Composite peptide dietary supplement for improving phlegm-damp constitution and preparation method, preparation and application thereof

Abstract

The invention provides a composite peptide dietary supplement for improving phlegm-dampness constitution and a preparation method, a preparation and application thereof, and belongs to the technical field of food biology. On the first aspect, the invention provides the composite peptide dietary supplement, and the composite peptide dietary supplement comprises the following raw materials: poria cocos, orange peel, cinnamon, lily, Chinese yam, rhizoma polygonati, mulberry and radix polygonati officinalis; 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 phlegm-dampness constitution.

Description

Technical Field

[0001] This invention relates to the field of food biotechnology. Specifically, it relates to a complex peptide dietary supplement for improving phlegm-dampness 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 throughout their life cycle, manifested in morphological structure, physiological function, and psychological state. It is characterized by individual differences in structure, function, metabolism, and response to external stimuli, as well as susceptibility to certain etiologies and diseases, and a certain tendency in disease progression and outcome. Epidemiological surveys have found that phlegm-dampness constitution is a common imbalanced constitution among the nine categories of constitutions.

[0004] Obesity is a manifestation of metabolic abnormalities in the human body, and it also brings about pathological changes in metabolism. Many people's concerns about obesity stem not only from its appearance but, more importantly, from the various adverse and even life-threatening pathological changes it causes. Increased cholesterol, uric acid, and blood sugar levels put people on the verge of stroke, gout, and diabetes. Obesity can be caused by both acquired factors such as overeating and congenital factors such as constitution. Traditional Chinese medicine refers to this type of constitution, which is prone to metabolic abnormalities, as phlegm-dampness constitution.

[0005] Phlegm-dampness constitution often arises from various pathogenic factors leading to dysfunction of the internal organs' qi transformation, imbalances in the transportation and transformation of qi, blood, and body fluids, resulting in the accumulation of dampness, which transforms into phlegm. This phlegm-dampness accumulates internally, lingering in the internal organs and, in turn, affecting their function. The causes are as follows: 1. Invasion of cold and dampness: Damp climates, wading through water or being caught in rain, or prolonged residence in damp areas allow dampness to invade the body, impairing the spleen and stomach's function, leading to the accumulation of dampness and phlegm, which then accumulates in the lungs. 2. Improper diet: Frequent overeating, excessive consumption of rich, fatty, and sweet foods damages the spleen and stomach, hindering the distribution of nutrients and the transportation and transformation of dampness, resulting in the internal generation of dampness and phlegm. 3. Congenital constitution: A constitution with inherent stomach heat, coupled with excessive consumption of rich, fatty, and sweet foods, impairs the spleen's function, leading to the accumulation of dampness and phlegm. 4. Old age and chronic illness: Weakness of the spleen and stomach, reduced transportation and transformation functions, or kidney yang deficiency, impairing the transformation of qi and the movement of water. 5. Lack of exercise. Prolonged periods of lying down or sitting with little movement can lead to poor blood circulation, sluggish spleen and stomach function, and an inability to metabolize fluids, resulting in the accumulation of dampness and the internal generation of phlegm.

[0006] Individuals with phlegm-dampness constitution have significantly higher levels of cholesterol, triglycerides, very low-density lipoprotein, and blood sugar than those without. They often exhibit obesity, abdominal fullness, chest tightness, excessive phlegm, fatigue, a feeling of heaviness and discomfort, a preference for rich, sweet, and alcoholic foods, a large tongue with a white, greasy coating. The condition is frequently caused by cold-dampness invasion, improper diet, congenital factors, old age, chronic illness, and lack of exercise. Symptoms vary depending on the location of phlegm-dampness retention. They are prone to diabetes, stroke, and chest pain, and have poor adaptability to the rainy season and humid environments. The symptoms of phlegm-dampness constitution are diverse, and the main treatment principle is to dry dampness and resolve phlegm.

[0007] Currently, there are no commercially available health foods made from ingredients such as Poria cocos specifically for people with phlegm-dampness constitution. 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 has the effect of regulating the sub-health state of people with phlegm-dampness constitution.

