolysis conditions, autolyze the yeast in the fermentation broth, and use ultrasonic cell disruption extraction process to obtain abundant yeast small molecule peptides and amino acids. The specific process of the seventh 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 molecule peptides and other components with a molecular weight cutoff of less than 10,000 Daltons. The specific purification process in the eighth step is to use dextran gel LH-60 for purification. The specific process for the ninth step of drying is to freeze-dry the purified product to obtain active peptides or spray-dry it.

7. The method for preparing the compound peptide dietary supplement according to claim 6, characterized in that, The second enzymatic hydrolysis step uses the weight of the original slurry obtained in the first step as a standard, adding 1000-3000 u / g of cellulase and 1000-2000 u / g of pectinase respectively, adjusting the pH to 4.0-5.0, maintaining the temperature at 50-54℃, and culturing for 4-6 hours. In the third step, a high-activity yeast inoculation of 2.0-3.0% is placed in a culture medium at 30-38℃, cultured at 160 rpm for 12-48 hours to prepare binary yeast culture solution A; a Staphylococcus xylose inoculation of 1.5-2.5% is placed in a culture medium at 30-42℃ and cultured for 9-15 hours to prepare yeast culture solution B. Both culture media contain 8.0-12.0 g / L peptone and 8 g / L beef meal. The following ingredients are listed: 0.0–12.0 g / L yeast powder, 1.8–2.2 g / L glucose, 18.0–22.0 g / L dipotassium hydrogen phosphate, 1.8–2.2 g / L sodium acetate, 4.0–6.0 g / L triammonium citrate, 1.8–2.2 g / L manganese sulfate, 0.04–0.06 g / L biotin, 0.3–0.5 mg / L riboflavin, and 0.1–0.3 mg / L riboflavin. In the fourth step, using the weight of the enzymatic hydrolysate obtained in the second step as a standard, 2–5 times the amount of purified water and 0.1–0.2 times the amount of peptone are added to the original slurry. After boiling, the mixture is kept at a constant temperature for 5 minutes, then heating is stopped, and the temperature is lowered to obtain a diluted mixed slurry for later use. In the fifth step, the fermentation temperature is 30–36℃, and the fermentation time is 24–48 hours.

8. The method for preparing the compound peptide dietary supplement according to claim 6, characterized in that, In the second enzymatic hydrolysis step, the weight of the original pulp obtained in the first step was used as the standard. 2000 u / g of cellulase and 1500 u / g of pectinase were added respectively, the pH was adjusted to 4.5, the temperature was 52℃, and the time was 5 h; the enzyme inactivation temperature was 95℃, and the time was 30 min. The culture medium in the third step consists of 10.0 g / L peptone, 10.0 g / L beef meal, 2.0 g / L yeast extract, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 5.0 g / L sodium acetate, 2.0 g / L triammonium citrate, 0.05 g / L manganese sulfate, 0.4 mg / L biotin, and 0.2 mg / L riboflavin. Activated, thermostable, and highly active yeast is inoculated at 2.5% of the culture medium, cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast culture solution A. Activated Staphylococcus xylose is inoculated at 2% of the culture medium, cultured at 36°C for 12 hours to prepare yeast culture solution B, which is ready for use. The fourth step of preparing the diluted mixed slurry is based on the weight of the enzymatic hydrolysate obtained in the second step. 3.5 times the amount of purified water and 0.15 times the amount of peptone are added to the fermenter, boiled and kept for 5 minutes, then heating is stopped and the mixture is cooled to obtain the diluted mixed slurry for later use. The fifth step involves preparing the fermentation broth by inoculating the binary yeast broth A and yeast broth B from the third step into the diluted mixed slurry obtained in the fourth step at a ratio of 0.04 times their respective weights. The fermentation temperature is 33°C, and the time is 36 hours.

9. A formulation of a compound peptide dietary supplement for improving Yin deficiency constitution, characterized in that, The preparation is a tablet, granule, or oral liquid.

10. An application of the compound peptide dietary supplement according to claims 1-5, characterized in that, The application refers to its use in the preparation of food or medicine suitable for people with Yin deficiency constitution.

Images