CN114190561A - Microencapsulated royal jelly enzymolysis polypeptide, preparation method thereof and solid powder containing microencapsulated royal jelly enzymolysis polypeptide - Google Patents
Microencapsulated royal jelly enzymolysis polypeptide, preparation method thereof and solid powder containing microencapsulated royal jelly enzymolysis polypeptide Download PDFInfo
- Publication number
- CN114190561A CN114190561A CN202111499532.0A CN202111499532A CN114190561A CN 114190561 A CN114190561 A CN 114190561A CN 202111499532 A CN202111499532 A CN 202111499532A CN 114190561 A CN114190561 A CN 114190561A
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- CN
- China
- Prior art keywords
- royal jelly
- microencapsulated
- weight
- enzymolysis
- polypeptide
- Prior art date
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- Pending
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Abstract
The application relates to a microencapsulated royal jelly enzymatic hydrolysis polypeptide, a preparation method thereof and solid powder containing the same. The preparation method of the microencapsulated royal jelly enzymatic hydrolysis polypeptide comprises the following steps: (1) mixing royal jelly with water, and centrifuging to obtain supernatant; (2) adding pepsin into the supernatant obtained in the step (1) for enzymolysis to obtain primary enzymolysis liquid; (3) adding trypsin into the primary enzymolysis liquid for enzymolysis to obtain secondary enzymolysis liquid; (4) centrifuging the secondary enzymolysis liquid, and taking supernatant; (5) and (4) adding an embedding wall material into the supernatant obtained in the step (4), and grinding and homogenizing. The preparation method can fully obtain the active ingredients of the royal jelly, improve the taste of the royal jelly, form microencapsulation and embed the royal jelly zymolytic polypeptide to the maximum extent. In addition, the solid powder contains microencapsulated royal jelly enzymatic hydrolysis polypeptide and queen bee larva enzymatic hydrolysis polypeptide, and has good functions of relieving fatigue, enhancing physical strength and the like.
Description
The technical field is as follows:
the application belongs to the field of edible and medicinal health care products, and particularly relates to microencapsulated royal jelly enzymatic hydrolysis polypeptide, a preparation method thereof and solid powder containing the same.
Background art:
royal Jelly (Royal Jelly), also known as Royal Jelly, Royal Jelly and the like, is pulpy substances secreted by lingual glands (pharyngeal glands) and palatal glands of worker bees of 5-15 days old, is milk-white or light yellow, is a special pulpy substance specially used for feeding queen bees and young bees of 3 days old, is unique food for the worker bees and the male peak larvae in the whole development period and within 3 days old of the queen bee larvae, is also food for the life of the queen bees, is similar to the milk of mammals, and is also called Royal Jelly. The royal jelly contains complex bioactive substances, and has high nutritive value and health promotion function. The fresh royal jelly contains 11-14% of protein and 38-50% of royal jelly dry matter, and the protein has the highest proportion in dry matter, and is rich in amino acids, lipids, vitamins and minerals. Numerous studies and clinical applications suggest: the royal jelly has the functions of regulating immunity, resisting fatigue, resisting bacteria, diminishing inflammation, resisting allergy, resisting aging, resisting cancer, resisting tumor, expanding blood vessels, reducing blood pressure, reducing blood sugar, resisting high cholesterol, resisting oxidation, improving sleep, promoting cell growth, promoting insulin-like activity and the like, and is widely applied to the fields of food, health care products, medicines, cosmetics and the like.
Among the numerous nutrition and health care functions of royal jelly, the anti-fatigue effect is not widely regarded. Fatigue is the symptom of decreased muscle strength caused by a series of biochemical changes in the body caused by exercise. There are two main methods for evaluating fatigue: exercise endurance testing and detection of biochemical changes. Exercise endurance is the most direct and objective index reflecting body fatigue. As can be seen from the experiment of swimming with the weight load of minus 5 percent at the tail of a mouse, the exercise endurance of the fresh royal jelly, the filter residue components filtered by a 180-mesh filter screen and the filter residue components filtered by a 0.5-micron inorganic ceramic membrane of the filtrate can be improved, and the royal jelly is proved to have a certain anti-fatigue effect.