[0008] Therefore, this invention provides a dietary supplement containing a complex peptide of Poria cocos, tangerine peel, cinnamon, lily bulb, yam, Polygonatum sibiricum, mulberry, and Polygonatum odoratum, as well as its preparation method, formulation, and application. Summary of the Invention

[0009] The purpose of this invention is to provide a compound peptide dietary supplement for improving phlegm-dampness constitution, as well as its preparation method, formulation, and application.

[0010] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:

[0011] In a first aspect, the present invention provides a compound peptide dietary supplement for improving phlegm-dampness constitution. The raw materials of the compound peptide dietary supplement include Poria cocos, tangerine peel, cinnamon, lily bulb, yam, Polygonatum sibiricum, and mulberry.

[0012] Furthermore, the ingredients of this complex peptide dietary supplement also include Solomon's seal.

[0013] Furthermore, the ingredients of this compound peptide dietary supplement, by weight, are: Poria cocos 2-10 parts, tangerine peel 2-10 parts, cinnamon 2-8 parts, lily bulb 2-8 parts, yam 2-8 parts, polygonatum 2-8 parts, mulberry 2-8 parts, and polygonatum odoratum 2-8 parts.

[0014] Furthermore, the ingredients of this compound peptide dietary supplement, by weight, are: 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; or 8 parts Poria cocos, 8 parts tangerine peel, 6 parts cinnamon, 6 parts lily bulb, 6 parts yam, 6 parts Polygonatum sibiricum, 4 parts mulberry, and 4 parts Polygonatum odoratum.

[0015] Secondly, the present invention provides a method for preparing a compound peptide dietary supplement for improving phlegm-dampness constitution, comprising:

[0016] The process includes the following steps: Step 1: Preparation of raw slurry; Step 2: Preparation of yeast starter; Step 3: Preparation of diluted mixed slurry; Step 4: Preparation of fermentation broth; Step 5: Yeast autolysis and ultrasonic cell disruption; Step 6: Enzymatic hydrolysis; Step 7: Ultrafiltration; Step 8: Purification; Step 9: Drying.

[0017] Furthermore, the specific process for preparing the raw pulp in the first step is to take raw materials such as Poria cocos, tangerine peel, cinnamon, lily bulb, yam, Polygonatum sibiricum, mulberry, and Polygonatum odoratum, clean them, soak them until there is no dry core, drain the surface water, and crush them into raw pulp for later use.

[0018] Furthermore, the second step of preparing the yeast starter involves selecting heat-resistant, highly active yeast and Aspergillus oryzae for inoculation and mixed culture to produce a binary yeast starter solution A, and then culturing Streptococcus thermophilus to produce yeast starter solution B, which is ready for use.

[0019] Furthermore, the specific process for preparing the diluted mixed slurry in the third step is as follows: using the weight of the original slurry obtained in the first step as the standard, add an appropriate amount of purified water and an appropriate amount of brown sugar 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.

[0020] Furthermore, the fourth 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.

[0021] Furthermore, the fifth 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.

[0022] Furthermore, the specific process of the sixth enzymatic hydrolysis step is as follows: using the weight of the original slurry obtained in the first step as a standard, add 15u / g to 25u / g of pepsin and plant protease respectively, adjust the pH to 2 to 4, the temperature to 32℃ to 42℃, and the time to 6h to 8h; the enzyme inactivation temperature is 90℃ to 100℃, and the time is 25min to 35min.

[0023] Furthermore, the specific process of the seventh step of 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.

[0024] Furthermore, the eighth step of purification involves using the anion exchanger DEAE-Sephadex A-25 for purification.

[0025] Furthermore, the ninth step, drying, involves freeze-drying the purified product to obtain active peptides or spray-drying it.

