CN113101305B - Fermented earthworm powder and preparation method thereof - Google Patents

Fermented earthworm powder and preparation method thereof Download PDF

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CN113101305B
CN113101305B CN202110269887.4A CN202110269887A CN113101305B CN 113101305 B CN113101305 B CN 113101305B CN 202110269887 A CN202110269887 A CN 202110269887A CN 113101305 B CN113101305 B CN 113101305B
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earthworm
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phosphate
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CN113101305A (en
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周希云
任利霞
任立安
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Beijing Aikesaide Bioengineering Co ltd
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Abstract

The application provides fermented earthworm powder and a preparation method thereof, and the fermented earthworm powder comprises the following steps: grinding earthworms for 100-140 minutes, gradually increasing the grinding temperature from 60-62 ℃ to 78-80 ℃, adding water during grinding, and obtaining earthworm grinding fluid after grinding; adding a mixing agent into the earthworm grinding liquid, fermenting at 78-82 ℃, wherein the mixing agent comprises 66-70% of staphylococcus carnosus and 30-34% of neutral protease by mass, and performing fermentation and enzymolysis to obtain earthworm fermentation liquid; adding phosphate into the earthworm fermentation liquor to adjust the pH value to 4.5-6.5, and keeping the earthworm fermentation liquor stable; drying the earthworm fermentation liquor added with phosphate to prepare powder, thus obtaining fermented earthworm powder. Through proper grinding temperature and grinding time, the protein is stably decomposed, the release of active ingredients and beneficial ingredients of the earthworms is promoted, and the earthworms also have broad-spectrum antibacterial performance.

Description

Fermented earthworm powder and preparation method thereof
Technical Field
The application relates to the field of polypeptide preparation, in particular to fermented earthworm powder and a preparation method thereof.
Background
Earthworms, also known as earthworms, are regarded more and more as being important because of their high edible and medicinal value. The earthworm powder prepared from the earthworms is used as a novel animal protein raw material, and has the advantages of easily obtained materials and low cost. The traditional earthworm powder processing method comprises a negative dry method (air dry method), a sunlight exposure method, a high-temperature drying method and an enzymolysis method, but the prepared earthworm powder has low retention of active ingredients and beneficial ingredients.
Disclosure of Invention
The application provides a preparation method of fermented earthworm powder, which aims to solve the problem that the earthworm powder has low retention of active ingredients and beneficial ingredients.
In a first aspect, an embodiment of the present application provides a method for preparing fermented earthworm powder, including the following steps:
grinding earthworms for 100-140 minutes, gradually increasing the grinding temperature from 60-62 ℃ to 78-80 ℃, adding water during grinding, and obtaining earthworm grinding fluid after grinding;
adding a mixing agent into the earthworm grinding liquid, fermenting at 78-82 ℃, wherein the mixing agent comprises 66-70% of staphylococcus carnosus and 30-34% of neutral protease by mass, and performing fermentation and enzymolysis to obtain earthworm fermentation liquid;
adding phosphate into the earthworm fermentation liquor to adjust the pH value to 4.5-6.5, and keeping the earthworm fermentation liquor stable;
drying the earthworm fermentation liquor added with phosphate to prepare powder, thus obtaining fermented earthworm powder.
According to one embodiment of the application, the grinding temperature is gradually increased from 60 ℃ to 80 ℃ within 55-65 minutes, the grinding comprises a first grinding stage in the temperature increasing process and a second grinding stage after the temperature increasing process, and the total grinding time is 120 minutes.
According to one embodiment of the application, the mixture comprises 68% by mass of staphylococcus carnosus and 32% by mass of protease.
According to one embodiment of the application, the addition amount of the mixture is 15-25% of the mass of the earthworm grinding fluid.
According to one embodiment of the application, the phosphate is a mixture of dipotassium hydrogen phosphate and potassium dihydrogen phosphate or a mixture of disodium hydrogen phosphate and sodium dihydrogen phosphate.
According to one embodiment of the application, the phosphate is a mixture of dipotassium hydrogen phosphate and potassium dihydrogen phosphate, the dosage of the dipotassium hydrogen phosphate is 3-6% of the mass of the earthworm fermentation liquor, and the dosage of the potassium dihydrogen phosphate is 1-3% of the mass of the earthworm fermentation liquor.
