Earthworm polypeptide oral liquid and preparation method thereof

Abstract

The invention belongs to the technical field of natural composition preparation, and particularly relates to earthworm polypeptide oral liquid and a preparation method thereof. According to the invention, 15-20 parts of earthworm peptide powder and 5-8 parts of conch peptide powder are specifically combined, and the earthworm peptide powder in the composition is beneficial to promoting the utilization of sugar by muscles, so that the glycemic index is reduced; the conch peptide powder can regulate starvation hormone, reduce the proportion of pathogenic bacteria in intestinal tracts and improve the abundance of beneficial bacteria. Therefore, when the polypeptide composition is used for preparing the earthworm polypeptide oral liquid for resisting sugar and inhibiting harmful intestinal flora, the anti-sugar effect can be achieved, meanwhile, the increase of the number of enterobacter and / or enterococcus in the intestinal tract is effectively inhibited, and the occurrence of intestinal inflammation is reduced. Therefore, according to the technical scheme, the application range of the earthworms is enriched, and the application value of the market is widened.

Description

Technical Field

[0001] This invention belongs to the field of natural composition preparation technology, specifically relating to an earthworm polypeptide oral liquid and its preparation method. Background Technology

[0002] Earthworm (Pheretima) is a traditional Chinese medicine with important medicinal value. It is mainly derived from the dried whole body of worms of the family Phyllostachys, such as Phyllostachys spp., Phyllostachys wieniensis, Phyllostachys pubescens, and Phyllostachys pubescens. It has the effects of clearing heat and calming the nerves, promoting blood circulation and relieving asthma, promoting diuresis, and anticoagulation and thrombolysis.

[0003] Therefore, earthworms are often used in the preparation of anticoagulant and thrombolytic products. For example, Chinese patent CN 104004806B discloses the following: Earthworm polypeptide with anticoagulant and thrombolytic properties, its enzymatic preparation method, and its application. Specifically, the following steps are disclosed: Earthworm dried body is washed, dried, and pulverized into powder; water is added, stirred, and centrifuged to obtain protein precipitate; the protein is dissolved in water, and alkaline protease is added at a ratio of enzyme volume to protein weight of 0.1-3 mL:100 g, and stirred; enzyme activity is inactivated by adding trypsin at a ratio of enzyme mass to protein weight of 0.5-5 g:100 g, at a temperature of 25-60℃, and stirred; the enzymatic hydrolysate is passed through an anion exchange column, and a portion of the eluent is collected; then it is passed through a macroporous adsorption resin for separation and purification, collected, and dried to obtain earthworm anticoagulant peptide powder. This invention provides a new source of highly active anticoagulant substances, with advantages such as high product activity, convenient and feasible preparation process, and environmental friendliness. It can be used as a raw material for foods that prevent thrombosis, dissolve thrombi, and reduce blood viscosity, and can be supplemented with lactose, mannitol, etc., to make powders, tablets, capsules, or oral liquids.

[0004] However, with increasing health awareness, combating glycation and inhibiting harmful gut bacteria have become research hotspots. Currently, however, there are no reports of using earthworm peptides to prepare products for blood sugar control or inhibiting harmful gut bacteria. Therefore, if products with blood sugar control and harmful gut bacteria inhibition properties could be prepared from earthworms, it would unlock new uses for earthworms and broaden their market application value. Summary of the Invention

[0005] The purpose of this invention is to provide an earthworm polypeptide oral liquid and its preparation method. The oral liquid prepared in this way has the characteristics of controlling blood sugar and inhibiting harmful intestinal bacteria.

[0006] The present invention provides a polypeptide composition containing earthworm, wherein the composition comprises, by weight parts, the following raw materials: 15-20 parts earthworm peptide powder and 5-8 parts conch peptide powder.

