Small molecule peptide mixture, freeze-dried powder as well as preparation method and application of freeze-dried powder

Abstract

The invention provides a small molecule peptide mixture, freeze-dried powder as well as a preparation method and application of the freeze-dried powder, and belongs to the technical field of functional foods or health-care products. The small molecule peptide mixture comprises the following raw materials for preparation: plant raw materials and plant lactic acid bacteria, the plant raw materials comprise the following raw materials: rosa roxburghii tratt fruits, poria cocos, Chinese wolfberry fruits, walnuts and radix acanthopanacis senticosi; the plant lactic acid bacteria comprise lactobacillus plantarum, lactobacillus casei and lactobacillus brevis. The rosa roxburghii tratt fruit beverage is obtained by reasonably compounding rosa roxburghii tratt fruits, poria cocos, Chinese wolfberry fruits, walnuts and acanthopanax roots, taking compounded plant raw materials as raw materials, fermenting the raw materials through plant lactic acid bacteria and extracting. Compared with small molecule peptides extracted from a single plant raw material, the small molecule peptide mixture disclosed by the invention has better effects in the aspects of regulating body weight and blood sugar and improving memory.

Description

Technical Field

[0001] The present invention belongs to the technical field of functional foods or health products, and in particular relates to a small molecule peptide mixture, a freeze-dried powder, and a preparation method and application thereof. Background Art

[0002] Rosa roxburghii (RR) is the fruit of the deciduous shrub Rosa roxburghii, also known as thorn berry and spring return, and is a functional food. Rosa roxburghii fruit is rich in nutrients, including polyphenols, flavonoids, polysaccharides and other ingredients, and has antioxidant and anti-tumor effects. However, the health benefits and medicinal value of Rosa roxburghii need to be further developed and utilized. Summary of the invention

[0003] The purpose of the present invention is to provide a small molecule peptide mixture, a lyophilized powder, and a preparation method and application thereof. The small molecule peptide mixture of the present invention has good effects of improving memory and regulating blood sugar.

[0004] The invention provides a small molecule peptide mixture, comprising the following raw materials for preparation: plant raw materials and plant lactic acid bacteria; the plant raw materials include the following raw materials in parts by weight: 5-10 parts of roxburghii, 8-12 parts of tuckahoe, 8-12 parts of wolfberry, 10-15 parts of walnuts and 10-15 parts of acanthopanax senticosus; the plant lactic acid bacteria include Lactobacillus plantarum, Lactobacillus casei and Lactobacillus brevis.

[0005] Preferably, the plant raw materials include the following raw materials in parts by mass: 5 parts of roxburghii, 8 parts of poria, 8 parts of wolfberry, 10 parts of walnuts and 10 parts of Acanthopanax senticosus; or, the plant raw materials include the following raw materials in parts by mass: 10 parts of roxburghii, 12 parts of poria, 12 parts of wolfberry, 15 parts of walnuts and 15 parts of Acanthopanax senticosus.

[0006] Preferably, the plant lactobacillus includes plant lactobacillus ATCC8014; the casei includes casei BCC36987; and the brevis includes brevis D14349.

[0007] The present invention also provides a method for preparing the small molecule peptide mixture described in the above scheme, comprising the following steps: mixing the plant raw material and water to obtain a mixed solution; adjusting the pH value of the mixed solution to 5.8-6.2 and then sterilizing to obtain a fermentation raw material; inoculating plant lactic acid bacteria into the fermentation raw material and fermenting to obtain a fermentation product; mixing the fermentation product with cellulase and performing a first enzymatic hydrolysis to obtain a first enzymatic hydrolysis product; mixing the first enzymatic hydrolysis product with pectinase and performing a second enzymatic hydrolysis to obtain a second enzymatic hydrolysis product; mixing the second enzymatic hydrolysis product with papain and performing a third enzymatic hydrolysis to obtain a third enzymatic hydrolysis product; ultrafiltration of the third enzymatic hydrolysis product, collecting the retentate, and obtaining a small molecule peptide mixture; the ultrafiltration cutoff molecular weight is 2000Da.

[0008] Preferably, the effective viable count of the plant lactic acid bacteria is ≥ 1×10 11 CFU / g; the inoculation amount of the plant lactic acid bacteria is 0.5-1g / L.

[0009] Preferably, the temperature of the first enzymolysis is 55°C; the pH of the first enzymolysis is 6; the time of the first enzymolysis is 3 hours; the temperature of the second enzymolysis is 60°C; the pH of the second enzymolysis is 6.5; the time of the second enzymolysis is 3 hours; the temperature of the third enzymolysis is 60°C; the pH of the third enzymolysis is 6.5; and the time of the third enzymolysis is 1.5 hours.

