A method for preparing a complex pentapeptide with lipid-lowering and glucose-lowering functions by using enzymolysis-fermentation-enzymolysis technology and application thereof

Abstract

The present application belongs to the field of biotechnology, and particularly relates to a method for preparing a compound pentapeptide with lipid-lowering and glucose-lowering functions by using enzymolysis-fermentation-enzymolysis technology and application thereof. The present application provides a compound pentapeptide with lipid-lowering and glucose-lowering functions, which is obtained by specifically combining soybean, highland barley, mulberry leaves, ginseng and tricholoma matsutake, and can synergistically improve the lipid-lowering and glucose-lowering effects. In addition, the present application also provides a scheme for preparing polypeptides by using the triple biotransformation technology of enzymolysis-fermentation-enzymolysis combined process, which is beneficial to decomposing plant macromolecular substances into small molecular nutrient components which are easy to absorb, and simultaneously extracting the five-dimensional plant compound pentapeptide with glucose-lowering and lipid-lowering functions, thereby improving the biological activity and functionality of the compound pentapeptide, and providing high-quality raw materials for the prevention and treatment of diabetes and hyperlipidemia. Therefore, the technical scheme of the present application has the advantages of safety and reliability, good effect and no side effects.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biotechnology, and particularly relates to a method for preparing a compound pentapeptide with lipid-lowering and glucose-lowering functions by using enzymolysis-fermentation-enzymolysis technology and application thereof. BACKGROUND

[0002] With the development of global economy and changes in lifestyle, metabolic diseases represented by hyperlipidemia and hyperglycemia have become major public health problems threatening human health. According to statistics of the World Health Organization (WHO), there are more than 463 million diabetic patients worldwide, and the number is expected to increase to 578 million by 2030. The number of people with dyslipidemia is as high as 1.3 billion, and is showing a trend of youth. Such diseases not only seriously affect the quality of life of patients, but also can cause cardiovascular diseases, kidney damage, nerve damage and a series of complications, bringing heavy burden to society and family.

[0003] At present, the drugs used in clinical treatment of hyperlipidemia and hyperglycemia mainly include statins, fibrates, insulin and oral hypoglycemic drugs. Although these drugs can effectively control the indicators in the short term, long-term use is often accompanied by side effects such as liver and kidney function damage, hypoglycemia, gastrointestinal reactions, etc. For example, statin drugs can cause muscle pain and abnormal liver function; insulin therapy can cause weight gain and hypoglycemia risk. Therefore, developing a safe, effective and less side effect natural lipid-lowering and glucose-lowering product has become a research hotspot in the fields of medicine and food. SUMMARY

[0004] The purpose of the present application is to provide a method for preparing a compound pentapeptide with lipid-lowering and glucose-lowering functions by using enzymolysis-fermentation-enzymolysis technology and application thereof, which has the advantages of safety, effectiveness and small side effects.

[0005] The present application provides a compound pentapeptide with lipid-lowering and glucose-lowering functions, which comprises the following raw materials in mass fraction: 20-30 parts of soybean, 15-25 parts of highland barley, 10-20 parts of mulberry leaf, 5-15 parts of ginseng and 5-15 parts of truffle.

[0006] The present application provides a method for preparing the compound pentapeptide described in the above technical solution by using enzymolysis-fermentation-enzymolysis technology, which comprises the following steps:

[0007] Mixing and soaking soybean powder, highland barley powder, mulberry leaf powder, ginseng powder, truffle powder and water to obtain a mixture;

[0008] Mixing the mixture with cellulase, and then sequentially performing first enzymolysis and first inactivation to obtain an enzymolysis solution;

[0009] Mixing the enzymolysis solution, glucose and magnesium sulfate heptahydrate, adjusting the pH of the enzymolysis solution, and then sequentially performing sterilization and cooling to obtain a cooled enzymolysis solution;

[0010] mixing the cooled enzymatic hydrolysate with a composite microbial agent to perform anaerobic fermentation, to obtain a fermentation liquor;

[0011] mixing the fermentation liquor with a composite enzyme, and then sequentially performing second enzymatic hydrolysis, second inactivation, solid-liquid separation and concentration drying, to obtain a composite pentapeptide;

[0012] the composite microbial agent comprises Lactobacillus acidophilus, Lactobacillus plantarum and Bacillus subtilis;

[0013] the composite enzyme comprises neutral protease, alkaline protease and papain.