[0007] Preferably, the preparation methods of the earthworm peptide powder or sea snail peptide powder respectively include: Perilla was mixed with water and then subjected to ultrasonic extraction to obtain perilla extract. The sample to be extracted, perilla extract and compound enzyme are mixed and enzymatically hydrolyzed, and the hydrolysate is concentrated to 1 / 8-1 / 9 of the original volume. The resulting extract is peptide powder. The samples to be extracted include: earthworms or sea snails; The complex enzyme includes trypsin and β-glucuronidase.

[0008] Preferably, the mass-to-volume ratio of perilla to water is 1.8-2.5 g: 10 mL; The parameters for ultrasonic extraction include: temperature of 80-85℃, time of 30-35min, and power of 1000-1200w.

[0009] Preferably, the mass ratio of trypsin to β-glucuronidase is 1:0.3-0.5.

[0010] Preferably, the mass-to-volume ratio of the sample to be extracted, the perilla extract, and the compound enzyme is 2-3g:100mL:0.5-0.8g; The parameters for the enzymatic hydrolysis include: temperature of 38-42℃ and time of 10-12h.

[0011] This invention provides the application of the polypeptide composition described above in the preparation of blood sugar control products and / or products that inhibit harmful intestinal flora.

[0012] Preferably, the harmful intestinal flora includes Enterobacteriaceae and / or Enterococci.

[0013] Preferably, the product includes: pharmaceuticals and / or health foods.

[0014] This invention provides an earthworm polypeptide oral liquid, which, by mass, comprises the following raw materials: 10-15 parts of the polypeptide composition described in the above technical solution, 1-3 parts of sodium benzoate, 2-5 parts of malic acid, and 100-120 parts of water.

[0015] This invention provides a method for preparing the earthworm polypeptide oral liquid described in the above technical solution, comprising the following steps: The polypeptide composition described in the above technical solution is mixed evenly with water to obtain a mixture; Sodium benzoate and malic acid were added to the mixture to obtain earthworm polypeptide oral liquid.

[0016] Beneficial effects: This invention provides a polypeptide composition containing earthworm, characterized in that, by weight parts, the composition comprises the following raw materials: 15-20 parts earthworm peptide powder and 5-8 parts conch peptide powder. The earthworm peptide powder provided by this invention promotes the utilization of glucose by muscles, thereby reducing the glycemic index; the conch peptide powder can regulate hunger hormones, reduce the proportion of pathogenic bacteria in the intestine, and increase the abundance of beneficial bacteria. Therefore, by specifically combining the two, this composition can be used in the preparation of products that are anti-glycation and inhibit harmful intestinal flora.

[0017] This invention also provides an earthworm polypeptide oral liquid, which, by weight, comprises the following raw materials: 10-15 parts of the polypeptide composition described in the above technical solution, 1-3 parts of sodium benzoate, 2-5 parts of malic acid, and 100-120 parts of water. This earthworm polypeptide oral liquid has a simple preparation method, high product activity, good taste, and is anti-glycemic, effectively inhibiting intestinal bacteria (Enterobacteria). Enterobacteriaceae ) and / or Enterococci ( Enterococcus The increase in quantity. Detailed Implementation

[0018] In this invention, unless otherwise specified, the raw materials, methods and equipment used are all conventional selections.