According to one embodiment of the present application, the phosphate is a mixture of disodium hydrogen phosphate and sodium dihydrogen phosphate, the amount of the disodium hydrogen phosphate is 3-6% of the mass of the earthworm fermentation broth, and the amount of the sodium dihydrogen phosphate is 1-3% of the mass of the earthworm fermentation broth.
According to one embodiment of the application, water is added in two times during grinding, and the total water addition amount is 2-2.6 times of the mass of the earthworms.
According to one embodiment of the application, the earthworm fermentation liquor added with phosphate is mixed with a carrier, and the mixture is sprayed and dried to form powder, wherein the adding amount of the carrier is more than 10% of the mass of the earthworm fermentation liquor added with phosphate.
In a second aspect, the present application provides a fermented earthworm powder prepared by the above preparation method.
According to the preparation method of the fermented earthworm powder, the protein is stably decomposed and the release of active ingredients and beneficial ingredients of the earthworms is promoted through proper grinding temperature and grinding time. During the fermentation process, the staphylococcus carnosus can decompose and convert earthworm self protein into peptide bond protein, and the smell of the fermented finished product is kept; the protease accelerates the hydrolysis of earthworm protein and promotes the effect of staphylococcus carnosus. And the active ingredients and beneficial ingredients of the earthworms are fully released, so that the protein of the fermented earthworm powder prepared by the preparation method of the fermented earthworm powder is fully decomposed into amino acid, the protein content of the fermented earthworm powder is 2-5%, the amino acid content is improved, and the earthworm powder also has the bacteriostatic performance of broad-spectrum antibiosis.
Drawings
Features, advantages and technical effects of exemplary embodiments of the present application will be described below with reference to the accompanying drawings.
FIG. 1 is a graph showing the results of bacteriostatic tests on Staphylococcus aureus using solutions of different concentrations disclosed in the examples of the present application;
FIG. 2 is a graph showing the results of bacteriostatic tests on Bacillus subtilis with solutions of different concentrations disclosed in the examples of the present application;
FIG. 3 is a graph showing the results of the bacteriostatic test of the solutions with different concentrations on Pseudomonas aeruginosa according to the embodiment of the present application;
FIG. 4 is a graph showing the results of the bacteriostatic test of different concentrations of the solutions against enterococcus faecalis disclosed in the examples of the present application;
FIG. 5 is a graph showing the results of the bacteriostatic test of the solutions with different concentrations on Escherichia coli disclosed in the examples of the present application.
Detailed Description
Embodiments of the present application will be described in further detail below with reference to the drawings and examples. The following detailed description of the embodiments and the accompanying drawings are provided to illustrate the principles of the application and are not intended to limit the scope of the application, i.e., the application is not limited to the described embodiments.
In the description of the present application, it is to be noted that, unless otherwise specified, "a plurality" means two or more; the terms "upper," "lower," "left," "right," "inner," "outer," and the like, indicate an orientation or positional relationship that is merely for convenience in describing the application and to simplify the description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be construed as limiting the application. Furthermore, the terms "first," "second," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance. "vertical" is not strictly vertical, but is within the tolerance of the error. "parallel" is not strictly parallel but within the tolerance of the error.
In the description of the present application, it is also to be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; may be directly connected or indirectly connected through an intermediate. The specific meaning of the above terms in the present application can be understood as appropriate by one of ordinary skill in the art.
The embodiment of the application provides a preparation method of fermented earthworm powder, which comprises the following steps:
grinding the earthworms for 100-140 minutes, gradually increasing the grinding temperature from 60-62 ℃ to 78-80 ℃, adding water during grinding, and obtaining the earthworm grinding fluid after grinding.
Adding a mixing agent into the earthworm grinding liquid, fermenting at 78-82 ℃, wherein the mixing agent comprises 66-70% of staphylococcus carnosus and 30-34% of neutral protease by mass, and performing fermentation and enzymolysis to obtain earthworm fermentation liquid.
And (3) adding phosphate into the earthworm fermentation liquor to adjust the pH value to 4.5-6.5, and keeping the stability of the earthworm fermentation liquor.
Drying the earthworm fermentation liquor added with phosphate to prepare powder, thus obtaining fermented earthworm powder.
In the grinding step, the earthworms may be selected from fresh earthworms, such as fresh earthworms cultured, harvested and processed according to a standard. The milling may be carried out in a ball mill to break down the earthworms. The earthworms can be ground at 60-62 ℃, water is added during grinding, and the temperature is gradually increased to 78-80 ℃ during grinding. When the temperature is increased to 78-80 ℃, the grinding is continued for a period of time. The total grinding time is 100-140 minutes.