[0026] Furthermore, in the second step, highly active yeast and Aspergillus oryzae are inoculated into a culture medium at a temperature of 30–38°C and cultured at 160 rpm for 12–48 hours to prepare binary yeast culture A; Streptococcus thermophilus is inoculated into a culture medium at a temperature of 39–45°C and cultured for 5–20 hours to prepare yeast culture B.

[0027] 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 brown sugar 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.

[0028] Furthermore, in the fourth step, the fermentation temperature is 30–36°C, and the time is 24–48 hours.

[0029] Furthermore, in the second step, the culture medium consists of 1% yeast extract, 75-79% water, 3-5% brown sugar, and 15-20% potato.

[0030] Thirdly, the present invention provides a formulation of the compound peptide dietary supplement, which may be a tablet, granule or oral liquid.

[0031] 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 phlegm-dampness constitution.

[0032] Compared with the prior art, the beneficial effects of the present invention include:

[0033] 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 effects in improving phlegm-dampness constitution. It has significant advantages in many aspects. Detailed Implementation

[0034] 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.

[0035] The features and performance of the present invention will be further described in detail below with reference to embodiments:

[0036] Example 1

[0037] 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, and 6 parts mulberry. Wash them thoroughly, soak them until there is no dry core, drain the surface water, and crush them into raw pulp for later use.

[0038] 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 at 2.5% inoculation rates, and cultured together at 160 rpm for 30 hours at 34°C to prepare binary yeast starter solution A. Activated *Streptococcus thermophilus* was inoculated at 2% inoculation rates and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0039] 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.

[0040] Step 4: Prepare 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.

[0041] 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.

[0042] 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.

[0043] 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.

[0044] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0045] Step 9: Drying: The purified product is spray-dried.

[0046] Example 2

[0047] Step 1: Prepare the raw pulp: Take 2 parts Poria cocos, 10 parts tangerine peel, 2 parts cinnamon, 8 parts lily bulb, 2 parts yam, 8 parts Polygonatum sibiricum, 2 parts mulberry, and 8 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.

[0048] The second step is to prepare the yeast starter: the culture medium consists of 1% yeast extract, 79% water, 3% brown sugar, and 17% potato. Activated, heat-resistant, highly active yeast and Aspergillus oryzae are inoculated at a 1:1 ratio, with an inoculation amount of 2% each. The mixture is cultured at 200 rpm at 30°C for 48 hours to prepare binary yeast starter solution A. Activated Streptococcus thermophilus is inoculated at 1% and cultured at 42°C for 5 hours to prepare yeast starter solution B, which is ready for use.

[0049] 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 5 times the amount of purified water and 0.1 times the amount of brown sugar to the fermentation tank, boil and maintain for 7 minutes, then stop heating and cool down to obtain the diluted mixed slurry, which is ready for use.

[0050] 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 0.03 times their weight. The fermentation temperature is 36℃ and the time is 24h.

[0051] Step 5: Yeast autolysis and ultrasonic cell disruption: After fermentation is terminated, the yeast in the fermentation broth is autolyzed under conditions of 57°C for 12 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 1.5 minutes, followed by a 2-minute interval, for a total of 10 cycles.

[0052] Step 6: Enzymatic hydrolysis: Using the weight of the original slurry obtained in Step 1 as a standard, add 15 u / g of pepsin and 25 u / g of plant protease respectively, adjust the pH to 2, set the temperature to 42℃, and the time to 6 h; the enzyme inactivation temperature is 100℃ and the time is 25 min.

[0053] 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.

[0054] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0055] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0056] Example 3

[0057] Step 1: Prepare the raw pulp: Take 10 parts Poria cocos, 2 parts tangerine peel, 8 parts cinnamon, 2 parts lily bulb, 8 parts yam, 2 parts Polygonatum sibiricum, 8 parts mulberry, and 2 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.

[0058] The second step is to prepare the yeast starter: the culture medium consists of 1% yeast extract, 75% water, 5% brown sugar, and 19% potato. Activated, heat-resistant, highly active yeast and Aspergillus oryzae are inoculated at a 1:1 ratio, with an inoculation amount of 3% each. The mixture is cultured at 120 rpm at 38°C for 12 hours to prepare binary yeast starter solution A. Activated Streptococcus thermophilus is inoculated at 3% and cultured at 30°C for 20 hours to prepare yeast starter solution B, which is ready for use.