[0010] The present invention also provides a lyophilized powder of a small molecule peptide mixture, comprising a small molecule peptide mixture and a sweetener; the small molecule peptide mixture is the small molecule peptide mixture described in the above scheme or the small molecule peptide mixture prepared by the preparation method.

[0011] The present invention also provides a method for preparing the lyophilized powder described in the above scheme, comprising the following steps: mixing a small molecule peptide mixture, water and a sweetener to obtain a mixed solution; sterilizing the mixed solution to obtain a sterilized solution; and sequentially freezing and vacuum dehydrating the sterilized solution to obtain a lyophilized powder of the small molecule peptide mixture.

[0012] The present invention also provides the use of the small molecule peptide mixture described in the above scheme, the small molecule peptide mixture prepared by the preparation method, the lyophilized powder or the lyophilized powder prepared by the preparation method in lowering blood sugar, assisting in lowering blood sugar or regulating body weight.

[0013] The present invention also provides the use of the small molecule peptide mixture described in the above scheme, the small molecule peptide mixture prepared by the preparation method, the lyophilized powder or the lyophilized powder prepared by the preparation method in improving memory or assisting in improving memory.

[0014] The present invention provides a small molecule peptide mixture, comprising the following raw materials for preparation: plant raw materials and plant lactic acid bacteria; the plant raw materials include the following raw materials: roxburghii, tuckahoe, wolfberry, walnut and acanthopanax senticosus; the plant lactic acid bacteria include plant lactobacillus, casei lactobacillus and brevis lactobacillus. The present invention is obtained by rationally compounding roxburghii with tuckahoe, wolfberry, walnut and acanthopanax senticosus, and using the compounded plant raw materials as raw materials, and fermenting and extracting with plant lactic acid bacteria. Compared with the small molecule peptides extracted from a single plant raw material, the small molecule peptide mixture of the present invention has better efficacy in regulating body weight, blood sugar and improving memory. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0016] Figure 1 This is an example diagram of a mouse maze test;

[0017] Figure 2 Comparison of the time it takes mice to exit the maze after drug administration;

[0018] Figure 3 This is the result diagram of the weight change of mice before and after administration;

[0019] Figure 4 This is a graph showing changes in blood glucose in mice before and after administration. DETAILED DESCRIPTION

[0020] The invention provides a small molecule peptide mixture, comprising the following raw materials for preparation: plant raw materials and plant lactic acid bacteria; the plant raw materials include the following raw materials in parts by weight: 5-10 parts of roxburghii, 8-12 parts of tuckahoe, 8-12 parts of wolfberry, 10-15 parts of walnuts and 10-15 parts of acanthopanax senticosus; the plant lactic acid bacteria include Lactobacillus plantarum, Lactobacillus casei and Lactobacillus brevis.

[0021] In the present invention, the plant raw material is preferably composed of the following raw materials in parts by mass: 5-10 parts of roxburghii, 8-12 parts of tuckahoe, 8-12 parts of wolfberry, 10-15 parts of walnut and 10-15 parts of acanthopanax senticosus.

[0022] In the present invention, the roxburghii, tuckahoe, wolfberry, walnut and acanthopanax senticosus are preferably derived from commercially available dry products.

[0023] In the present invention, the plant raw materials preferably include the following raw materials in parts by mass: 5 parts of roxburghii, 8 parts of poria, 8 parts of wolfberry, 10 parts of walnuts and 10 parts of Acanthopanax senticosus; or, the plant raw materials preferably include the following raw materials in parts by mass: 10 parts of roxburghii, 12 parts of poria, 12 parts of wolfberry, 15 parts of walnuts and 15 parts of Acanthopanax senticosus.

[0024] In one embodiment of the present invention, the plant raw material is composed of the following raw materials in parts by weight: 5 parts of roxburghii, 8 parts of tuckahoe, 8 parts of wolfberry, 10 parts of walnut and 10 parts of Acanthopanax senticosus.