The temperature of 60-62 ℃ is the temperature at which the earthworm protein can be decomposed, and the temperature of 78-80 ℃ is the temperature at which the decomposition speed of the earthworm protein is higher. And the temperature is gradually increased, so that the decomposition speed of the earthworm protein is gradually increased. And grinding for a period of time at 78-80 ℃ to ensure that the earthworm protein is fully extracted and stably decomposed and promote the release of active ingredients and beneficial ingredients of the earthworms.
Adding a mixing agent into the separated liquid, and fermenting at 78-82 ℃. The mixture contains Staphylococcus carnosus and protease, so that the decomposition and enzymolysis of earthworm protein can be promoted. During the fermentation process, the staphylococcus carnosus can decompose and convert earthworm self protein into peptide bond protein, and the smell of the fermented finished product is kept; the protease accelerates the hydrolysis of earthworm protein and promotes the action of staphylococcus carnosus. Some special proteins are degraded into functional polypeptides to form more unknown growth factors, so that the functions of inhibiting bacteria, promoting growth, attracting food, promoting digestion, enhancing immunity and the like of the earthworms are increased by multiple times, and the health-care and growth-promoting effects in production of broilers, laying hens, pigs and aquatic products are obvious.
The protein is decomposed thoroughly by fermentation, and the content of amino acid is increased. And the active ingredients and beneficial ingredients of the earthworms are not decomposed in the fermentation step and are retained in the fermented earthworm powder. Therefore, the earthworm polypeptide prepared by the preparation method has the advantages of full protein decomposition, basically existence in the form of amino acid, basically consistent protein content and amino acid content, and broad-spectrum antibacterial bacteriostasis.
On one hand, after the pH value is adjusted to 4.5-6.5 by adding phosphate into the earthworm fermentation liquor, the grinding process is in a stable state, the earthworm grinding liquor is ensured to be stable in shape in the grinding process, and the earthworm grinding liquor is not easy to deteriorate and stink in the later period. On the other hand, the protein is decomposed more thoroughly, so that the later period is not easy to deteriorate and smell.
Generally, the protein of the earthworm can reach 70 percent, and only supernatant fluid, namely a part with less protein content, can be selected under the condition that the earthworm is not fully decomposed in the conventional fermentation method, otherwise, the earthworm is deteriorated and smells. Therefore, the steps of filtration and the like are increased, and the yield of the earthworm powder is objectively reduced to a certain extent. The embodiment realizes full decomposition by fermenting the earthworm protein, does not deteriorate and smell, and has the condition of reserving and taking mixed liquid, so the steps are simplified, and the yield of the fermented earthworm powder is also improved. The earthworm fermentation liquor added with phosphate can be dried to be made into powder, so as to obtain fermented earthworm powder.
In the embodiment of the application, the earthworms are selected as raw materials, the earthworms are selected as animal protein feed raw materials, the earthworm feed contains unknown growth factors of animals, and the feeding attraction, the immunity enhancement, the body metabolism regulation and the intestinal health maintenance function of the earthworm feed are all superior to other animal protein raw materials (such as fish meal, meat meal, milk powder and the like).
In the embodiment of the application, functional proteins of earthworms (functional proteins for improving food calling, enhancing immunity, promoting body metabolism and maintaining intestinal health) are decomposed into small peptides with stronger functions, so that the functions of the earthworms for improving food calling, enhancing immunity, promoting body metabolism and maintaining intestinal health are amplified, and meanwhile, polypeptides (antibacterial peptides) with substitute-resistant functions are generated. The fermented earthworm powder has rich fermentation fragrance, obvious feeding attraction, and immune enhancing effect, body metabolism regulating effect and intestinal health maintaining effect superior to those of common animal protein material.
The fermented earthworm powder is fully decomposed to decompose protein into amino acid, and the decomposed protein is only 2-5% and the amino acid is low and is 2-10%. The protein content of the earthworm powder in the current market is high, and generally reaches more than 50%.