[0059] 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 2 times the amount of purified water and 0.2 times the amount of brown sugar to the fermentation tank, boil and maintain for 3 minutes, then stop heating and cool down to obtain the diluted mixed slurry, which is ready for use.

[0060] 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 0.05 times their weight. The fermentation temperature is 30℃ and the time is 48h.

[0061] Step 5: Yeast autolysis and ultrasonic cell disruption: After fermentation is terminated, the yeast in the fermentation broth is autolyzed under conditions of 53℃ for 36 hours. Then, ultrasonic cell disruption is performed, with ultrasonic disruption for 0.5 minutes followed by a 0.5-minute interval, for a total of 30 cycles.

[0062] Step 6: Enzymatic hydrolysis: Using the weight of the original slurry obtained in Step 1 as a standard, add 25 u / g of pepsin and 15 u / g of plant protease respectively, adjust the pH to 4, set the temperature to 32℃, and the time to 8 h; the enzyme inactivation temperature is 90℃ and the time is 35 min.

[0063] 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.

[0064] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0065] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0066] Example 4

[0067] 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, 4 parts mulberry, and 4 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.

[0068] 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, ready for use.

[0069] 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.

[0070] Step 4: Prepare 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.

[0071] 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.

[0072] 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.

[0073] 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.

[0074] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0075] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0076] Example 5

[0077] 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.

[0078] 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 at 2.5% inoculation rates, and cultured together at 160 rpm for 30 hours at 34°C to prepare binary yeast starter solution A. Activated *Streptococcus thermophilus* was inoculated at 2% inoculation rates and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0079] 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.

[0080] Step 4: Prepare 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.

[0081] 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.

[0082] 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.

[0083] 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.

[0084] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0085] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0086] Comparative Example 1

[0087] Step 1: Preparation of ultrafine powder: Take 8 parts of Poria cocos, 8 parts of tangerine peel, 6 parts of cinnamon, 6 parts of lily bulb, 6 parts of yam, 6 parts of Polygonatum sibiricum, 6 parts of mulberry, and 6 parts of Polygonatum odoratum. Crush them into powder with a particle size of about 100μm and set aside.

[0088] 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.

[0089] The third step is to obtain the product: After appropriate concentration of the filtrate, the product is obtained by freeze drying.

[0090] Comparative Example 2

[0091] Step 1: Preparation of ultrafine powder: Take 8 parts of Poria cocos, 8 parts of tangerine peel, 6 parts of cinnamon, 6 parts of lily bulb, 6 parts of yam, 6 parts of Polygonatum sibiricum, 6 parts of mulberry, and 6 parts of Polygonatum odoratum. Crush them into powder with a particle size of about 100μm and set aside.

[0092] 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.

[0093] The third step is to obtain the product: After appropriate concentration of the filtrate, the product is obtained by freeze drying.

[0094] Comparative Example 3

[0095] Step 1: Preparation of ultrafine powder: Take 8 parts of Poria cocos, 8 parts of tangerine peel, 6 parts of cinnamon, 6 parts of lily bulb, 6 parts of yam, 6 parts of Polygonatum sibiricum, 6 parts of mulberry, and 6 parts of Polygonatum odoratum. Crush them into powder with a particle size of about 100μm and set aside.

[0096] 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.

[0097] The third step is to obtain the product: After appropriate concentration of the filtrate, the product is obtained by freeze drying.

[0098] Comparative Example 4

[0099] 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.

[0100] 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 inoculation amounts of 2.5% each. The mixture was cultured at 160 rpm at 34°C for 30 hours to prepare binary yeast starter solution A. Activated *Lactobacillus acidophilus* was inoculated at 2% and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0101] 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.

[0102] Step 4: Prepare 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.

[0103] 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.