[0014] Preferably, when mixing and soaking, the mass-volume ratio of the total mass of soybean powder, highland barley powder, mulberry leaf powder, ginseng powder and truffle powder to water is 1g:(2-10)mL;

[0015] the mixing and soaking conditions comprise a temperature of 30-60℃ and a time of 2-12h.

[0016] Preferably, when performing the first enzymatic hydrolysis, the mass ratio of the mixture to cellulase is 100:0.5-4;

[0017] the first enzymatic hydrolysis conditions comprise a temperature of 50-65℃ and a time of 1-4h.

[0018] Preferably, in the composite microbial agent, the mass ratio of Lactobacillus acidophilus, Lactobacillus plantarum and Bacillus subtilis is 1:0.8-1:0.5-0.7;

[0019] the effective viable bacterial count of the Lactobacillus acidophilus, Lactobacillus plantarum and Bacillus subtilis is ≥1.5×10 7 cfu / g, respectively.

[0020] Preferably, when performing the anaerobic fermentation, the volume-mass ratio of the enzymatic hydrolysate to the composite microbial agent is 100mL:5-10g.

[0021] Preferably, when mixing the fermentation liquor with the composite enzyme, the temperature of the fermentation liquor is 45-55℃;

[0022] in the composite enzyme, the mass ratio of neutral protease, alkaline protease and papain is 1-2:0.8-1:1-1.2.

[0023] Preferably, when performing the second enzymatic hydrolysis, the mass ratio of the fermentation liquor to the composite enzyme is 100:2-3;

[0024] the second enzymatic hydrolysis conditions comprise a temperature of 40-60℃ and a time of 4-8h.

[0025] Preferably, the pH value of the enzymatic hydrolysate is adjusted to 6.5-7.5 by potassium dihydrogen phosphate;

[0026] The mass ratio of the enzymatic hydrolysate, glucose and magnesium sulfate heptahydrate is 100:18-20:1-1.5.

[0027] The application provides application of the composite pentapeptide in preparation of a lipid-lowering and / or glucose-lowering product.

[0028] Beneficial effects:

[0029] The application provides a composite pentapeptide with lipid-lowering and glucose-lowering functions, which comprises the following raw materials in mass parts: 20-30 parts of soybean, 15-25 parts of highland barley, 10-20 parts of mulberry leaf, 5-15 parts of ginseng and 5-15 parts of truffle.

[0030] The soybean peptide contains rich branched chain amino acids such as leucine and isoleucine, can promote fat metabolism, and reduce serum cholesterol and triglyceride levels; the beta-glucan in the highland barley peptide has the effects of regulating blood glucose and reducing cholesterol, can form a viscous solution in the intestinal tract, delay the digestion and absorption of carbohydrates, reduce postprandial blood glucose fluctuations, and adsorb bile acids to reduce the reabsorption of cholesterol; the mulberry leaf peptide can not only inhibit the activity of alpha-glucosidase and slow down the decomposition rate of starch and other carbohydrates, but also promote insulin secretion and improve insulin sensitivity, thereby reducing blood glucose levels; the ginseng peptide can enhance the sensitivity of the insulin signaling pathway, promote the transport of glucose transporter 4 (GLUT4) to the cell membrane, accelerate the uptake and utilization of glucose, and inhibit the differentiation and synthesis of fat cells to reduce fat accumulation; the truffle peptide is rich in various amino acids, polysaccharides and trace elements, has the effects of antioxidant, anti-inflammatory and regulation of lipid metabolism. Therefore, the specific combination of the above five kinds of peptides can have a synergistic effect and significantly improve the lipid-lowering and glucose-lowering effect.