[0019] To further illustrate the present invention, the solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0020] Example 1 The steps for preparing a polypeptide composition containing earthworms are as follows: (1) Preparation of Perilla ( Perilla frutescens (L.) Britt. Extract Wash the perilla leaves thoroughly, dry them, and then crush them to obtain a powder (particle size ≤ 0.1 cm). The pulverized material was mixed with water at a ratio of 2.5g:10mL and ultrasonically extracted at 80℃ and 1150w for 30min to obtain perilla extract. (2) Preparation of earthworm peptide powder With ginseng ring-haired earthworm ( Pheretima aspergillus The dried whole is the raw material; The above raw materials are cleaned and crushed to obtain crushed earthworms (the particle size of the crushed earthworms is ≤0.1cm). Trypsin and β-glucuronidase were mixed evenly at a ratio of 1:0.5 to obtain a composite enzyme; Take 25g of crushed earthworm, 1000mL of perilla extract and 5g of compound enzyme and mix them together. Enzymatically hydrolyze at 40℃ for 12h. During the enzymatic hydrolysis, stir every 3 hours to obtain the enzymatic hydrolysate. After passing the enzymatic hydrolysate through a 200-mesh sieve, the enzymatic hydrolysate is obtained. The enzymatic hydrolysate is concentrated to 1 / 8 of the original volume to obtain earthworm peptide powder. (3) Preparation of conch peptide powder Fresh whelks ( Neptune cuming Cross The meat is used as the raw material; The above raw materials are cleaned and crushed to obtain crushed snails (the particle size of the crushed earthworms is ≤0.1cm). Trypsin and β-glucuronidase were mixed evenly at a ratio of 1:0.3 to obtain a composite enzyme; Take 25g of crushed whelk, 1000mL of perilla extract and 5g of compound enzyme and mix them together. Enzymatically hydrolyze at 40℃ for 12h. During the enzymatic hydrolysis, stir every 3 hours to obtain the enzymatic hydrolysate. After passing the enzymatic hydrolysate through a 200-mesh sieve, the enzymatic hydrolysate is obtained. The enzymatic hydrolysate is concentrated to 1 / 8 of the original volume to obtain whelk peptide powder. (4) Polypeptide compositions containing earthworm By weight, 15 parts of earthworm peptide powder and 5 parts of conch peptide powder were mixed evenly to obtain a composition.

[0021] Example 2 The steps for preparing a polypeptide composition containing earthworms are as follows: (1) The method is the same as step (1) in Example 1; (2) The method is the same as step (2) in Example 1; (3) The method is the same as step (3) in Example 1; (4) Polypeptide compositions containing earthworm By weight, 16 parts of earthworm peptide powder and 8 parts of conch peptide powder were mixed evenly to obtain a composition.

[0022] Comparative Example 1 The steps for preparing a polypeptide composition containing earthworms are as follows: (1) Step (1) is omitted; (2) The method is the same as step (2) in Example 1, but the perilla extract in Example 1 is replaced with water to prepare earthworm peptide powder; (3) The same method as step (3) in Example 1, but the perilla extract in Example 1 is replaced with water to prepare conch peptide powder; (4) is the same as step (4) in Example 1.

[0023] Comparative Example 2 The steps for preparing a polypeptide composition containing earthworms are as follows: (1) The method is the same as step (1) in Example 1; (2) Preparation of earthworm peptide powder With ginseng ring-haired earthworm ( Pheretima aspergillus The dried whole is the raw material; The above raw materials are cleaned and crushed to obtain crushed earthworms (the particle size of the crushed earthworms is ≤0.1cm). Take 25g of crushed earthworm, 1000mL of perilla extract and 5g of trypsin and mix them together. Incubate at 40℃ for 12h. Stir every 3 hours during the enzymatic hydrolysis to obtain the hydrolysate. Pass the hydrolysate through a 200-mesh sieve to obtain the hydrolysate. Concentrate the hydrolysate to 1 / 8 of the original volume to obtain earthworm peptide powder. (3) Preparation of conch peptide powder Fresh whelks ( Neptune cuming Cross The meat is used as the raw material; The above raw materials are cleaned and crushed to obtain crushed snails (the particle size of the crushed earthworms is ≤0.1cm). Take 25g of crushed whelk, 1000mL of perilla extract and 5g of trypsin and mix them together. Incubate at 40℃ for 12h. Stir every 3 hours during the enzymatic hydrolysis to obtain the hydrolysate. After passing the hydrolysate through a 200-mesh sieve, obtain the hydrolysate solution. Concentrate the hydrolysate solution to 1 / 8 of the original volume to obtain whelk peptide powder. (4) is the same as step (4) in Example 1.