The fermented earthworm powder can be preserved at normal temperature; the earthworm active peptide is rich in special protein and functional polypeptide, contains unknown growth factors, contains 4530Da active peptide more than or equal to 115mg/kg, and shows that the active ingredients and beneficial ingredients of the earthworm are fully released; the product is high in temperature of 90 ℃, so that the activity of the effective components can be ensured to be stable, and the granulated feed for the broiler can be directly used; the fermented earthworm powder used by livestock and poultry can inhibit bacteria, promote growth, induce food, promote digestion and enhance immunity; the use of fermented earthworm powder in the compound feed for livestock and poultry can improve the utilization rate of crude protein by about 3 percent, the conversion rate of amino acid by about 5 percent, the utilization rate of calcium by about 6 percent, the utilization rate of phosphorus by about 4.5 percent and the energy utilization rate by about 1.5 percent.
According to the preparation method of the fermented earthworm powder, the protein is stably decomposed and the release of active ingredients and beneficial ingredients of the earthworms is promoted through proper grinding temperature and grinding time. During the fermentation process, the staphylococcus carnosus can decompose and convert earthworm self protein into peptide bond protein, and the smell of the fermented finished product is kept; the protease accelerates the hydrolysis of earthworm protein and promotes the action of staphylococcus carnosus. And the active ingredients and beneficial ingredients of the earthworms are not decomposed in the fermentation, so the fermented earthworm powder prepared by the preparation method of the fermented earthworm powder has low protein content, improved amino acid content and broad-spectrum antibacterial performance.
According to one embodiment of the application, the grinding temperature is gradually increased from 60 ℃ to 80 ℃ within 55-65 minutes, the grinding comprises a first grinding stage in the temperature rising process and a second grinding stage after the temperature rising process, and the total grinding time is 120 minutes.
The temperature of 60 ℃ is the temperature at which the earthworm protein can be decomposed, and the temperature of 80 ℃ is the temperature at which the earthworm protein is decomposed most quickly. Thus, the grinding initiation temperature can be selected from these two temperatures. Meanwhile, the temperature rise speed is also a factor causing protein decomposition and denaturation, the temperature rise time is controlled to be 55-65 minutes, the earthworm protein is stably decomposed, and the earthworm protein is not easy to denature. Namely, the grinding time in the first grinding stage is 55 to 65 minutes. The temperature rise time is preferably 60 minutes.
When the grinding temperature reaches 80 ℃, the second grinding stage is carried out, the second grinding time is not too long, otherwise, the second grinding time is possibly denatured, so that the total grinding time is controlled to be 120 minutes, and the earthworm protein is ensured to be thoroughly decomposed and have stable properties.
According to one embodiment of the application, the mixture comprises 68% by mass of staphylococcus carnosus and 32% by mass of protease.
Under the conditions, the effect of promoting the decomposition and the enzymolysis of the earthworm protein by fermentation is better, and the effect of improving the flavor of the earthworm product is better.
According to one embodiment of the application, the addition amount of the mixture is 15-25% of the mass of the earthworm grinding fluid.
Under the conditions, the addition amount of the mixture is proper, so that the reasonable fermentation speed and the good fermentation effect are ensured, and the waste of the microbial inoculum and the enzyme preparation is avoided.
According to one embodiment of the application, the phosphate is a mixture of dipotassium hydrogen phosphate and potassium dihydrogen phosphate or a mixture of disodium hydrogen phosphate and sodium dihydrogen phosphate.
And (3) obtaining earthworm fermentation liquor after the first fermentation is finished, adding a mixture of dipotassium hydrogen phosphate and potassium dihydrogen phosphate or a mixture of disodium hydrogen phosphate and sodium dihydrogen phosphate into the liquor, and performing functional release and activity stabilization operation at 78-82 ℃.
According to one embodiment of the application, the phosphate is a mixture of dipotassium hydrogen phosphate and potassium dihydrogen phosphate, the dosage of the dipotassium hydrogen phosphate is 3-6% of the mass of the earthworm fermentation liquor, and the dosage of the potassium dihydrogen phosphate is 1-3% of the mass of the earthworm fermentation liquor.
According to one embodiment of the present application, the phosphate is a mixture of disodium hydrogen phosphate and sodium dihydrogen phosphate, the amount of the disodium hydrogen phosphate is 3-6% of the mass of the earthworm fermentation broth, and the amount of the sodium dihydrogen phosphate is 1-3% of the mass of the earthworm fermentation broth.
According to one embodiment of the application, the water addition amount during grinding is 2-2.6 times of the mass of the earthworms. Under the condition, the concentration of the earthworm grinding fluid is proper, so that the subsequent fermentation process is facilitated.