[0031] Based on the above technical advantages, the application also provides a method for preparing the composite pentapeptide with lipid-lowering and glucose-lowering functions. Through the first enzymatic hydrolysis, the plant cell wall is effectively broken, the leaching rate and extraction efficiency of effective substances in raw materials such as soybeans, highland barley, mulberry leaves, ginseng and tricholoma matsutake are significantly improved, and the deep utilization of plant raw materials is realized; combined with specific probiotic fermentation, it is beneficial to produce various bioactive substances (such as short-chain fatty acids, vitamins, etc.), enhance the lipid-lowering activity of polypeptides; in addition, the fermentation process can degrade anti-nutritional factors and improve the bioavailability of polypeptides; it is beneficial for the synergistic effect of probiotic metabolites and polypeptides, regulates intestinal flora, and improves metabolic health. Therefore, the application is beneficial to decompose plant macromolecules into small molecular nutrients that are easy to absorb, and at the same time, it is beneficial to extract five-dimensional plant composite pentapeptides with glucose-lowering and lipid-lowering functions, improve the biological activity and functionality of the composite pentapeptide, and provide high-quality raw materials for the prevention and treatment of diabetes and hyperlipidemia. At the same time, the raw materials used in the application are all natural substances, do not contain any chemical synthetic components, are safe and reliable, and have no side effects.

[0032] In addition, compared with the traditional preparation process, the application provides a new method for polypeptide preparation by using the triple bioconversion technology of enzymolysis-fermentation-enzymolysis combined process, which has higher efficiency and product activity, accurately intercepts the target molecular weight interval (180-1500 Da) through ultrafiltration membrane, the product activity fluctuation is ≤5%, the activity retention rate is ≥90% after 6 months of room temperature storage, which is significantly better than the traditional enzymolysis product (usually <70%); in addition, the raw materials such as soybeans and highland barley are widely available, and the fermentation waste residue can be recycled to make organic fertilizer, which meets the green production concept, and the production cost is reduced by 15%-20% compared with similar products. DETAILED DESCRIPTION

[0033] In order to further illustrate the application, the schemes provided by the application are described in detail below in combination with examples, but they cannot be understood as limiting the scope of protection of the application.

[0034] Raw material description: cellulase, neutral protease, alkaline protease, papain, lactobacillus acidophilus, lactobacillus plantarum and bacillus subtilis do not have specific source requirements and can be purchased regularly.

[0035] Example 1 (triple process group)

[0036] A method for preparing a composite pentapeptide (i.e. polypeptide composition) by using enzymolysis-fermentation-enzymolysis technology, the steps are:

[0037] (1) The composite pentapeptide is made from the following raw materials in parts by weight: 25 parts of soybeans, 20 parts of highland barley, 15 parts of mulberry leaves, 10 parts of ginseng and 10 parts of tricholoma matsutake.

[0038] (2) The preparation process is:

[0039] A. Raw material pretreatment: the soybeans are crushed into small pieces, and the highland barley, mulberry leaves, ginseng and tricholoma matsutake are crushed into small piecesGlycine max (Linn.) Merr .), highland barley ( Hordeum vulgare Linn. var. nudum Hook. f. ), mulberry leaves ( Morus alba L. ), Ginseng ( Panax ginsengC. A. Meyer ) and matsutake ( Tricholoma matsutake (Ito et Imai) Singer The following ingredients were washed, dried, and pulverized in sequence, and then passed through an 80-mesh sieve to obtain soybean powder, barley powder, mulberry leaf powder, ginseng powder, and matsutake mushroom powder.

[0040] The soybean powder, barley powder, mulberry leaf powder, ginseng powder and matsutake powder were mixed to obtain a mixed powder; the mixed powder was then mixed with water (deionized water) at a mass ratio of 1:5 and soaked at 40°C for 6 hours to obtain a mixture.

[0041] Add cellulase to the mixture at a mass ratio of 100:2 (i.e., the amount of cellulase added is 2% of the mass of the mixture), and hydrolyze at 55℃ for 4 hours; after hydrolysis, inactivate the enzyme at a high temperature of 90-95℃ for 5 minutes (without filtration) to obtain the hydrolysate.