[0024] Comparative Example 3 The steps for preparing a polypeptide composition containing earthworms are as follows: (1) Preparation of ginseng ( Panax ginseng CA Mey Extract Clean the ginseng, dry it, and then grind it into powder (particle size ≤ 0.1 cm). The pulverized material was mixed with water at a ratio of 2.5g:10mL and ultrasonically extracted at 80℃ and 1150w for 30min to obtain ginseng extract. (2) The method is the same as step (2) in Example 1, but the perilla extract in Example 1 is replaced with ginseng extract to prepare earthworm peptide powder; (3) The same method as step (3) in Example 1, but the perilla extract in Example 1 is replaced with ginseng extract to prepare conch peptide powder; (4) is the same as step (4) in Example 1.

[0025] Comparative Example 4 The steps for preparing earthworm peptide powder are as follows: (1) The method is the same as step (1) in Example 1; (2) Preparation of earthworm peptide powder With ginseng ring-haired earthworm ( Pheretima aspergillus The dried whole is the raw material; The above raw materials are cleaned and crushed to obtain crushed earthworms (the particle size of the crushed earthworms is ≤0.1cm). Trypsin and β-glucuronidase were mixed evenly at a ratio of 1:0.5 to obtain a composite enzyme; Take 25g of crushed earthworm, 1000mL of perilla extract and 5g of compound enzyme and mix them together. Enzymatically hydrolyze at 40℃ for 12h. During the enzymatic hydrolysis, stir every 3 hours to obtain the enzymatic hydrolysate. After passing the enzymatic hydrolysate through a 200-mesh sieve, the enzymatic hydrolysate is obtained. The enzymatic hydrolysate is concentrated to 1 / 8 of the original volume to obtain earthworm peptide powder. Steps (3) and (4) are omitted.

[0026] Comparative Example 5 The steps for preparing a polypeptide composition containing earthworms are as follows: (1) Preparation of perilla extract; Wash the perilla leaves thoroughly, dry them, and then crush them to obtain a powder (particle size ≤ 0.1 cm). The pulverized material was mixed with water at a ratio of 2.5g:10mL and ultrasonically extracted at 80℃ and 1150w for 30min. The extract was then subjected to reduced pressure to obtain perilla extract. (2) Preparation of earthworm peptide powder With ginseng ring-haired earthworm ( Pheretima aspergillus The dried whole is the raw material; The above raw materials are cleaned and crushed to obtain crushed earthworms (the particle size of the crushed earthworms is ≤0.1cm). Trypsin and β-glucuronidase were mixed evenly at a ratio of 1:0.5 to obtain a composite enzyme; Take 25g of crushed earthworm, 1000mL of water and 5g of compound enzyme and mix them. Incubate at 40℃ for 12h. Stir every 3 hours during the incubation process to obtain the enzymatic hydrolysate. After passing the enzymatic hydrolysate through a 200-mesh sieve, the enzymatic hydrolysate is obtained. Concentrate the enzymatic hydrolysate to 1 / 8 of the original volume to obtain earthworm peptide powder. (3) Polypeptide composition containing earthworm By weight, 15 parts of earthworm peptide powder and 5 parts of perilla extract were mixed evenly to obtain the composition.

[0027] Application Example 1: Anti-glucose test Several healthy male rats with similar weights (190-200g) were selected and randomly divided into an experimental group, a negative control group, and a positive control group, with 10 rats in each group. The experimental period was 4 weeks. All groups were raised in the same environment, with the only difference being the type of feed.