[0025] In another embodiment of the present invention, the plant raw material is composed of the following raw materials in parts by weight: 10 parts of roxburghii, 12 parts of tuckahoe, 12 parts of wolfberry, 15 parts of walnut and 15 parts of Acanthopanax senticosus

[0026] In the present invention, the plant lactic acid bacteria are preferably composed of Lactobacillus plantarum, Lactobacillus casei and Lactobacillus brevis; the ratio of the effective viable counts of Lactobacillus plantarum, Lactobacillus casei and Lactobacillus brevis is preferably 1:1:1. In the present invention, the plant lactobacillus preferably includes Lactobacillus plantarum ATCC8014; the Lactobacillus casei preferably includes Lactobacillus casei BCC36987; the Lactobacillus brevis preferably includes Lactobacillus brevis D14349; in the present invention, the plant lactic acid bacteria are preferably purchased from Shanghai Preservation Biotechnology Center.

[0027] The present invention also provides a method for preparing the small molecule peptide mixture described in the above scheme, comprising the following steps:

[0028] mixing the plant material and water to obtain a mixed solution;

[0029] The pH value of the mixed solution is adjusted to 5.8-6.2 and then sterilized to obtain a fermentation raw material;

[0030] Inoculating plant lactic acid bacteria into the fermentation raw material, performing fermentation, and obtaining a fermentation product;

[0031] The fermentation product is mixed with cellulase and subjected to a first enzymolysis to obtain a first enzymolysis product;

[0032] The first enzymatic hydrolysis product is mixed with pectinase, and subjected to a second enzymatic hydrolysis to obtain a second enzymatic hydrolysis product;

[0033] The second enzymatic hydrolysis product is mixed with papain, and subjected to a third enzymatic hydrolysis to obtain a third enzymatic hydrolysis product;

[0034] The third enzymatic hydrolysis product is subjected to ultrafiltration, and the retentate is collected to obtain a small molecule peptide mixture; the ultrafiltration molecular weight cutoff is 2000Da.

[0035] The present invention firstly mixes the plant raw material and water to obtain a mixed solution.

[0036] Before mixing the plant material and water, the present invention preferably further comprises crushing the plant material to obtain plant material powder.

[0037] In the present invention, the mass ratio of the plant raw material to water is preferably (40-70):350, more preferably (41-64):350. In the present invention, the water is preferably distilled water.

[0038] After obtaining the mixed solution, the present invention adjusts the pH value of the mixed solution to 5.8-6.2 and then sterilizes it to obtain the fermentation raw material.

[0039] In the present invention, the reagent used to adjust the pH value of the mixed solution is preferably baking soda; the pH value of the mixed solution is preferably adjusted to 6. In the present invention, the sterilization temperature is preferably 121° C.; the sterilization time is preferably 20 minutes.

[0040] After obtaining the fermentation raw material, the present invention inoculates plant lactic acid bacteria into the fermentation raw material to perform fermentation to obtain a fermentation product.

[0041] In the present invention, the effective viable count of the plant lactic acid bacteria is preferably ≥ 1×10 11 CFU / g; the inoculation amount of the plant lactic acid bacteria is preferably 0.5-1 g / L. In the present invention, the fermentation temperature is preferably 37°C; the fermentation time is preferably 96h; and the fermentation humidity is preferably 60%.

[0042] After obtaining the fermentation product, the present invention mixes the fermentation product with cellulase and performs a first enzymolysis to obtain a first enzymolysis product.

[0043] In the present invention, the addition amount of the cellulase is preferably 0.1% of the fermentation liquid mass; the enzymatic activity of the cellulase is preferably 100,000 U / g. The temperature of the first enzymatic hydrolysis is preferably 55°C; the pH of the first enzymatic hydrolysis is preferably 6; and the time of the first enzymatic hydrolysis is preferably 3 hours.

[0044] After obtaining the first enzymatic hydrolysis product, the present invention mixes the first enzymatic hydrolysis product with pectinase and performs a second enzymatic hydrolysis to obtain a second enzymatic hydrolysis product.

[0045] In the present invention, the added amount of the pectinase is preferably 0.1% of the fermentation liquid mass; the enzyme activity of the pectinase is preferably 100,000 U / g; the temperature of the second enzymatic hydrolysis is preferably 60°C; the pH of the second enzymatic hydrolysis is preferably 6.5; and the time of the second enzymatic hydrolysis is preferably 3h.

[0046] After obtaining the second enzymatic hydrolysis product, the present invention mixes the second enzymatic hydrolysis product with papain and performs a third enzymatic hydrolysis to obtain a third enzymatic hydrolysis product.

[0047] In the present invention, the addition amount of papain is preferably 0.1% of the fermentation broth mass; the enzymatic activity of papain is preferably 100,000 U / g; the temperature of the third enzymatic hydrolysis is preferably 60°C; the pH of the third enzymatic hydrolysis is preferably 6.5; and the time of the third enzymatic hydrolysis is preferably 1.5h.