[0042] B. Compound probiotic fermentation:

[0043] The effective viable bacteria count was 1×10⁻⁶. 8 cfu / g of Lactobacillus acidophilus ( Lactobacillus acidophilus Lactobacillus plantarum ( Lactobacillus plantarum ) and Bacillus subtilis ( Bacillus subtilis Mix them according to a mass ratio of 1:1:0.5 to obtain a compound probiotic agent;

[0044] Add 20g of glucose and 1g of magnesium sulfate heptahydrate to every 100g of enzymatic hydrolysate, and adjust the pH of the enzymatic hydrolysate to 7.0 with potassium dihydrogen phosphate. Sterilize the pH-adjusted enzymatic hydrolysate at 121℃ for 20 minutes. After cooling the sterilized enzymatic hydrolysate to 37℃, inoculate it with compound probiotics.

[0045] Inoculate with compound probiotic agent at an inoculation rate of 8% (v / w) (i.e., the volume-to-mass ratio of sterilized enzymatic hydrolysate to compound probiotics is 100mL:8g); after inoculation, anaerobic fermentation is carried out at 37℃ for 36 hours, with stirring once every 6 hours during the fermentation process at a speed of 120rpm to obtain fermentation broth.

[0046] C. Complex enzymatic hydrolysis:

[0047] Preparation of the complex enzyme: Neutral protease, alkaline protease and papain were mixed in a mass ratio of 2:1:1 to obtain the complex enzyme;

[0048] After the fermentation is completed, the fermentation liquor is warmed to 50°C to obtain a warmed fermentation liquor; a composite enzyme is added to the warmed fermentation liquor (the mass ratio of the composite enzyme to the warmed fermentation liquor is 2:100, that is, the addition amount of the composite enzyme is 2% of the total amount of the warmed fermentation liquor) to perform secondary enzymolysis; during the secondary enzymolysis, 1 mol / L NaOH is added to maintain the pH value of the fermentation liquor at 7.0, the enzymolysis time is 6 hours, and the enzymolysis temperature is 50°C.

[0049] D. Enzyme inactivation: after the above enzymolysis reaction is completed, a reaction liquid is obtained; the reaction liquid is heated to 90°C and maintained for 15 minutes to obtain an enzyme-inactivated reaction liquid.

[0050] E. Separation and purification: the enzyme-inactivated reaction liquid is centrifuged at 4000 r / min for 20 minutes to collect a supernatant; the supernatant is subjected to ultrafiltration through an ultrafiltration membrane with a molecular weight cutoff of 1000 Da to collect a retentate.

[0051] F. Concentration and drying: the ultrafiltration retentate is concentrated under reduced pressure at 60°C to 1 / 3 of the original volume, and then subjected to spray drying (the inlet air temperature is 180°C and the outlet air temperature is 80°C) to obtain the composite pentapeptide.

[0052] Example 2 (traditional enzymolysis group)

[0053] The method for preparing the composite pentapeptide by using the traditional enzymolysis process comprises the following steps:

[0054] (1) The composite pentapeptide is prepared from the following raw materials in parts by weight: 25 parts of soybeans, 20 parts of highland barley, 15 parts of mulberry leaves, 10 parts of ginseng, and 10 parts of truffles.

[0055] (2) The preparation process is as follows:

[0056] A. Raw material pretreatment: the soybeans, highland barley, mulberry leaves, ginseng, and truffles are sequentially cleaned, dried, and crushed, and then passed through an 80-mesh sieve to obtain soybean powder, highland barley powder, mulberry leaf powder, ginseng powder, and truffle powder, respectively;

[0057] The above-mentioned soybean powder, highland barley powder, mulberry leaf powder, ginseng powder, and truffle powder are mixed to obtain a mixed powder; the mixed powder is mixed with water (deionized water) at a mass ratio of 1:10, and then soaked at 30-60°C for 6 hours to obtain a mixture;

[0058] The pH value of the mixture is adjusted to 7.0 by using sodium hydroxide and phosphoric acid to obtain a pH-adjusted mixture;

[0059] B. Composite enzymolysis:

[0060] A complex enzyme (i.e., a mixture of neutral protease, alkaline protease, and papain in a mass ratio of 2:1:1) was added to the pH-adjusted mixture. The amount of the complex enzyme added was 20% of the mixture.