According to one embodiment of the present application, the earthworm fermentation broth added with phosphate is made into powder by spray drying. Spray drying can retain unknown growth factors in the liquid to a greater extent.
According to one embodiment of the application, the step of preparing the powder from the earthworm fermentation liquor added with the phosphate by spray drying comprises the steps of mixing the earthworm fermentation liquor added with the phosphate with a carrier, and performing spray drying to prepare the powder.
The earthworm fermentation liquor is adsorbed by carriers such as wheat flour, starch, corn protein powder and the like for spray drying, so that uniform adsorption can be ensured.
The embodiment of the application also provides fermented earthworm powder prepared by the preparation method. The fermented earthworm powder prepared by the preparation method comprises 40 amino acid residues. The fermented earthworm powder has two obvious characteristics, namely, the earthworm powder has bacteriostasis (broad-spectrum antibiosis), and the earthworm powder has low protein content which is only 2-5 percent, the amino acid content is basically consistent with the protein content, and the sum of the 17 amino acid contents is 2-10 percent.
The fermented earthworm powder has rich fermentation fragrance, obvious food calling performance, and immunological enhancement, body metabolism regulation and intestinal health maintenance effects superior to those of common animal protein material.
The preparation method decomposes functional protein of Lumbricus into small peptide with stronger functionality, so as to enlarge functions of inducing food, enhancing immunity, promoting metabolism, and maintaining intestinal health, and generate polypeptide with substitute antibody function.
Example 1
1. Selecting fresh and live earthworms, and cleaning.
2. Grinding and physical crushing: grinding at 60-78 deg.C for 55 min from 60 deg.C to 78 deg.C, grinding at 80 deg.C for 85 min, and grinding for 140 min, wherein water is added during grinding period and the amount of water is 2.5 times of fresh Lumbricus.
3. Fermentation and decomposition: performing primary fermentation on the earthworm grinding liquid, adding a mixture with the mass of 15% of the filtrate during the fermentation, and performing fermentation for 7 days at the fermentation temperature of 80 ℃; the mixture comprises 66 mass percent of staphylococcus carnosus and 34 mass percent of protease.
4. And (3) stable release: after the fermentation is finished, earthworm fermentation liquor is obtained, dipotassium hydrogen phosphate with the dosage being 3% of the mass of the earthworm fermentation liquor is added into the earthworm fermentation liquor, and functional release and stable activity operation are carried out at 82 ℃.
5. Drying into powder: adding 10% of wheat flour as a carrier into the earthworm fermentation liquor added with phosphate, fully mixing and uniformly adsorbing, and then putting into a spray dryer for drying.
Example 2
1. Selecting fresh and live earthworms, and cleaning;
2. grinding earthworms: grinding at 62-80 deg.C for 65 min from 62 deg.C to 80 deg.C, setting grinding time at 82 deg.C for 35 min, wherein the total grinding time is 100 min, and adding water in the grinding period, wherein the water amount is 2.5 times of fresh Lumbricus amount;
3. fermentation and decomposition: carrying out primary fermentation on the earthworm grinding fluid, adding a mixture with the mass of 18% of the filtrate during fermentation, wherein the fermentation time is 7 days, and the fermentation temperature is 78 ℃; the mixture comprises 68 percent of staphylococcus carnosus and 32 percent of protease by mass.
4. And (3) stable release: after the fermentation is finished, earthworm fermentation liquor is obtained, disodium hydrogen phosphate with the dosage of 4% of the mass of the earthworm fermentation liquor is added into the earthworm fermentation liquor, and functional release and stable activity operation are carried out at 78 ℃.
5. Drying into powder: adding 13% of starch as a carrier into the earthworm fermentation liquor added with phosphate, fully mixing and uniformly adsorbing, and then putting into a spray dryer for drying.
Example 3
1. Selecting fresh and live earthworms, and cleaning;
2. grinding earthworms: grinding at 62-78 deg.C for 60 min from 62 deg.C to 80 deg.C, grinding at 80 deg.C for 70 min for 130 min, adding water 2.5 times of fresh earthworm;
3. fermentation and decomposition: carrying out primary fermentation on the earthworm grinding fluid, adding a mixture with the mass of 22% of the filtrate during fermentation, wherein the fermentation time is 7 days, and the fermentation temperature is 78 ℃; the mixture comprises 70 percent of staphylococcus carnosus and 30 percent of protease by mass percent.