[0048] After the third enzymatic hydrolysis, the present invention preferably further comprises enzyme inactivation, and the enzyme inactivation is preferably performed by a high temperature method.

[0049] After obtaining the third enzymatic hydrolysis product, the present invention performs ultrafiltration on the third enzymatic hydrolysis product, collects the retentate, and obtains a small molecule peptide mixture; the ultrafiltration molecular weight cutoff is 2000Da.

[0050] The present invention also provides a lyophilized powder of a small molecule peptide mixture, comprising a small molecule peptide mixture and a sweetener; the small molecule peptide mixture is the small molecule peptide mixture described in the above scheme or the small molecule peptide mixture prepared by the preparation method.

[0051] In the present invention, the mass percentage of the sweetener in the freeze-dried powder is preferably 2% to 5.5%; the sweetener is preferably xylitol.

[0052] The present invention also provides a method for preparing the freeze-dried powder described in the above scheme, comprising the following steps:

[0053] Mixing the small molecule peptide mixture, water and a sweetener to obtain a mixed solution;

[0054] Sterilizing the mixed solution to obtain a sterilized solution;

[0055] The sterilized liquid is sequentially frozen and vacuum-dehydrated to obtain a lyophilized powder of a small molecule peptide mixture.

[0056] The invention firstly mixes a small molecule peptide mixture, water and a sweetener to obtain a mixed solution.

[0057] In the present invention, the mixing of the small molecule peptide mixture, water and sweetener preferably includes first dissolving the small molecule peptide mixture in water and then mixing with the sweetener. In the present invention, the water is preferably ultrapure water.

[0058] After obtaining the mixed solution, the present invention sterilizes the mixed solution to obtain a sterilized solution.

[0059] In the present invention, the sterilization temperature is preferably 120-121° C.; the sterilization time is preferably 15 min.

[0060] After obtaining the sterilization solution, the present invention sequentially freezes and vacuum dehydrates the sterilization solution to obtain a freeze-dried powder of a small molecule peptide mixture.

[0061] In the present invention, the sterilizing liquid is preferably placed in a container for freezing. In the present invention, the freezing temperature is preferably -20°C; the freezing time is based on the freezing of the sterilizing liquid. In the present invention, the role of the vacuum dehydration is to remove water from the frozen sterilizing liquid by evaporation in a vacuum environment, so that the water in the solution evaporates and leaves the freeze-dried active ingredients. In the present invention, the conditions of the vacuum dehydration are preferably: -42°C, 50Pa, 30h; the vacuum freezing is preferably carried out using a vacuum dryer, the machine model is SCIENTZ-10N.

[0062] The present invention also provides the use of the small molecule peptide mixture described in the above scheme, the small molecule peptide mixture prepared by the preparation method, the lyophilized powder or the lyophilized powder prepared by the preparation method in lowering blood sugar, assisting in lowering blood sugar or regulating body weight.

[0063] In the present invention, the regulating body weight preferably includes reducing body weight (losing weight).

[0064] In the present invention, the use in lowering blood sugar, assisting in lowering blood sugar or regulating body weight includes the use in preparing products having the effects of lowering blood sugar, assisting in lowering blood sugar or regulating body weight. In the present invention, the products include food, health products or medicines.

[0065] The present invention also provides the use of the small molecule peptide mixture described in the above scheme, the small molecule peptide mixture prepared by the preparation method, the lyophilized powder or the lyophilized powder prepared by the preparation method in improving memory or assisting in improving memory.

[0066] In the present invention, the use in improving memory or assisting in improving memory includes the use in preparing a product having the effect of improving memory or assisting in improving memory. In the present invention, the product includes food, health care product or medicine.

[0067] In the present invention, the food includes functional food.

[0068] In order to further illustrate the present invention, a small molecule peptide mixture, a lyophilized powder, and a preparation method and application thereof provided by the present invention are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0069] In the embodiment of the present invention, Lactobacillus plantarum is Lactobacillus plantarum ATCC8014, Lactobacillus brevis is Lactobacillus brevis D14349, and Lactobacillus casei is Lactobacillus casei BCC36987, all of which are purchased from Shanghai Preservation Biotechnology Center, and the dosage forms are all freeze-dried powders.

[0070] Example 1

[0071] Ingredients: 5g sea buckthorn, 8g poria, 8g wolfberry, 10g walnut, and 10g Acanthopanax.

[0072] 1) Crush roxburghii, Poria cocos, wolfberry, walnut and Acanthopanax senticosus into powder using a pulverizer to obtain plant raw material powder.

