Preparation method of maggot active peptide
Through grinding and enzyme extract, the dehydrated scum powder is solved, the problem of complexity and small amount of obtaining the extraction process of pentagram activated peptide peptide peptide in the existing technologies has been solved.
Patent Information
- Application Number
- CN202310917993.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-07-25
- Publication Date
- 2025-01-28
AI Technical Summary
In the prior art, the extraction process of grain active peptides is complex, the amount obtained is small, and it is difficult to produce industrially, which restricts the application and promotion of grain active peptides.
By grinding the dried pentaidine scum powder into fines, add water to dissolve after the sieve, adjust the pH value, add enzyme solution, extract active peptide under the water bath, and finally obtain the pentae active peptide product through filtering and frozen drying.
The gain rate of grapepel activity peptides has been increased, the preparation technology is simplified, the cost of production is reduced, and it provides the possibility of the industrial production and application promotion of the grapemine activated peptide.
Smart Images
Figure BDA0004357638170000041 
Figure BDA0004357638170000061
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of medicines, and in particular to a method for preparing a five-grain insect active peptide. Background Art
[0002] The source of the grain worm is the larvae or pupae of the green fly or the big-headed golden fly of the Calliphoridae family. They are both medicinal grain worms and have the same medicinal properties. According to the "Source of Materia Medica" and "Compendium of Materia Medica", after being processed and ground into powder, the grain worm can be applied externally to treat surgical infectious diseases such as bedsores and lip boils. Traditional Chinese medicine believes that the medicinal properties of the grain worm are cold, and it has the effects of clearing away heat and stagnation, detoxifying and anti-inflammatory, removing dead tissue and promoting tissue regeneration. It can also treat infectious surgical diseases such as bedsores. The current use of medicinal grain worms is mainly for clinical debridement treatment in living bodies, but living grain worms have problems such as low patient acceptance and difficulty in breeding and preservation, making them difficult to promote and use.
[0003] In recent years, researchers have conducted a lot of research on the active substances of Chinese medicine five grain insects. Studies have found that five grain insect fatty acids, five grain insect secretions, and five grain insect dry powder have a wide range of pharmacological activities, such as antibacterial / antibacterial, wound healing, anti-tumor, anti-apoptosis, and antioxidant. The peptide components secreted by five grain insects are endogenous components with good activity. Some researchers have found that active peptides have the characteristics of wide biological activity, strong specificity, good stability, and few toxic and side effects. However, the extraction process of five grain insect active peptides in the prior art is complicated, and the amount of active peptides obtained is small, which is difficult to industrialize, restricting the application and promotion of five grain insect active peptides. Therefore, it is necessary to find a method for preparing a five grain insect active peptide to increase the yield of five grain insect active peptides. Summary of the invention
[0004] The present invention provides a method for preparing five cereal insect active peptides to solve the above problems.
[0005] In order to achieve the above object, the technical solution of the present invention is:
[0006] A method for preparing five cereal insect active peptides, comprising the following preparation steps:
[0007] S1: Take the dried and defatted grain insect residue powder and put it into a mortar and grind it thoroughly;
[0008] S2: Grind the dried defatted grain worm residue powder into fine powder and then sieve it to obtain defatted grain worm fine powder;
[0009] S3: Take the defatted fine powder of grain worms obtained in step S2 as a substrate, add water to dissolve it, stir to mix it evenly, and boil it in a water bath;
[0010] S4: Adjust the solution pH to 6-7, add enzyme solution, and extract active peptides under water bath conditions;
[0011] S5: Filter through a membrane, and freeze-dry the filtrate to obtain the five cereal insect active peptide product.
[0012] Preferably, the dried defatted grain worm residue powder is ground and passed through a 40-70 mesh sieve.
[0013] Preferably, the mass volume ratio of the substrate concentration is 0.5-10%, and the boiling time in a water bath is 10-60 minutes.