[0104] 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.

[0105] 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.

[0106] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0107] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0108] Comparative Example 5

[0109] 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.

[0110] 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 at 34°C for 30 hours to prepare binary yeast starter solution A. Activated *Lactobacillus bulgaricus*, *Streptococcus thermophilus*, and *Lactobacillus acidophilus* were inoculated at 2% each, and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0111] 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.

[0112] Step 4: Prepare 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.

[0113] 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.

[0114] 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.

[0115] 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.

[0116] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0117] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0118] Comparative Example 6

[0119] 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.

[0120] 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 at 2.5% inoculation rates, and cultured together at 160 rpm for 30 hours at 34°C to prepare binary yeast starter solution A. Activated *Streptococcus thermophilus* was inoculated at 2% inoculation rates and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0121] 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 glucose 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.

[0122] Step 4: Prepare 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.

[0123] 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.

[0124] 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.

[0125] 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.

[0126] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0127] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0128] Comparative Example 7

[0129] 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.

[0130] 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 at 2.5% inoculation rates, and cultured together at 160 rpm for 30 hours at 34°C to prepare binary yeast starter solution A. Activated *Streptococcus thermophilus* was inoculated at 2% inoculation rates and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0131] 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.

[0132] Step 4: Prepare 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.

[0133] Step 5: Ultrasonic cell disruption: After terminating fermentation, the fermentation broth is subjected to ultrasonic cell disruption, with ultrasonic disruption for 1 minute, followed by a 1-minute interval, for a total of 20 cycles.

[0134] 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.

[0135] 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.

[0136] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0137] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0138] Comparative Example 8

[0139] 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.

[0140] 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 at 2.5% inoculation rates, and cultured together at 160 rpm for 30 hours at 34°C to prepare binary yeast starter solution A. Activated *Streptococcus thermophilus* was inoculated at 2% inoculation rates and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0141] 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.

[0142] Step 4: Prepare 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.

[0143] Step 5: Yeast autolysis: After fermentation is terminated, the yeast in the fermentation broth is autolyzed. The conditions for yeast autolysis are controlled at 55℃ for 24 hours.

[0144] 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.

[0145] 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.

[0146] Step 8: Purification: Purify using the anion exchanger DEAE-Sephadex A-25.

[0147] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0148] Comparative Example 9

[0149] 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.

[0150] 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 at 2.5% inoculation rates, and cultured together at 160 rpm for 30 hours at 34°C to prepare binary yeast starter solution A. Activated *Streptococcus thermophilus* was inoculated at 2% inoculation rates and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0151] 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.

[0152] Step 4: Prepare 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.

[0153] 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.

[0154] 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.

[0155] Step 7: Purification: Use the anion exchanger DEAE-Sephadex A-25 for purification.

[0156] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0157] Comparative Example 10

[0158] 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.

[0159] 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 at 2.5% inoculation rates, and cultured together at 160 rpm for 30 hours at 34°C to prepare binary yeast starter solution A. Activated *Streptococcus thermophilus* was inoculated at 2% inoculation rates and cultured at 42°C for 12 hours to prepare yeast starter solution B, ready for use.

[0160] 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.

[0161] Step 4: Prepare 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.

[0162] 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.

[0163] 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.

[0164] 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.

[0165] Step 8: Purification: Purify using 724 cation exchange resin.

[0166] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0167] Comparative Example 11

[0168] 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.

[0169] The second step of enzymatic hydrolysis: Using the weight of the original slurry obtained in the first step as a standard, 20 u / g of pepsin and plant protease were added respectively, the pH was adjusted to 3, the temperature was 37℃, and the time was 7 h; the enzyme inactivation temperature was 95℃, and the time was 30 min.

[0170] The third 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.

[0171] Step 4: Purification: Use the anion exchanger DEAE-Sephadex A-25 for purification.

[0172] Step 5: Drying: The purified product is freeze-dried to obtain active peptides.

[0173] Comparative Example 12

[0174] 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.