China is the country which produces and outputs royal jelly most in the world, most royal jelly products are raw material products, however, the royal jelly is spicy, viscous and poor in solubility, and the sensory and physicochemical properties restrict the market space and the economic value of the royal jelly. The royal jelly is rich in protein, and the protein content of the royal jelly accounts for about 30-50% of dry matters of the royal jelly. The important biological functions and medical care functions of the royal jelly are probably closely related to the main proteins (MRJPs) of the royal jelly. Currently, 9 members of the MRJPs family have been identified: MRJPs1-9, the molecular mass of which is in the range of 49-87 ku. In terms of biological functions, MRJPs provide nutrients for the growth, development and reproduction of bees. In the aspect of health care efficacy, MRJPs mainly have the effects of promoting cell proliferation, high-efficiency antibacterial activity, strong immunological activity, antioxidant activity, anti-fatigue and the like. The research shows that MRJP 1 can stimulate the growth of human lymphocytes in a serum-free culture medium, can also promote the proliferation capacity of mouse hepatocytes and shows a certain anti-fatigue effect. From the nutrition perspective, the use value of the protein of the royal jelly is also severely restricted by taking the royal jelly as a raw material product, most of the protein extracted and separated from the royal jelly is large molecular weight protein which can not be directly absorbed by a human body, and the action mechanism in the human body has certain ambiguity.
In addition, queen bee larva, also called queen bee larva, queen bee larva and queen bee larva, is an embryo nutrient body developed by sucking royal jelly before bee eggs hatch to larva become pupae, has rich and comprehensive nutrient components, and is a natural high-grade nutrient product with high protein, low fat, multiple vitamins and low sugar. According to the reports at home and abroad, the queen bee larva not only can stimulate appetite, improve sleep and build up health of weak patients, but also can regulate the central nervous system, endocrine metabolism and the like of human bodies. Is suitable for patients with asthenia, fatigue, asthenia, malnutrition, postoperative patients, old people, and people in need of nourishing and rejuvenating, and has adjuvant therapeutic effect on neurasthenia, rheumatic arthritis, liver disease and leukopenia. The queen bee larva is rich in nutrition and active ingredients, which goes without saying, but is easy to be oxidized and deteriorated, so that the processing, storage and utilization of the queen bee larva are limited.
Therefore, how to increase the absorption rate and the efficacy of the royal jelly, reduce the occurrence of anaphylactic reaction and how to develop the anti-fatigue efficacy of the bee product has important significance.
The invention content is as follows:
in view of the above-mentioned deficiencies of the prior art, an object of the present application is to provide a method for preparing microencapsulated royal jelly enzymatic hydrolysis polypeptide, which can fully obtain the active ingredients of royal jelly, improve the use value of royal jelly, improve the taste of royal jelly, and form microencapsulation, thereby embedding royal jelly enzymatic hydrolysis polypeptide to the maximum extent, reducing the hygroscopicity of microcapsule wall material, and improving the stability of wall material.
Another objective of the present application is to provide a microencapsulated royal jelly enzymatic polypeptide prepared by the above preparation method, wherein the royal jelly enzymatic polypeptide contains a large amount of active polypeptides, which can effectively improve the biological activity, increase the absorption rate and reduce the allergic reaction, and the microencapsulated structure also provides excellent stability.
Still another object of this application is to provide a solid powder containing microencapsulated royal jelly enzymatic polypeptide, which further contains queen bee larva enzymatic polypeptide (bee larva peptide). The bee larva is subjected to enzymolysis to prepare the bee larva peptide, and the bee larva peptide is scientifically compatible with the microencapsulated bee royal jelly enzymolysis polypeptide, so that the stability of the bee larva peptide is effectively improved, and meanwhile, the solid powder has good functions of relieving fatigue, enhancing physical strength and the like.
Still another object of the present application is to provide a method for preparing the solid powder containing the microencapsulated royal jelly enzymatic hydrolysis polypeptide, wherein the preparation method is simple, convenient and fast to operate and has high efficiency.
In order to achieve the above objects, in a first aspect, the present application provides a method for preparing a microencapsulated royal jelly enzymatic polypeptide, comprising the following steps:
(1) mixing royal jelly with water, and centrifuging to obtain supernatant;
(2) adding pepsin into the supernatant obtained in the step (1) for enzymolysis to obtain primary enzymolysis liquid;
(3) adding trypsin into the primary enzymolysis liquid for enzymolysis to obtain secondary enzymolysis liquid;
(4) centrifuging the secondary enzymolysis liquid, and taking supernatant; and
(5) and (4) adding an embedding wall material into the supernatant obtained in the step (4), and grinding and homogenizing to obtain the microencapsulated royal jelly enzymatic hydrolysis polypeptide.