[0014] Preferably, the method for adjusting the pH to 6-7 is to add 8-12% HCl or 8-12% phosphoric acid to the solution after water bath for adjustment.
[0015] Preferably, the enzyme activity of the enzyme solution is 3000-10000 U / g, the temperature of water bath extraction is 30-70° C., and the extraction time is 1-8 h.
[0016] Preferably, the source of the defatted grain insect residue is the defatted dried larvae of Muscidae or Calliphoridae. In practical applications, the dried larvae of other closely related insects can also be used.
[0017] Preferably, the enzyme used is one of papain, alkaline protease, trypsin and pepsin.
[0018] The beneficial effects of the present invention are:
[0019] The invention discloses a method for preparing the active peptide of five cereal insects, which improves the resource utilization rate of the traditional Chinese medicine five cereal insects, has a simple preparation method, reduces the production cost, and has a high yield of the active peptide of five cereal insects extracted by the preparation method, thus providing the possibility for the industrialized production and application promotion of the active peptide of five cereal insects. DETAILED DESCRIPTION
[0020] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0021] Embodiment 1:
[0022] Take an appropriate amount of dry defatted grain worm residue powder and put it into a mortar for grinding. After fully grinding into fine powder, filter it through a 50-mesh sieve to obtain defatted grain worm fine powder. Take 0.5g of defatted grain worm fine powder and dissolve it in 100ml of water to obtain a solution. Stir at room temperature for 4h and boil it in a water bath for 10min. After the solution returns to room temperature, adjust the pH to 6-7, add trypsin solution with an enzyme activity of 6000U / g, extract it under 50℃ water bath conditions for 5h, filter it through a 3KD filter membrane after cooling, and freeze-dry the filtrate to obtain the product; the active peptide yield is 32.8%.
[0023] Embodiment 2:
[0024] Take an appropriate amount of dry defatted grain worm residue powder and put it into a mortar for grinding. After it is fully ground into fine powder, filter it through a 50-mesh sieve to obtain defatted grain worm fine powder. Take 1.5g of defatted grain worm fine powder and dissolve it in 100ml of water to obtain a solution. Stir at room temperature for 4h and boil it in a water bath for 10min. After the solution returns to room temperature, adjust the pH to 6-7, add trypsin solution with an enzyme activity of 6000U / g, extract it under 50℃ water bath conditions for 6h, filter it through a 3KD filter membrane after cooling, and freeze-dry the filtrate to obtain the product; the active peptide yield is 20.3%.
[0025] Embodiment 3:
[0026] Take an appropriate amount of dry defatted grain worm residue powder and put it into a mortar for grinding. After fully grinding into fine powder, filter it through a 50-mesh sieve to obtain defatted grain worm fine powder. Take 1.0g of defatted grain worm fine powder and dissolve it in 100ml of water to obtain a solution. Stir at room temperature for 4h and boil it in a water bath for 10min. After the solution returns to room temperature, adjust the pH to 6-7, add trypsin solution with an enzyme activity of 6000U / g, extract it under 30℃ water bath conditions for 5h, filter it through a 3KD filter membrane after cooling, and freeze-dry the filtrate to obtain the product; the active peptide yield is 23.9%.
[0027] Embodiment 4:
[0028] Take an appropriate amount of dry defatted grain worm residue powder and put it into a mortar for grinding. After fully grinding into fine powder, filter it through a 50-mesh sieve to obtain defatted grain worm fine powder. Take 1.0g of defatted grain worm fine powder and dissolve it in 100ml of water to obtain a solution. Stir at room temperature for 4h and boil it in a water bath for 10min. After the solution returns to room temperature, adjust the pH to 6-7, add trypsin solution with an enzyme activity of 6000U / g, extract it under 70℃ water bath conditions for 6h, filter it through a 3KD filter membrane after cooling, and freeze-dry the filtrate to obtain the product; the active peptide yield is 8.9%.