[0061] The combined enzyme was hydrolyzed at 50℃ for 6 hours to obtain the enzymatic hydrolysate;

[0062] C. Enzyme inactivation: The method is the same as D. enzyme inactivation in Example 1;

[0063] D. Separation and purification: The method is the same as E. separation and purification in Example 1.

[0064] E. Concentration and drying: The method is the same as F. Concentration and drying in Example 1.

[0065] Example 3 (Fermentation Process Group)

[0066] The method for preparing complex pentapeptides using fermentation technology includes the following steps:

[0067] (1) The compound pentapeptide is made from the following raw materials in parts by weight: 25 parts soybean peptide, 20 parts highland barley peptide, 15 parts mulberry leaf peptide, 10 parts ginseng peptide and 10 parts matsutake peptide.

[0068] (2) The method is as follows:

[0069] A. Raw material pretreatment: The method is the same as that in Example 1.

[0070] B. Compound probiotic fermentation:

[0071] The effective viable bacteria count was 1×10⁻⁶. 8 cfu / g of Lactobacillus acidophilus ( Lactobacillus acidophilus Lactobacillus plantarum ( Lactobacillus plantarum ) and Bacillus subtilis ( Bacillus subtilis Mix them according to a mass ratio of 1:1:0.5 to obtain a compound probiotic agent;

[0072] Add 20g of glucose and 1g of magnesium sulfate heptahydrate to every 100g of enzymatic hydrolysate, and adjust the pH of the enzymatic hydrolysate to 7.0 with potassium dihydrogen phosphate. Sterilize the pH-adjusted enzymatic hydrolysate at 121℃ for 20 minutes. After cooling the sterilized enzymatic hydrolysate to 37℃, inoculate it with compound probiotics.

[0073] Inoculate with compound probiotic agent at an inoculation rate of 8% (v / w) (i.e., the volume-to-mass ratio of sterilized enzymatic hydrolysate to compound probiotics is 100mL:8g); after inoculation, anaerobic ferment at 30-37℃ for 24-48 hours, stirring once every 6 hours during fermentation at a speed of 100-150rpm to obtain fermentation broth.

[0074] C. Inactivation of enzyme: same as the method of D. Inactivation of enzyme in Example 1;

[0075] D. Separation and purification: same as the method of E. Separation and purification in Example 1.

[0076] E. Concentration and drying: same as the method of F. Concentration and drying in Example 1.

[0077] The polypeptide yield (g / 100g) and reducing sugar content (mg / mL) of the composite pentapeptide in Examples 1-3 were determined respectively, and the results are shown in Table 1.

[0078] Table 1 Polypeptide yield and reducing sugar content of composite pentapeptide in different treatments

[0079]

[0080] It can be seen from the data in Table 1 that the polypeptide yield of the composite pentapeptide prepared by the technical solution in Example 1 is as high as 56g / 100g (i.e. 56%), and the reducing sugar content is as high as 3.2mg / mL, compared with Examples 2 and 3. It can be seen that the technical solution of the present application is beneficial to the obtaining of polypeptide and reducing sugar.

[0081] Test Example Influence of preparation process on lipid-lowering effect

[0082] In order to verify the influence of the preparation process on the lipid-lowering effect of the composite pentapeptide, the following experiments were conducted:

[0083] Experimental animals: SPF level male SD rats with a body weight of 180-220g were selected.