[0073] 2) The plant raw material powder and 350 mL of distilled water were mixed to obtain a mixed solution, the mixed solution was adjusted to pH 6 with baking soda, and then sterilized at 121° C. for 20 min and cooled to obtain a fermentation raw material;

[0074] 3) inoculating plant lactic acid bacteria into the fermentation raw material, fermenting and obtaining a fermentation product; the plant lactic acid bacteria are composed of Lactobacillus plantarum, Lactobacillus casei and Lactobacillus brevis; the inoculation amount of the plant lactic acid bacteria is 0.5 g / L; the fermentation conditions are: culture conditions at 37° C., fermentation hours for 96 hours, and humidity of 60%;

[0075] 4) Performing proteolysis on the fermentation broth: adding 0.1% of the mass of cellulase to the fermentation broth, the enzyme activity is 100,000 U / g, the enzymatic hydrolysis conditions are 55°C, pH 6, and enzymatic hydrolysis time is 3h; then adding 0.1% of the mass of pectinase to the fermentation broth, the enzyme activity of pectinase is 100,000 U / g, the temperature of pectinase enzymatic hydrolysis is 60°C, pH 6.5, and the enzymatic hydrolysis time is 3h. Then add 0.1% of the mass of papain to the fermentation broth, the enzyme activity of papain is 100,000 U / g, the conditions of papain enzymatic hydrolysis are: pH 6.5, temperature 60°C, time is 1.5h, after enzymatic hydrolysis, use high temperature method to inactivate the enzyme to obtain enzymatic hydrolyzate;

[0076] 5) ultrafiltration of the enzymatic hydrolysate using an ultrafiltration membrane with a molecular weight cutoff of 2000Da to obtain small molecule peptides;

[0077] 6) After the small molecule peptide is dissolved in ultrapure water, 2% xylitol is added thereto by weight, and the mixture is sterilized at 120° C. for 15 min to obtain a small molecule peptide liquid;

[0078] 7) Pour the small molecule peptide liquid into a container and then place it in a -20°C low temperature environment to allow the water in the solution to gradually freeze;

[0079] 8) Vacuum dehydration: Place the frozen small molecule peptide liquid in a vacuum environment, remove water by evaporation, evaporate the water in the solution, leave the freeze-dried active ingredients, and obtain a freeze-dried powder of the small molecule peptide mixture; parameters: temperature -42°C, vacuum 50Pa, time 30h; vacuum dryer, machine model: SCIENTZ-10N.

[0080] Comparative Example 1

[0081] Ingredients: 5g roxburghii

[0082] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0083] Comparative Example 2

[0084] Ingredients: Poria 8g

[0085] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0086] Comparative Example 3

[0087] Ingredients: 8g wolfberry

[0088] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0089] Comparative Example 4

[0090] Ingredients: 8g wolfberry

[0091] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0092] Comparative Example 5

[0093] Ingredients: Acanthopanax senticosus 10g

[0094] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0095] Example 2

[0096] Ingredients: 10g sea buckthorn, 12g poria, 12g wolfberry, 15g walnut, and 15g Acanthopanax.

[0097] The rest is the same as in Example 1.

[0098] Comparative Example 6

[0099] Ingredients: 10g roxburghii

[0100] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0101] Comparative Example 7

[0102] Ingredients: Poria 12g

[0103] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0104] Comparative Example 8

[0105] Ingredients: 12g wolfberry

[0106] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0107] Comparative Example 9

[0108] Ingredients: 15g walnut

[0109] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0110] Comparative Example 10

[0111] Ingredients: Acanthopanax senticosus 15g

[0112] In step 2), the amount of distilled water used is 50 mL, and the rest is the same as in Example 1.

[0113] Test Example 1: Maze Test

[0114] 112 Kunming mice were purchased from Mudanjiang Medical College and adapted to feeding for one week, during which they were free to eat and drink. The mice were randomly divided into 14 groups, A1 to A8 and B2 to B7, as shown below:

[0115] A1: blank control group 1;

[0116] A2: small molecule peptide mixture group 1 (complex), lyophilized powder of the small molecule peptide mixture prepared in Example 1;

[0117] A3: 5 g roxburghii group, freeze-dried powder prepared in Comparative Example 1;

[0118] A4: Poria 8g group, freeze-dried powder prepared in Comparative Example 2;

[0119] A5: wolfberry 8g group, freeze-dried powder prepared in Comparative Example 3;

[0120] A6: walnut 10g group, freeze-dried powder prepared in Comparative Example 4;