In one possible embodiment in combination with the first aspect, in the step (1), the weight ratio of the royal jelly to the water may be 1 (0.8-1).
In a possible embodiment in combination with the first aspect, the mixing in step (1) is performed under ultrasonic conditions.
With reference to the first aspect, in a possible embodiment, the enzymolysis in step (2) may be: enzymolysis is carried out for 40min to 60min under the temperature condition of 38 ℃ to 45 ℃.
In a possible embodiment in combination with the first aspect, in the step (2), the enzyme substrate concentration ratio may be 0.3% to 0.5%.
With reference to the first aspect, in a possible embodiment, the enzymolysis in step (3) is: adjusting the pH value of the system to 8, adding trypsin at the temperature of 38-45 ℃ and carrying out enzymolysis for 40-60 min.
Further, in the step (3), a sodium hydroxide solution with a mass concentration of 5% to 8% is used to adjust the pH.
In a possible embodiment in combination with the first aspect, in the step (3), the enzyme substrate concentration ratio may be 0.5% to 0.7%.
In a possible embodiment in combination with the first aspect, in the step (5), the embedding wall material is added in an amount of 0.5 to 10% by mass/volume (w/v).
Further, the embedding wall material comprises sodium carboxymethyl cellulose and beta-cyclodextrin in a mass ratio of 1 (0.1-3).
In a possible embodiment in combination with the first aspect, in the step (5), the time for grinding homogenization is 2 to 8 hours.
In a possible embodiment in combination with the first aspect, the step (5) further comprises concentrating the obtained material under vacuum at 40-50 ℃ after grinding homogenization is completed, and then spray drying, thereby obtaining the microencapsulated royal jelly enzymatic polypeptide.
In a second aspect, the application provides a microencapsulated royal jelly enzymatic polypeptide prepared by the preparation method.
In a third aspect, the present application provides a solid powder comprising microencapsulated royal jelly enzymatic polypeptide, which comprises:
in one possible embodiment in combination with the third aspect, the melittin is prepared by:
pulverizing queen bee embryo, mixing with water to obtain slurry, and adding alkaline protease for enzymolysis to obtain foetus Apis peptide.
Further, the queen bee larva is sieved by a 30-mesh sieve after being crushed.
Further, queen bee larva powder and water are mixed according to the mass ratio of 1 (8-12), preferably 1: 10.
Further, the ratio of the concentration of the alkaline protease to the substrate may be 1% to 3%, preferably 2%.
Further, the enzymolysis is carried out for 2 to 6 hours at the temperature of between 40 and 50 ℃.
With reference to the third aspect, in one possible embodiment, the solid powder further includes:
0.2 to 0.4 part by weight of D-mannitol, and
0.1-0.2 parts of nobiletin.
Further, the mass ratio of the D-mannitol to the nobiletin is 2: 1.
In a fourth aspect, the application provides a preparation method of the solid powder containing the microencapsulated royal jelly enzymatic hydrolysis polypeptide, which comprises the following steps:
and uniformly mixing the components to obtain the solid powder containing the microencapsulated royal jelly enzymatic hydrolysis polypeptide.
According to the technical scheme provided by the application, compared with the prior art, the method at least comprises the following beneficial effects:
according to the preparation method of the microencapsulated royal jelly enzymatic hydrolysis polypeptide, the active ingredients of the royal jelly can be fully obtained, the use value of the royal jelly is improved, the mouthfeel of the royal jelly is improved, in addition, the microencapsulated royal jelly enzymatic hydrolysis polypeptide is formed, the royal jelly enzymatic hydrolysis polypeptide is embedded to the maximum extent, the hygroscopicity of a microcapsule wall material is reduced, and the stability of the wall material is improved. The microencapsulated royal jelly enzymatic polypeptide prepared by the preparation method contains a large amount of active polypeptide, can effectively improve the biological activity of the polypeptide, also increases the absorption rate and reduces the anaphylactic reaction, and meanwhile, the microencapsulated structure also ensures that the polypeptide has excellent stability. In addition, the solid powder comprising the microencapsulated royal jelly enzymatic polypeptide according to the present application further comprises queen bee larva enzymatic polypeptide (bee larva peptide). The bee larva is subjected to enzymolysis to prepare the bee larva peptide, and the bee larva peptide is scientifically compatible with the microencapsulated bee royal jelly enzymolysis polypeptide, so that the stability of the bee larva peptide is effectively improved, and meanwhile, the solid powder has good functions of relieving fatigue, enhancing physical strength and the like. The preparation method of the solid powder is simple, convenient and quick, and high in efficiency.