[0175] 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 *Hepialus hepiali* spores are 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, which is ready for use.

[0176] 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.

[0177] 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.

[0178] 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.

[0179] 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.

[0180] Step 7: Purification: Purify using 724 cation exchange resin.

[0181] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0182] Comparative Example 13

[0183] 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.

[0184] 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.

[0185] 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.

[0186] 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.

[0187] 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.

[0188] 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.

[0189] Step 7: Purification: Purification was performed using Tehua I RA204 anion exchange resin.

[0190] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0191] Comparative Example 14

[0192] 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.

[0193] 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.

[0194] 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.

[0195] Step 4: Prepare 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.

[0196] 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.

[0197] 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.

[0198] Step 7: Purification: Purify using 732 type cation exchange resin.

[0199] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0200] Comparative Example 15

[0201] 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.

[0202] 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.

[0203] 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.

[0204] Step 4: Prepare 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.

[0205] 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.

[0206] 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.

[0207] Step 7: Purification: Purify using TEHUAMP-600 anion exchange resin.

[0208] Step 8: Drying: The purified product is freeze-dried to obtain active peptides.

[0209] Comparative Example 16

[0210] 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.

[0211] 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.

[0212] 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.

[0213] Step 4: Prepare 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.

[0214] 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.

[0215] 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.

[0216] 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.

[0217] Step 8: Purification: Purify using dextran gel G-50.

[0218] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0219] Comparative Example 17

[0220] 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.

[0221] The second step is to prepare the yeast starter: The culture medium consists of yeast extract 10.0 g / L, peptone 20.0 g / L, glycerol 10.0 mL / L, 100.0 mL / L phosphate buffer, magnesium sulfate 0.2 g / L, riboflavin 0.2 mg / L, and histidine 0.2 mg / L. Activated, heat-resistant, and highly active yeast is selected, with an inoculation amount of 2.5%, cultured at 160 rpm and 39°C with aeration for 30 h 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.

[0222] 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.

[0223] 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.

[0224] 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.

[0225] 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.

[0226] 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.

[0227] Step 8: Purification: Purify using dextran gel G-25.

[0228] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0229] Comparative Example 18

[0230] 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.

[0231] 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.

[0232] 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 2.5% of its original volume, 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 original volume, cultured at 36°C for 12 hours to prepare yeast starter solution B, which is ready for use.

[0233] 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.

[0234] 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.

[0235] Step 6: Yeast autolysis and ultrasonic cell disruption: After terminating fermentation, the yeast in the fermentation broth is autolyzed under conditions of 55℃ 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 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.

[0237] Step 8: Purification: Purify using dextran gel LH-60.

[0238] Step 9: Drying: The purified product is freeze-dried to obtain active peptides.

[0239] Experimental Example 1

[0240] The results are shown in Table 1:

[0241]

[0242]

[0243] Experiment Example 2

[0244] 1. Method

[0245] Healthy male SD rats, 6-8 weeks old and weighing 180-200g, were divided into a normal group, a model group, Examples 1-5, and a comparative group, with 10 rats in each group. The rats were housed at room temperature (20-26℃) and humidity (60%-65%), with free access to food and water. During the experiment, the normal group was fed a standard diet; the model group, Examples 1-5, and the comparative group were fed a high-fat diet and underwent a multi-factor intervention involving dietary indiscretion, overexertion, and exposure to cold. Specifically, a specially designed rat cage was used, with a metal mesh partition in the middle. The rats stood on the partition, with 3cm of water below. Each day, the rats swam in 30cm deep, 20℃ water for 30 minutes, and were also administered 20mL / kg of 4℃ cold water via gavage. During the multi-factor intervention, the daily gavage dosage for Examples 1-5 and the comparative group was calculated using the standard animal equivalent dose conversion factor method (based on 50g of raw medicinal material / kg body weight / day for rats). Rats in the normal group and the model group were given the same weight of starch as those in the group in Example 5, twice daily for 28 consecutive days. Total cholesterol (TC), triglycerides (TG), and high-density lipoprotein (HDL-C) were measured on day 29.