[0028] Negative control group: fed with standard ordinary feed; Positive control group: fed a high-sugar diet (the high-sugar diet consisted of ordinary feed, lard, sucrose, cholesterol and sodium cholate in a mass ratio of 67:10:20:2.5:0.5). Experimental Group 1: Feeded with feed 1 (feed 1 consisted of a high-sugar feed and the polypeptide composition prepared in Example 1 at a mass ratio of 97:3); Experimental Group 2: Feed 2 (feed 2 consisted of a high-sugar feed and the polypeptide composition prepared in Example 2 at a mass ratio of 97:3); Experimental Group 3: Feed 3 (Feed 6 consisted of a high-sugar feed and the polypeptide composition prepared in Comparative Example 1 at a mass ratio of 97:3); Experimental Group 4: Feed 3 (Feed 6 consisted of a high-sugar feed and the polypeptide composition prepared in Comparative Example 2 at a mass ratio of 97:3); Experimental Group 5: Feeded with feed 3 (feed 6 consisted of a high-sugar feed and the polypeptide composition prepared in Comparative Example 3 at a mass ratio of 97:3); Experimental Group 6: fed with feed 3 (feed 6 consisted of a high-sugar feed and the polypeptide composition prepared in Comparative Example 4 at a mass ratio of 97:3); Experimental Group 7: Feeded with feed 3 (feed 6 consisted of a high-sugar feed and the polypeptide composition prepared in Comparative Example 5 at a mass ratio of 97:3); After the experiment, the mean fasting blood glucose (i.e., fasting blood glucose before the last feeding) and the mean blood glucose 2 hours after the last feeding were measured in each treatment. The results are shown in Table 1.

[0029] Table 1. Blood glucose levels in rats under different treatments

[0030] Based on the data in Table 1, it can be seen that the polypeptide compositions prepared in Example 1 (Experiment 1 Group) and Example 2 (Experiment 2 Group) have good anti-glycation effects; while the polypeptide compositions prepared in Comparative Example 1 (Experiment 3 Group) and Comparative Example 3 (Experiment 5 Group) have better anti-glycation effects. Therefore, the polypeptide compositions with better anti-glycation effects will be used for subsequent experimental verification.

[0031] Application Example 2: Test to Inhibit Harmful Bacteria The compositions prepared in Examples 1, 2, 1, and 3 were used to prepare earthworm polypeptide oral liquid for experimental verification. The process is as follows: Group 1: Take 1g of the polypeptide composition prepared in Example 1 and mix it with 10g of water to obtain a mixture; add 0.1g of sodium benzoate and 0.2g of malic acid to the mixture and mix well to obtain earthworm polypeptide oral liquid; Group 2: Take 1g of the polypeptide composition prepared in Example 2 and mix it with 10g of water to obtain a mixture; add 0.1g of sodium benzoate and 0.2g of malic acid to the mixture and mix well to obtain earthworm polypeptide oral liquid; Group 3: Take 1g of the polypeptide composition prepared in Comparative Example 1 and mix it with 10g of water to obtain a mixture; add 0.1g of sodium benzoate and 0.2g of malic acid to the mixture and mix well to obtain earthworm polypeptide oral liquid; Group 4: Take 1g of the polypeptide composition prepared in Comparative Example 3 and mix it with 10g of water to obtain a mixture; add 0.1g of sodium benzoate and 0.2g of malic acid to the mixture and mix well to obtain earthworm polypeptide oral liquid; Several healthy male mice weighing 19-21g were selected and randomly divided into an experimental group and a negative control group, with 10 mice in each group. The experimental period was 14 days. All groups were fed conventional feed and kept in the same environment.

[0032] Group 1: Each mouse was administered 0.1 mL of the first group's earthworm polypeptide oral solution by gavage 2 hours after its first meal each day; Experimental Group 2: Each mouse was administered 0.1 mL of the second group's earthworm polypeptide oral solution by gavage 2 hours after its first meal each day; Experimental Group 3: Each mouse was administered 0.1 mL of the third group's earthworm polypeptide oral solution by gavage 2 hours after its first meal each day; Group 4 of the experiment: Each mouse was given 0.1 mL of earthworm polypeptide oral solution (Group 4) by gavage 2 hours after its first meal each day; Negative control group: Each mouse was given 0.1 mL of water by gavage 2 hours after its first meal each day; Before the first gavage, feces from mice in each treatment group were aseptically collected and prepared into bacterial suspensions for preservation. After the 14-day experiment, feces from mice in each treatment group were aseptically collected and prepared into bacterial suspensions. The number of Enterobacteriaceae and Enterococci in each treatment group was determined by the serial dilution method. The results are shown in Table 2.