Detailed Description
In order that those skilled in the art will be able to more clearly understand the present application, the present application will be described in detail below with reference to examples. Before the description is given, it should be understood that the terms used in the present specification and the appended claims should not be construed as limited to general and dictionary meanings, but interpreted based on the meanings and concepts corresponding to technical aspects of the present application on the basis of the principle that the inventor is allowed to define terms appropriately for the best explanation. Accordingly, the description proposed herein is just a preferable example for the purpose of illustrations only, not intended to limit the scope of the application, so it should be understood that other equivalents and modifications could be made thereto without departing from the spirit and scope of the application, and the scope of the application claims should be determined only by the claims. Unless otherwise specified, reagents and equipment used in the following examples are commercially available products.
The method is realized by the following technical scheme:
in a first aspect, the application provides a preparation method of a microencapsulated royal jelly enzymatic polypeptide, which comprises the following steps:
(1) mixing royal jelly with water, and centrifuging to obtain supernatant;
(2) adding pepsin into the supernatant obtained in the step (1) for enzymolysis to obtain primary enzymolysis liquid;
(3) adding trypsin into the primary enzymolysis liquid for enzymolysis to obtain secondary enzymolysis liquid;
(4) centrifuging the secondary enzymolysis liquid, and taking supernatant; and
(5) and (4) adding an embedding wall material into the supernatant obtained in the step (4), and grinding and homogenizing to obtain the microencapsulated royal jelly enzymatic hydrolysis polypeptide.
In the application, the protein in the royal jelly is decomposed into small molecular peptide fragments by a proteolysis technology, so that the problem of the allergy of the royal jelly can be well solved, and the biological activity of the royal jelly can be further effectively improved. The bioactive peptide obtained by enzymolysis has the special functions of eliminating free radicals, improving the immunity of an organism, delaying senescence, reducing blood pressure and the like, cannot show activity in an original protein sequence, and is released from the original sequence under the action of intestinal enzymes or in vitro enzymes of a human body and shows special activity. According to the method, the royal jelly enzymolysis polypeptide is prepared by adopting two-stage enzymolysis of pepsin and trypsin, so that active ingredients in the royal jelly can be more fully obtained, the use value of the royal jelly is greatly improved, and the taste of the royal jelly is greatly improved.
In addition, the prepared royal jelly enzymolysis polypeptide is embedded by adding the embedding wall material, so that the polypeptide can be well stabilized, various biological activities of the polypeptide can be effectively maintained, the shelf life can be prolonged, and poor taste can be further covered.
In a possible embodiment, in combination with the first aspect, in step (1), the weight ratio of royal jelly to water may be 1 (0.8-1), for example, 1:0.8, 1:0.85, 1:0.9, 1:0.95 or 1:1, or other ratio values within the range). Within this ratio range, the royal jelly can be effectively diluted, thereby facilitating the subsequent centrifugation process. If the water consumption is too low, it is difficult to perform a centrifugal separation process to remove unnecessary impurities, and if the water consumption is too high, the concentration of effective components is too low, which is not favorable for post-treatment.
In a possible embodiment in combination with the first aspect, the mixing in step (1) is performed under ultrasonic conditions. Through the supersound, can be swift effectively with royal jelly and water misce bene.
With reference to the first aspect, in a possible embodiment, the enzymolysis in step (2) may be: enzymolysis is carried out for 40min to 60min (for example, 40min, 42min, 44min, 46min, 48min, 50min, 52min, 54min, 56min, 58min or 60min, or other specific time value in the range) under the temperature condition of 38 ℃ to 45 ℃ (for example, 38 ℃, 39 ℃, 40 ℃, 41 ℃, 42 ℃, 43 ℃, 44 ℃ or 45 ℃, or other specific temperature value in the range). Under the condition of proper temperature and time, the royal jelly can be effectively enzymolyzed to obtain primary enzymolysis liquid.