[0084] Experimental method: The rats were randomly divided into 6 groups, 10 rats in each group:

[0085] The experimental groups are set as follows:

[0086] Among them, the basic feed and high-fat feed are routinely purchased from Research Diets company;

[0087] The composite pentapeptides prepared in Examples 1-3 are diluted with sterile water according to 1g:10mL to obtain the diluted composite pentapeptide;

[0088] Normal control group: fed with basic feed, and physiological saline was administered by gavage (the gavage amount of physiological saline was the same as that of the composite pentapeptide);

[0089] Model control group: fed with high-fat feed, and physiological saline was administered by gavage (the gavage amount of physiological saline was the same as that of the composite pentapeptide);

[0090] Traditional enzyme hydrolysis group: fed with high-fat feed, and the diluted composite pentapeptide of Example 2 was administered by gavage (the gavage amount was 200mg / kg・d);

[0091] Fermentation process group: feed high-fat diet, gavage with diluted compound pentapeptide of Example 3 (gavage amount is 200 mg / kg·d);

[0092] Triple process group: feed high-fat diet, gavage with diluted compound pentapeptide of Example 1 (gavage amount is 200 mg / kg·d)

[0093] Positive control group: feed high-fat diet, gavage with simvastatin (10 mg / kg·d).

[0094] Each treatment group was raised in the same environment, and after 4 weeks of continuous administration, the serum lipid indicators were measured using the kit from Nanjing Quanlong Biotechnology Co., Ltd. (Nanjing Jiancheng) (among them, the triglyceride (TG) determination kit is A110-2-1; the total cholesterol (TC) determination kit is A111-2-1; the high-density lipoprotein cholesterol determination kit is A112-2-1; the animal low-density lipoprotein cholesterol determination kit is A113-2-1), and the results are shown in Table 2 below.

[0095] Table 2 Determination results of serum lipids in each treatment

[0096]

[0097] Note: compared with the normal control group, * P <0.05; compared with the model control group, # P <0.05, ## P <0.01.

[0098] As can be seen from the data in Table 2, the compound pentapeptide (polypeptide composition) prepared by the technical process in Example 1 is significantly better than the traditional enzymatic process in reducing serum total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C), and increasing high-density lipoprotein cholesterol (HDL-C). P <0.05), indicating that the fermentation step can effectively enhance the lipid-lowering activity of polypeptides.

[0099] Effect of preparation process on blood glucose regulation

[0100] In order to verify the effect of preparation process on blood glucose regulation of compound pentapeptide, the following experiments were conducted:

[0101] Experimental animals: SPF grade male SD rats, weighing 180-220 g.

[0102] Experimental method: the rats were randomly divided into 6 groups, 10 in each group:

[0103] The experimental groups are set as follows:

[0104] The basal diet and the high-sugar and high-fat diet were purchased from Research Diets Company;

[0105] The complex pentapeptide prepared in Examples 1-3 was diluted with sterile water at 1 g: 10 mL to obtain the diluted complex pentapeptide;

[0106] Normal control group: fed with basal diet, and intragastrically administered with normal saline (the amount of normal saline administered was the same as that of the complex pentapeptide);

[0107] Model control group: fed with high-sugar and high-fat diet, and intragastrically administered with normal saline (the amount of normal saline administered was the same as that of the complex pentapeptide);

[0108] Traditional enzymatic hydrolysis group: fed with high-sugar and high-fat diet, and intragastrically administered with the diluted complex pentapeptide of Example 2 (the amount of administration was 200 mg / kg·d);

[0109] Fermentation process group: fed with high-sugar and high-fat diet, and intragastrically administered with the diluted complex pentapeptide of Example 3 (the amount of administration was 200 mg / kg·d);

[0110] Triple process group: fed with high-sugar and high-fat diet, and intragastrically administered with the diluted complex pentapeptide of Example 1 (the amount of administration was 200 mg / kg·d)

[0111] Positive control group: fed with high-sugar and high-fat diet, and intragastrically administered with metformin (50 mg / kg·d).

[0112] Each treatment group was raised in the same environment, and after 4 weeks of continuous administration, the fasting blood glucose (FBG), 2-hour postprandial blood glucose (2 hours after the first meal after administration), serum insulin (INS), serum triglyceride (TG), and total cholesterol (TC) of each treatment group were determined using the kit from Nanjing Quanlong Biotechnology Co., Ltd. (Nanjing Jiancheng), and the results are shown in Table 3.

[0113] Table 3 Determination results of blood glucose indicators in each treatment

[0114]

[0115] Note: compared with the normal control group, * P <0.05; compared with the model control group, # P <0.05, ## P <0.01, ### P <0.001.