[0029] Experimental Example 1: Study on the preparation process of five-grain insect active peptide
[0030] Experimental method: The three factors that have the most significant impact on the enzymatic extraction of grain worm polypeptides were selected as independent variables, namely: temperature (℃), time (h), and substrate concentration (%). The grain worm polypeptide extraction rate (Y) was taken as the response result. The Design-Expert 12 software was used to simulate the experiment. The node results were random. The final experiment had a total of 17 test points. The results are shown in Table 1.
[0031] Table 1 Response surface (Box-Behnken) experimental design results
[0032]
[0033] According to the experimental data in Table 1, the enzymatic hydrolysis simulation regression equation is obtained by mathematical analysis and statistical calculation by software as follows:
[0034] Y=36.86-2.37A-3.82B+0.285C+0.4091AB-0.025AC-0.005BC-14.46A 2 -8.21B 2 -4.19C 2
[0035] As shown in Table 2, the lack of fit of the quadratic multiple regression equation is not statistically significant (P = 0.4654> 0.05), and the regression model is significantly consistent (P < 0.0001). The statistical results strongly illustrate the credibility and accuracy of the experimental results, and the error factor is generated by random experiments, so the fitting equation has a higher credibility.
[0036] From Table 2, the correlation coefficient R of the mathematical statistical regression equation 2 =0.9966. The fit between the model and the experimental results is reliable, and the correction factor R 2 adj=0.9922.
[0037] It can be seen from Table 2 that in the multiple regression equation, A-temperature, B-time, A 2 , C 2 The effect on the response value (active peptide extraction rate) was significant (P<0.05). The other items were not significant. The data characteristics reflected by this result phenomenon are that the relationship between the factors in the reaction system is not positively correlated or negatively correlated and has a complex interactive nature.
[0038] The F value of each condition in Table 2 can also be used to directly compare the degree of effect of the condition on the extraction rate. The larger the F value, the more significant the influence of the factor. According to the F value, the influence of each factor on the extraction rate of grain insect polypeptide is ranked as follows: B-time>A-temperature>C-substrate concentration.
[0039] Table: 2 Response surface quadratic multivariate model variance analysis for grain insect polypeptide process optimization
[0040]
[0041] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.
Claims
1. A method for preparing a five-grain insect active peptide, characterized in that: The method comprises the following preparation steps: S1: Grind the dried grain insect defatted medicine residue powder; S2: sieving the ground dried five-grain insect defatted medicine residue powder to obtain five-grain insect defatted fine powder; S3: taking the defatted fine powder of grain worms obtained in step S2 as a substrate, adding water to dissolve it into a solution, and boiling it in a water bath; S4: Adjust the solution pH to 6-7, add enzyme solution, and extract active peptides in a water bath; S5: Filter, and freeze-dry the filtrate to obtain the five cereal insect active peptide product.
2. The method for preparing the active peptide from five cereal insects according to claim 1, characterized in that: In step S2, the dried defatted grain worm residue powder is ground and then passed through a 40-70 mesh sieve.
3. The method for preparing the active peptide from grain insects according to claim 1, characterized in that: In step S3, the mass volume ratio of the substrate concentration is 0.5-10%, and the boiling time in a water bath is 10-60 minutes.
4. The method for preparing the active peptide from five cereal insects according to claim 1, characterized in that: In step S4, the method for adjusting the pH to 6-7 is: adding 8-12% HCl or 8-12% phosphoric acid to the solution after the water bath for adjustment.
5. The method for preparing the active peptide from five cereal insects according to claim 1, characterized in that: In step S4, the enzyme activity of the enzyme solution is 3000-10000 U / g, the temperature of water bath extraction is 30-70° C., and the extraction time is 1-8 hours.
6. The method for preparing the active peptide from five cereal insects according to claim 1, characterized in that: The defatted grain insect residues are sourced from defatted dried larvae of the Muscidae family or Calliphoridae family.
7. The method for preparing the active peptide from five cereal insects according to claim 1, characterized in that: In step S4, the enzyme used is one of papain, alkaline protease, trypsin and pepsin.