In one possible embodiment in combination with the first aspect, in step (2), the enzyme substrate concentration ratio may be 0.3% to 0.5%, for example, may be 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, or 0.5%, or other specific percentage value within the range. By using the enzyme substrate concentration within the above-mentioned suitable range, neither the concentration is too low to allow efficient enzymatic hydrolysis, nor is the concentration too high to increase costs and cause excessive enzymatic hydrolysis.
With reference to the first aspect, in a possible embodiment, the enzymolysis in step (3) is: adjusting the pH of the system to 8, adding trypsin at 38-45 deg.C (for example, 38 deg.C, 39 deg.C, 40 deg.C, 41 deg.C, 42 deg.C, 43 deg.C, 44 deg.C or 45 deg.C, or other specific temperature value in the range), and performing enzymolysis for 40-60 min (for example, 40min, 42min, 44min, 46min, 48min, 50min, 52min, 54min, 56min, 58min or 60min, or other specific time value in the range). Under the condition of proper temperature and time, the primary enzymatic hydrolysate of the royal jelly can be continuously and effectively subjected to enzymolysis to obtain secondary enzymatic hydrolysate.
Further, in the step (3), a sodium hydroxide solution with a mass concentration of 5% to 8% (for example, 5%, 6%, 7%, or 8%, or other specific percentage value within the range) is used to adjust the pH. With sodium hydroxide solutions in this concentration range, the pH of the system can be adjusted quickly in small amounts, neither slowly due to too low a concentration, nor easily in excess alkalinity due to too high a concentration.
In one possible embodiment in combination with the first aspect, in said step (3), the enzyme substrate concentration ratio may be 0.5% to 0.7%, for example, may be 0.5%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0.59%, 0.6%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, or 0.7%, or other specific percentage value within said range. By using the enzyme substrate concentration within the above-mentioned suitable range, neither the concentration is too low to allow efficient enzymatic hydrolysis, nor is the concentration too high to increase costs and cause excessive enzymatic hydrolysis.
Through the two-stage enzymolysis including the primary enzymolysis and the secondary enzymolysis, the royal jelly can be more fully enzymolyzed to obtain active polypeptide components in the royal jelly, and the taste of the royal jelly is greatly improved.
In a possible embodiment in combination with the first aspect, in the step (5), the embedded wall material is added in an amount of 0.5 to 10% by mass/volume (w/v), for example, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, or 10%, or other specific percentage value within the range. And (4) the mass-to-volume ratio is the ratio of the mass of the embedding wall material to the volume of the supernatant obtained in the step (4). Under the use amount of the embedding wall materials in the mass-volume ratio, microencapsulation can be well formed, and the royal jelly enzymolysis polypeptide is coated to the maximum extent, so that the stability of the royal jelly enzymolysis polypeptide is improved, the shelf life is prolonged, and the stimulating taste can be covered. If the mass-to-volume ratio is too low, the microcapsule cannot be sufficiently microencapsulated, and if the mass-to-volume ratio is too high, the royal jelly enzymatic hydrolysis polypeptide is excessively wrapped, so that the royal jelly enzymatic hydrolysis polypeptide is not easily and sufficiently released and absorbed after being taken, and the cost is also increased.
Further, the embedding wall material comprises sodium carboxymethyl cellulose and beta-cyclodextrin in a mass ratio of 1 (0.1-3). In the present application, the mass ratio of sodium carboxymethylcellulose to β -cyclodextrin may be 1 (0.1 to 3), for example, 1:0.1, 1:0.2, 1:0.3, 1:0.4, 1:0.5, 1:0.7, 1:0.9, 1:1, 1:1.5, 1:2, 1:2.5, or 1:3, or other specific ratios within the range. Within the range of the ratio, the royal jelly enzymolysis polypeptide can be effectively microencapsulated in various combination forms, so that the advantages of beta-cyclodextrin and carboxymethyl cellulose (CMC) are complemented, the royal jelly polypeptide is embedded to the maximum extent, the hygroscopicity of the wall material is reduced, and the stability of the wall material is improved.
In a possible embodiment in combination with the first aspect, in step (5), the time for milling homogenization is 2h to 8h, for example, 2h, 2.5h, 3h, 3.5h, 4h, 4.5h, 5h, 5.5h, 6h, 6.5h, 7h, 7.5h, or 8h, or other specific time values within the range. Within this range, the encapsulating wall material can be sufficiently dispersed in the supernatant obtained in step (4) to achieve microencapsulation.