[0121] A7: Acanthopanax senticosus 10 g group, freeze-dried powder prepared in Comparative Example 5;

[0122] A8: positive control DHA group, DHA purchased from Tong Ren Tang;

[0123] B2: small molecule peptide mixture group 2 (complex), lyophilized powder of the small molecule peptide mixture prepared in Example 2;

[0124] B3: 10 g roxburghii group, freeze-dried powder prepared in Comparative Example 6;

[0125] B4: Poria 12g group, freeze-dried powder prepared in Comparative Example 7;

[0126] B5: wolfberry 12g group, freeze-dried powder prepared in Comparative Example 8;

[0127] B6: walnut 15g group, freeze-dried powder prepared in Comparative Example 9;

[0128] B7: Acanthopanax 15 g group, freeze-dried powder prepared in Comparative Example 10.

[0129] There were 8 mice in each group (initial body weight 23±2.0g, SPF grade). Except for the blank control group, the drug-treated groups were gavaged with the corresponding lyophilized powder 50mg / kg, and the blank control group was gavaged with 1mL / (mouse / time) of normal saline. The DHA group used DNA purchased from Tong Ren Tang, and gavage was performed with 1mg of capsule contents each time. Twice a day, i.e. once in the morning and once in the evening. At the same time, the blank group was given an equal amount of normal saline, and the drug was continuously gavaged for 4 weeks.

[0130] Maze test: The time taken by mice in different groups to walk through the maze was recorded 7 days, 14 days and 28 days after administration. The example of the mouse maze test is shown in the figure below: Figure 1 The detection equipment used is SA220 from Jiangsu Saiangsi Biotechnology Co., Ltd.

[0131] The average time for each group of mice to walk out of the maze is shown in Tables 1 and 2 and Figure 2 It can be seen that the time taken by mice in the sea buckthorn group to get out of the maze was significantly shortened.

[0132] Table 1 The time taken for mice to exit the maze in the low-concentration drug administration groups (A1-A8 groups)

[0133]

[0134] Table 2 The time taken for mice to exit the maze in the high concentration drug administration groups (B2-B7 groups)

[0135]

[0136]

[0137] Test Example 2

[0138] 1. Grouping

[0139] 105 Kunming mice were purchased from Mudanjiang Medical College and adapted to feeding for one week, during which they were free to eat and drink. The mice were randomly divided into 15 groups, C1-C8 groups, D2-D7 groups and a model group, as shown below:

[0140] C1: blank control group 1;

[0141] C2: small molecule peptide mixture group 1 (complex), lyophilized powder of the small molecule peptide mixture prepared in Example 1;

[0142] C3: 5 g roxburghii group, freeze-dried powder prepared in Comparative Example 1;

[0143] C4: Poria 8g group, freeze-dried powder prepared in Comparative Example 2;

[0144] C5: wolfberry 8g group, freeze-dried powder prepared in Comparative Example 3;

[0145] C6: walnut 10g group, freeze-dried powder prepared in Comparative Example 4;

[0146] C7: Acanthopanax senticosus 10 g group, freeze-dried powder prepared in Comparative Example 5;

[0147] C8: positive metformin group, metformin;

[0148] D2: small molecule peptide mixture group 2 (complex), lyophilized powder of the small molecule peptide mixture prepared in Example 2;

[0149] D3: 10 g roxburghii group, freeze-dried powder prepared in Comparative Example 6;

[0150] D4: Poria 12g group, freeze-dried powder prepared in Comparative Example 7;

[0151] D5: wolfberry 12g group, freeze-dried powder prepared in Comparative Example 8;

[0152] D6: walnut 15g group, freeze-dried powder prepared in Comparative Example 9;

[0153] D7: Acanthopanax senticosus 15 g group, freeze-dried powder prepared in Comparative Example 10;

[0154] Each group consisted of 7 mice (initial body weight 23±2.0g, SPF grade). Except for the blank control group, the drug-treated groups were gavaged with the corresponding lyophilized powder at a dose of 50mg / kg, and the blank control group was gavaged with 1mL / (mouse / time) of normal saline. The positive metformin group was gavaged with the contents of the capsule at 50mg / kg each time. Once a day. At the same time, the blank control group and the model group were given an equal amount of normal saline for 6 consecutive weeks.