4. And (3) stable release: after the fermentation is finished, earthworm fermentation liquor is obtained, monopotassium phosphate with the dosage being 1% of the mass of the earthworm fermentation liquor is added into the liquor, and functional release and stable activity operation are carried out at 82 ℃.
5. Drying into powder: adding 12% of corn protein powder as a carrier into the earthworm fermentation liquor added with phosphate, fully mixing and uniformly adsorbing, and then putting into a spray dryer for drying.
Example 4
1. Selecting fresh and live earthworms, and cleaning;
2. grinding earthworms: grinding at 60-80 deg.C for 60 min from 60 deg.C to 80 deg.C, setting grinding time at 80 deg.C for 60 min, wherein the total grinding time is 120 min, and adding water in the grinding period, the amount of water is 2.5 times of fresh earthworm;
3. fermentation and decomposition: carrying out primary fermentation on the earthworm grinding fluid, adding a mixture with the mass of 22% of the filtrate during fermentation, wherein the fermentation time is 7 days, and the fermentation temperature is 78 ℃; the mixture comprises 69 percent of staphylococcus carnosus and 31 percent of protease by mass.
4. And (3) stable release: after the fermentation is finished, earthworm fermentation liquor is obtained, sodium dihydrogen phosphate with the dosage being 3% of the mass of the earthworm fermentation liquor is added into the liquor, and functional release and stable activity operation are carried out at the temperature of 80 ℃.
5. Drying into powder: adding 15% of corn protein powder as a carrier into the earthworm fermentation liquor added with phosphate, fully mixing and uniformly adsorbing, and then putting into a spray dryer for drying.
Comparative example 1
Compared with example 4, step 2 is different, and the rest steps are the same.
In step 2, grinding earthworms: the grinding temperature is kept at 85 ℃, the grinding time is 120 minutes, and water is added during the grinding period, wherein the water addition amount is 2.5 times of that of the fresh earthworms.
Comparative example 2
Compared with example 4, step 2 is different, and the rest steps are the same.
In step 2, grinding earthworms: the grinding temperature is kept at 58 ℃, the grinding time is 120 minutes, and water is added during the grinding period, wherein the water addition amount is 2.5 times of that of the fresh earthworms.
Comparative example 3
Compared with example 4, step 3 is different, and the rest steps are the same.
In the step 3: performing primary fermentation on the earthworm grinding fluid, adding the mixture during the fermentation, wherein the fermentation time is 7 days, and the fermentation temperature is 80 ℃; the mixture comprises lactic acid bacteria, yeast and aspergillus, and the mass ratio of the lactic acid bacteria, the yeast and the aspergillus is 0.43:0.3: 0.25.
Comparative example 4
Compared with the embodiment 4, the step 4 is omitted, and the rest steps are the same.
And (3) taking the products of examples 1 to 4 and comparative examples 1 to 4 and commercial earthworm powder to carry out amino acid content detection, wherein the detection method of the amino acid is GB/T8246-2000. The products of examples 1 to 4 and comparative examples 1 to 4 and commercial earthworm powder were subjected to 4530Da active peptide content detection by electrophoresis. The results are shown in Table 1. The commercially available earthworm powder is produced by Xian Luteng biological technology Co., Ltd and has the model number lt 18060602.
TABLE 1 detection results of protein content and amino acid sum content
Figure BDA0002973859690000101
The detection result shows that the earthworm protein is decomposed into amino acid and peptide bond protein through full decomposition of the fermented earthworm powder, the content of the amino acid is less than 10 and is 2-10%, and the content of the active peptide is more than or equal to 115 mg/kg. The powder in comparative examples 1-4 has high amino acid content, low content of active peptide, and insufficient decomposition.
In addition, the earthworm fermentation broth added with phosphate in example 4, antibiotics, bacitracin zinc and a proper amount of deionized water were respectively taken to prepare solutions with different concentrations (see table 2), and the inhibition zones of staphylococcus aureus, bacillus subtilis, pseudomonas aeruginosa, enterococcus faecalis and escherichia coli were detected.
TABLE 2 gradient of solution configuration of substances
Figure BDA0002973859690000111
The results of the tests are shown in FIGS. 1 to 5 and Table 3.
TABLE 3 results of bacteriostatic tests
Figure BDA0002973859690000112
The detection result shows that:
1. the comparison shows that the earthworm fermentation liquor added with phosphate has more obvious bacteriostatic effect.