[0246] High-fat feed formula: 50% ordinary feed, 13.3% lard, 15.0% sucrose, 20.0% protein, 1.5% cholesterol, and 0.2% sodium cholate.

[0247] 2. Results

[0248] The statistical results (x±s) of total cholesterol, triglycerides and high-density lipoprotein in each group are shown in Table 2.

[0249]

[0250]

[0251] 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 phlegm-dampness constitution, characterized in that, The compound peptide dietary supplement includes the following ingredients: Poria cocos, tangerine peel, cinnamon, lily bulb, yam, polygonatum, and mulberry.

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

3. The compound peptide dietary supplement according to claim 2, characterized in that, By weight, the following components are present: Poria cocos 2-10 parts, Citrus peel 2-10 parts, Cinnamon 2-8 parts, Lily bulb 2-8 parts, Dioscorea opposita 2-8 parts, Polygonatum sibiricum 2-8 parts, Mulberry 2-8 parts, and Polygonatum odoratum 2-8 parts.

4. The compound peptide dietary supplement according to claim 2, characterized in that, By weight, the following proportions are used: 8 parts Poria cocos, 8 parts Citrus peel, 6 parts Cinnamon, 6 parts Lily bulb, 6 parts Dioscorea opposita, 6 parts Polygonatum sibiricum, 6 parts Mulberry, and 6 parts Polygonatum odoratum; or 8 parts Poria cocos, 8 parts Citrus peel, 6 parts Cinnamon, 6 parts Lily bulb, 6 parts Dioscorea opposita, 6 parts Polygonatum sibiricum, 4 parts Mulberry, and 4 parts Polygonatum odoratum.

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: Prepare the yeast starter; Step 3: Prepare the diluted mixed slurry; Step 4: Prepare the fermentation broth; Step 5: Yeast autolysis and ultrasonic cell disruption; Step 6: Enzymatic hydrolysis; 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 to take raw materials such as Poria cocos, tangerine peel, cinnamon, lily bulb, yam, Polygonatum sibiricum, mulberry, and Polygonatum odoratum, 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 second step involves preparing the yeast starter by selecting heat-resistant, highly active yeast and Aspergillus oryzae for inoculation and mixed culture to produce a binary yeast starter solution A, and then culturing Streptococcus thermophilus to produce yeast starter solution B, which is ready for use. The specific process for preparing the diluted mixed slurry in the third 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 brown sugar 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 specific process for preparing the fermentation broth in the fourth step is to inoculate 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 by weight. The specific process of the fifth 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 sixth enzymatic hydrolysis step is as follows: using the weight of the original slurry obtained in the first step as a standard, add 15u / g to 25u / g of pepsin and plant protease respectively, adjust the pH to 2 to 4, the temperature to 32℃ to 42℃, and the time to 6h to 8h; the enzyme inactivation temperature is 90℃ to 100℃, and the time is 25min to 35min. 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 process for the eighth purification step is purification using the anion exchanger DEAE-Sephadex A-25. 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, In the second step, highly active yeast and Aspergillus oryzae are inoculated into a culture medium at a temperature of 30–38°C and cultured at 160 rpm for 12–48 hours to prepare binary yeast culture A; Streptococcus thermophilus is inoculated into a culture medium at a temperature of 39–45°C and cultured for 5–20 hours to prepare yeast culture B; in the third step, based on the weight of the original slurry obtained in the first step, 2–5 times the amount of purified water and 0.1–0.2 times the amount of brown sugar are added to the original slurry, boiled and kept for 5 minutes, then heating is stopped and the temperature is lowered to obtain a diluted mixed slurry for later use; in the fourth step, the fermentation temperature is 30–36°C and the 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 step, the culture medium for the yeast starter is 1% yeast extract, 75-79% water, 3-5% brown sugar and 15-20% potato.

9. A formulation of a compound peptide dietary supplement for improving phlegm-dampness 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 phlegm-dampness constitution.

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