[0155] 2. Mouse modeling: After one week of adaptive feeding of mice except the blank group, STZ solution with a concentration of 5 mg / ml was intraperitoneally injected every morning, and 35 mg / kg was injected daily according to body weight for 3 consecutive days. The mice drank tap water and were fed with high-fat and high-sugar feed for mice. For the blank group mice, an equal amount of sodium citrate buffer was intraperitoneally injected daily. The mice drank tap water and were fed with regular feed for mice. The drinking and eating conditions were observed every day. The body weight was tested once a day after modeling. The fasting blood glucose value of the mice was tested three days after modeling. When the blood glucose value of the mice reached 11.1 mmol / L, the modeling was successful.

[0156] 3. Body weight test: Weigh the mice every day, record the weighing results, and monitor the weight changes before and after modeling. The results of mouse weight test are shown in Tables 3 to 16 and Figure 3 It can be seen that the body weight of mice in group C2 and group D2 (complex group) decreased significantly.

[0157] Table 3 Body weight of mice in the low-concentration drug administration groups (C1-C8 groups) before drug administration

[0158]

[0159] Table 4 Body weight of mice in the low-concentration drug administration group (C1-C8 groups) after 1 week of drug administration

[0160]

[0161]

[0162] Table 5 Body weight of mice in the low-concentration drug administration group (C1-C8 groups) after 2 weeks of drug administration

[0163]

[0164] Table 6 Body weight of mice in the low-concentration drug administration group (C1-C8 groups) after 3 weeks of drug administration

[0165]

[0166]

[0167] Table 7 Body weight of mice in the low-concentration drug administration group (C1-C8 groups) after 4 weeks of drug administration

[0168]

[0169] Table 8 Body weight of mice in the low-concentration drug administration group (C1-C8 groups) after 5 weeks of drug administration

[0170]

[0171] Table 9 Body weight of mice in the low-concentration drug administration group (C1-C8 groups) after 6 weeks of drug administration

[0172]

[0173]

[0174] Table 10 Body weight of mice in the high concentration administration group (D2-D7 groups) before administration

[0175]

[0176] Table 11 Body weight of mice in the high concentration administration group (D2-D7 groups) after 1 week of administration

[0177]

[0178]

[0179] Table 12 Body weight of mice in the high concentration administration group (D2-D7 groups) after 2 weeks of administration

[0180]

[0181] Table 13 Body weight of mice in the high concentration administration group (D2-D7 groups) after 3 weeks of administration

[0182]

[0183]

[0184] Table 14 Body weight of mice in the high concentration administration group (D2-D7 groups) after 4 weeks of administration

[0185]

[0186] Table 15 Body weight of mice in the high concentration administration group (D2-D7 groups) after 5 weeks of administration

[0187]

[0188] Table 16 Body weight of mice in the high concentration administration group (D2-D7 groups) after 6 weeks of administration

[0189]

[0190] 4. Blood glucose test: Blood was collected from the tail vein before and after modeling, and blood glucose was measured with a blood glucose meter. After fasting for 8 hours, the blood glucose concentration of the mice was tested. The results are shown in Tables 17 to 30 and Figure 4 The results showed that the blood sugar level of mice in the sea buckthorn group decreased significantly.

[0191] Table 17 Blood glucose of mice in low concentration administration group (C1-C8 groups) before administration

[0192]

[0193] Table 18 Blood glucose of mice in the low concentration administration group (C1-C8 groups) after administration for 1 week

[0194]

[0195] Table 19 Blood glucose of mice in the low concentration administration group (C1-C8 groups) after 2 weeks of administration

[0196]

[0197] Table 20 Blood glucose of mice in the low concentration administration group (C1-C8 groups) after 3 weeks of administration

[0198]

[0199]

[0200] Table 21 Blood glucose of mice in the low concentration administration group (C1-C8 groups) after 4 weeks of administration

[0201]

[0202] Table 22 Blood glucose of mice in the low concentration administration group (C1-C8 groups) after 5 weeks of administration

[0203]

[0204] Table 23 Blood glucose of mice in the low concentration administration group (C1-C8 groups) after 6 weeks of administration

[0205]

[0206]

[0207] Table 24 Blood glucose of mice in the high concentration administration group (D2-D7 groups) before administration

[0208]

[0209] Table 25 Blood glucose of mice in the high concentration administration group (D2-D7 groups) after administration for 1 week

[0210]

[0211]

[0212] Table 26 Blood glucose of mice in the high concentration administration group (D2-D7 groups) after 2 weeks of administration

[0213]

[0214] Table 27 Blood glucose of mice in the high concentration administration group (D2-D7 group) after 3 weeks of administration

[0215]

[0216]

[0217] Table 28 Blood glucose of mice in the high concentration administration group (D2-D7 groups) after 4 weeks of administration

[0218]

[0219] Table 29 Blood glucose of mice in the high concentration administration group (D2-D7 groups) after 5 weeks of administration

[0220]

[0221] Table 30 Blood glucose of mice in the high concentration administration group (D2-D7 group) after 6 weeks of administration

[0222] .