[0116] As can be seen from the data in Table 3, the complex pentapeptide (polypeptide composition) prepared by the preparation process in Example 1 has a significant effect on reducing blood glucose.

[0117] In conclusion, the technical scheme provided by the present application is beneficial to decompose plant macromolecular substances into small molecular nutrient components easy to absorb, and extract five-dimensional plant complex pentapeptide with the functions of reducing blood sugar and blood fat, improve the biological activity and functionality of the complex pentapeptide, and provide high-quality raw materials for the prevention and treatment of diabetes and hyperlipidemia, which is safe and reliable without side effects.

[0118] The above merely describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing a complex pentapeptide using an enzymatic-fermentation-enzymatic technique, characterized by, It comprises the following steps: Mixing and soaking soybean powder, highland barley powder, mulberry leaf powder, ginseng powder, truffle powder and water to obtain a mixture; Mixing the mixture with cellulase, and then sequentially performing first enzymolysis and first inactivation to obtain an enzymolysis solution; Mixing the enzymolysis solution, glucose and magnesium sulfate heptahydrate, adjusting the pH of the enzymolysis solution, and then sequentially performing sterilization and cooling to obtain a cooled enzymolysis solution; Mixing the cooled enzymolysis solution with a compound microbial agent to perform anaerobic fermentation to obtain a fermentation solution; Mixing the fermentation solution with a compound enzyme, and then sequentially performing second enzymolysis, second inactivation, solid-liquid separation and concentration drying to obtain a compound pentapeptide; The compound microbial agent comprises Lactobacillus acidophilus, Lactobacillus plantarum and Bacillus subtilis; The compound enzyme comprises neutral protease, alkaline protease and papain; The compound pentapeptide comprises the following raw materials in mass fraction: 20-30 parts of soybean, 15-25 parts of highland barley, 10-20 parts of mulberry leaf, 5-15 parts of ginseng and 5-15 parts of truffle.

2. The method of claim 1, wherein, When mixing and soaking, the mass ratio of the total mass of soybean powder, highland barley powder, mulberry leaf powder, ginseng powder and truffle powder to the volume of water is 1g: (2-10) mL; The mixing and soaking conditions comprise a temperature of 30-60℃ and a time of 2-12h.

3. The method of claim 1, wherein, When performing the first enzymolysis, the mass ratio of the mixture to cellulase is 100:0.5-4; The first enzymolysis conditions comprise a temperature of 50-65℃ and a time of 1-4h.

4. The method of claim 1, wherein, In the compound microbial agent, the mass ratio of Lactobacillus acidophilus, Lactobacillus plantarum and Bacillus subtilis is 1:0.8-1:0.5-0.7; The effective viable cell number of the Lactobacillus acidophilus, Lactobacillus plantarum and Bacillus subtilis is ≥1.5×10 7 cfu / g, respectively.

5. The method of claim 1, wherein, When performing the anaerobic fermentation, the volume-mass ratio of the enzymolysis solution to the compound microbial agent is 100mL:5-10g.

6. The method of claim 1, wherein, When mixing the fermentation solution with the compound enzyme, the temperature of the fermentation solution is 45-55℃; In the compound enzyme, the mass ratio of neutral protease, alkaline protease and papain is 1-2:0.8-1:1-1.

2.

7. The method according to claim 1 or 6, characterized in that, When performing the second enzymolysis, the mass ratio of the fermentation solution to the compound enzyme is 100:2-3; The second enzymolysis conditions comprise a temperature of 40-60℃ and a time of 4-8h.

8. The method of claim 1, wherein, The pH of the enzymolysis solution is adjusted to 6.5-7.5 by potassium dihydrogen phosphate; The mass ratio of the enzymolysis solution, glucose and magnesium sulfate heptahydrate is 100:18-20:1-1.

5.

9. The compound pentapeptide obtained by the preparation method in any one of claims 1-8.

10. The use of the compound pentapeptide in claim 9 in the preparation of a lipid-lowering and / or glucose-lowering product.

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