2. Compared with the 7-17 antibiotics-containing medicines, the earthworm fermentation liquor added with phosphate has the antibacterial effect obviously superior to antibiotics.
3. Through the comparison, part of the strains are low or insensitive to antibiotics, and the earthworm fermentation liquor added with phosphate has better or better inhibition effect on the strains.
The fermented earthworm powder prepared by the earthworm fermentation liquid added with phosphate also has broad-spectrum antibacterial performance.
While the present application has been described with reference to preferred embodiments, various modifications may be made and equivalents may be substituted for elements thereof without departing from the scope of the present application, and particularly, features described in connection with the embodiments may be combined in any manner as long as there is no structural conflict. The present application is not intended to be limited to the particular embodiments disclosed herein but is to cover all embodiments that may fall within the scope of the appended claims.

Claims (10)

1. A preparation method of fermented earthworm powder is characterized by comprising the following steps:
grinding earthworms for 100-140 minutes, gradually increasing the grinding temperature from 60-62 ℃ to 78-80 ℃, adding water during grinding, and obtaining earthworm grinding fluid after grinding;
adding a mixing agent into the earthworm grinding liquid, fermenting at 78-82 ℃, wherein the mixing agent comprises 66-70% of staphylococcus carnosus and 30-34% of neutral protease by mass, and performing fermentation and enzymolysis to obtain earthworm fermentation liquid;
adding phosphate into the earthworm fermentation liquor to adjust the pH value to 4.5-6.5, and keeping the earthworm fermentation liquor stable;
drying the earthworm fermentation liquor added with phosphate to prepare powder, thus obtaining the fermented earthworm powder.
2. The method for preparing fermented earthworm powder according to claim 1, wherein the grinding comprises a first grinding stage in a temperature rising process and a second grinding stage after the temperature rising process, the total grinding time is 120 minutes, the grinding temperature in the first grinding stage is gradually increased from 60 ℃ to 80 ℃, and the temperature rising time is 55-65 minutes.
3. The method for preparing fermented earthworm powder according to claim 2, wherein the mixture comprises 68% by mass of staphylococcus carnosus and 32% by mass of protease.
4. The method for preparing fermented earthworm powder according to claim 1, wherein the amount of the mixture is 15-25% by mass of the earthworm grinding fluid.
5. The method for producing fermented earthworm powder according to claim 1, wherein the phosphate is a mixture of dipotassium hydrogen phosphate and potassium dihydrogen phosphate or a mixture of disodium hydrogen phosphate and sodium dihydrogen phosphate.
6. The method for producing fermented earthworm powder according to claim 5, wherein the phosphate is a mixture of dipotassium hydrogen phosphate and potassium dihydrogen phosphate, the amount of the dipotassium hydrogen phosphate is 3 to 6% by mass of the earthworm fermentation solution, and the amount of the potassium dihydrogen phosphate is 1 to 3% by mass of the earthworm fermentation solution.
7. The method for producing fermented earthworm powder according to claim 5, wherein the phosphate is a mixture of disodium hydrogen phosphate and sodium dihydrogen phosphate, the amount of disodium hydrogen phosphate is 3 to 6% by mass of the earthworm fermentation broth, and the amount of sodium dihydrogen phosphate is 1 to 3% by mass of the earthworm fermentation broth.
8. The method for preparing fermented earthworm powder according to claim 2, wherein water is added twice during the grinding period, and the total added water amount is 2-2.6 times of the mass of the earthworms.
9. The method for producing fermented earthworm powder according to claim 1, wherein the earthworm fermentation broth added with phosphate is mixed with a carrier and spray-dried to produce a powder, and the amount of the carrier added is 10% or more of the mass of the earthworm fermentation broth added with phosphate.
10. Fermented earthworm powder prepared by the preparation method according to any one of claims 1-9.
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CN104593456A (en) * 2014-12-02 2015-05-06 高向阳 Method for preparing peptone from enzymatically-hydrolyzed earthworm dry powder and purpose of peptone

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GB0202556D0 (en) * 2002-02-04 2002-03-20 Danisco Novel Protein
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CN102160640A (en) * 2011-02-18 2011-08-24 郑鉴忠 Earthworm diet therapy nutritious food production method
CN104593456A (en) * 2014-12-02 2015-05-06 高向阳 Method for preparing peptone from enzymatically-hydrolyzed earthworm dry powder and purpose of peptone

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