[0224] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A small molecule peptide mixture, characterized in that: The invention comprises the following raw materials for preparation: plant raw materials and plant lactic acid bacteria; the plant raw materials comprise the following raw materials in parts by weight: 5-10 parts of roxburghii, 8-12 parts of tuckahoe, 8-12 parts of wolfberry, 10-15 parts of walnut and 10-15 parts of acanthopanax senticosus; the plant lactic acid bacteria comprise Lactobacillus plantarum, Lactobacillus casei and Lactobacillus brevis.

2. The small molecule peptide mixture according to claim 1, characterized in that: The plant raw materials include the following raw materials in parts by mass: 5 parts of roxburghii, 8 parts of poria, 8 parts of wolfberry, 10 parts of walnuts and 10 parts of acanthopanax senticosus; or, the plant raw materials include the following raw materials in parts by mass: 10 parts of roxburghii, 12 parts of poria, 12 parts of wolfberry, 15 parts of walnuts and 15 parts of acanthopanax senticosus.

3. The small molecule peptide mixture according to claim 1, characterized in that: The plant lactobacillus includes plant lactobacillus ATCC8014; the casei lactobacillus includes casei lactobacillus BCC36987; and the brevis lactobacillus includes brevis lactobacillus D14349.

4. The method for preparing the small molecule peptide mixture according to any one of claims 1 to 3, characterized in that: The following steps are involved: mixing the plant material and water to obtain a mixed solution; The pH value of the mixed solution is adjusted to 5.8-6.2 and then sterilized to obtain a fermentation raw material; Inoculating plant lactic acid bacteria into the fermentation raw material, performing fermentation, and obtaining a fermentation product; The fermentation product is mixed with cellulase and subjected to a first enzymolysis to obtain a first enzymolysis product; The first enzymatic hydrolysis product is mixed with pectinase, and subjected to a second enzymatic hydrolysis to obtain a second enzymatic hydrolysis product; The second enzymatic hydrolysis product is mixed with papain, and subjected to a third enzymatic hydrolysis to obtain a third enzymatic hydrolysis product; The third enzymatic hydrolysis product is subjected to ultrafiltration, and the retentate is collected to obtain a small molecule peptide mixture; the ultrafiltration molecular weight cutoff is 2000Da.

5. The preparation method according to claim 4, characterized in that: The effective viable count of the plant lactic acid bacteria is ≥ 1×10 11 CFU / g; the inoculation amount of the plant lactic acid bacteria is 0.5-1g / L.

6. The preparation method according to claim 4, characterized in that: The temperature of the first enzymolysis is 55°C; the pH of the first enzymolysis is 6; the time of the first enzymolysis is 3 hours; the temperature of the second enzymolysis is 60°C; the pH of the second enzymolysis is 6.5; the time of the second enzymolysis is 3 hours; the temperature of the third enzymolysis is 60°C; the pH of the third enzymolysis is 6.5; the time of the third enzymolysis is 1.5 hours.

7. A lyophilized powder of a small molecule peptide mixture, characterized in that: It comprises a small molecule peptide mixture and a sweetener; the small molecule peptide mixture is the small molecule peptide mixture described in any one of claims 1 to 3 or the small molecule peptide mixture prepared by the preparation method described in any one of claims 4 to 6.

8. The method for preparing the freeze-dried powder according to claim 7, characterized in that: The following steps are involved: Mixing the small molecule peptide mixture, water and a sweetener to obtain a mixed solution; Sterilizing the mixed solution to obtain a sterilized solution; The sterilized liquid is sequentially frozen and vacuum-dehydrated to obtain a lyophilized powder of a small molecule peptide mixture.

9. Use of the small molecule peptide mixture according to any one of claims 1 to 3, the small molecule peptide mixture prepared by the preparation method according to any one of claims 4 to 6, the lyophilized powder according to claim 7, or the lyophilized powder prepared by the preparation method according to claim 8 in lowering blood sugar, assisting in lowering blood sugar or regulating body weight.

10. Use of the small molecule peptide mixture according to any one of claims 1 to 3, the small molecule peptide mixture prepared by the preparation method according to any one of claims 4 to 6, the lyophilized powder according to claim 7, or the lyophilized powder prepared by the preparation method according to claim 8 in improving memory or assisting in improving memory.

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