[0033] Table 2. Gut microbiota counts in mice under different treatments

[0034] Enterobacteriaceae and Enterococci can cause intestinal inflammation and dysbiosis, and are considered harmful bacteria in the gut. As shown in Table 2, the earthworm polypeptide oral solution prepared in Examples 1 (Experiment 1) and 2 (Experiment 2) effectively inhibited the proliferation of Enterobacteriaceae and Enterococci, with an inhibition rate of 44.4%-45.2%; while the earthworm polypeptide oral solution prepared in the comparative example was not effective in inhibiting harmful bacteria. Therefore, the earthworm polypeptide oral solution prepared in this invention has the effect of inhibiting harmful intestinal bacteria and optimizing the intestinal flora.

[0035] In summary, this invention provides a polypeptide composition containing earthworm. By specifically combining 15-20 parts of earthworm peptide powder and 5-8 parts of conch peptide powder, the earthworm peptide powder in the composition promotes muscle glucose utilization, thereby lowering the glycemic index; the conch peptide powder can regulate ghrelin, reduce the proportion of pathogenic bacteria in the intestine, and increase the abundance of beneficial bacteria. Therefore, when this polypeptide composition is used in the preparation of an anti-glycation, anti-intestinal harmful bacteria oral liquid containing earthworm peptides, it can release an anti-glycation effect while effectively inhibiting the increase in the number of Enterobacteriaceae and / or Enterococci in the intestine, reducing the occurrence of intestinal inflammation.

[0036] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A polypeptide composition containing earthworm, characterized in that, The composition comprises, by weight parts, the following raw materials: 15-20 parts earthworm peptide powder and 5-8 parts conch peptide powder.

2. The polypeptide composition according to claim 1, characterized in that, The preparation methods of the earthworm peptide powder or sea snail peptide powder respectively include: Perilla was mixed with water and then subjected to ultrasonic extraction to obtain perilla extract. The sample to be extracted, perilla extract and compound enzyme are mixed and enzymatically hydrolyzed, and the hydrolysate is concentrated to 1 / 8-1 / 9 of the original volume. The resulting extract is peptide powder. The samples to be extracted include: earthworms or sea snails; The complex enzyme includes trypsin and β-glucuronidase.

3. The polypeptide composition according to claim 2, characterized in that, The mass-to-volume ratio of perilla to water is 1.8-2.5 g: 10 mL; The parameters for ultrasonic extraction include: temperature of 80-85℃, time of 30-35min, and power of 1000-1200w.

4. The polypeptide composition according to claim 2, characterized in that, The mass ratio of trypsin to β-glucuronidase is 1:0.3-0.

5.

5. The polypeptide composition according to claim 2 or 4, characterized in that, The mass-to-volume ratio of the sample to be extracted, perilla extract, and compound enzyme is 2-3g:100mL:0.5-0.8g; The parameters for the enzymatic hydrolysis include: temperature of 38-42℃ and time of 10-12h.

6. The use of the polypeptide composition of claim 1 in the preparation of blood sugar control products and / or products that inhibit harmful intestinal flora.

7. The application according to claim 6, characterized in that, The harmful intestinal flora includes Enterobacteriaceae and / or Enterococci.

8. The application according to claim 6, characterized in that, The products include: pharmaceuticals and / or health foods.

9. An earthworm polypeptide oral liquid, characterized in that, The earthworm polypeptide oral liquid comprises the following raw materials in parts by weight: 10-15 parts of the polypeptide composition according to any one of claims 1-5, 1-3 parts of sodium benzoate, 2-5 parts of malic acid, and 100-120 parts of water.

10. The method for preparing the earthworm polypeptide oral liquid according to claim 9, characterized in that, Includes the following steps: The polypeptide composition according to any one of claims 1-5 is mixed with water to obtain a mixture; Sodium benzoate and malic acid were added to the mixture to obtain earthworm polypeptide oral liquid.

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