In a possible embodiment in combination with the first aspect, the step (5) further includes vacuum concentrating the obtained substance at 40-50 ℃ (for example, 40 ℃, 41 ℃, 42 ℃, 43 ℃, 44 ℃, 45 ℃, 46 ℃, 47 ℃, 48 ℃, 49 ℃ or 50 ℃, or other specific temperature value in the range) after grinding homogenization is completed, and then spray drying, thereby obtaining the microencapsulated royal jelly enzymatic polypeptide. By concentrating and drying, a solid product can be obtained, and is convenient to store and subpackage.
In a second aspect, the application provides a microencapsulated royal jelly enzymatic polypeptide prepared by the preparation method. The royal jelly enzymolysis polypeptide contains a large amount of active polypeptide, can effectively improve the biological activity of the royal jelly enzymolysis polypeptide, also increases the absorption rate, reduces the anaphylactic reaction, and simultaneously has excellent stability due to the microencapsulation structure.
In a third aspect, the present application provides a solid powder comprising microencapsulated royal jelly enzymatic polypeptide, which comprises:
the solid powder is prepared by scientifically matching the bee larva peptide prepared by enzymolysis of the queen bee larva with the microencapsulated royal jelly enzymolysis polypeptide, so that the stability of the bee larva peptide is effectively improved, and meanwhile, the solid powder has good functions of relieving fatigue, enhancing physical strength and the like.
In the present application, according to a possible embodiment, the microencapsulated royal jelly enzymatic polypeptide may be contained in the solid powder in an amount of any specific value within a range of 100 to 120 parts by weight, for example, 100 parts by weight, 101 parts by weight, 102 parts by weight, 103 parts by weight, 104 parts by weight, 105 parts by weight, 106 parts by weight, 107 parts by weight, 108 parts by weight, 109 parts by weight, 110 parts by weight, 111 parts by weight, 112 parts by weight, 113 parts by weight, 114 parts by weight, 115 parts by weight, 116 parts by weight, 117 parts by weight, 118 parts by weight, 119 parts by weight, or 120 parts by weight, or other specific values within the range; the content of the melittin may be any specific value within the range of 60 to 80 parts by weight, for example, 60 parts by weight, 61 parts by weight, 62 parts by weight, 63 parts by weight, 64 parts by weight, 65 parts by weight, 66 parts by weight, 67 parts by weight, 68 parts by weight, 69 parts by weight, 70 parts by weight, 71 parts by weight, 72 parts by weight, 73 parts by weight, 74 parts by weight, 75 parts by weight, 76 parts by weight, 77 parts by weight, 78 parts by weight, 79 parts by weight, or 80 parts by weight, or other specific values within the range; the maltodextrin may be present in an amount of any specific value within the range of 20 to 35 parts by weight, for example, 20 parts by weight, 21 parts by weight, 22 parts by weight, 23 parts by weight, 24 parts by weight, 25 parts by weight, 26 parts by weight, 27 parts by weight, 28 parts by weight, 29 parts by weight, 30 parts by weight, 31 parts by weight, 32 parts by weight, 33 parts by weight, 34 parts by weight, or 35 parts by weight, or other specific values within the range; and the isomalt content may be any specific value within the range of 10 to 15 parts by weight, for example, may be 10 parts by weight, 11 parts by weight, 12 parts by weight, 13 parts by weight, 14 parts by weight, or 15 parts by weight, or other specific values within the range. Within the composition and content range, the microencapsulated royal jelly enzymatic polypeptide can be effectively matched with the bee embryo peptide, so that the effects of relieving fatigue, enhancing physical strength and the like are exerted to the maximum extent.
In one possible embodiment in combination with the third aspect, the melittin is prepared by: pulverizing queen bee embryo, mixing with water to obtain slurry, and adding alkaline protease for enzymolysis to obtain foetus Apis peptide. By the enzymolysis of the alkaline protease, the protein in the queen bee can be converted into the nutritional polypeptide which is easy to be absorbed and utilized by human bodies, and the effect of the nutritional polypeptide is fully exerted.
Further, the queen bee larva is sieved by a 30-mesh sieve after being crushed. By sieving, larger particulate impurities can be removed.
Further, queen bee larva powder and water are mixed according to the mass ratio of 1 (8-12), for example, the mass ratio can be 1:8, 1:9, 1:10, 1:11 or 1:12, or other specific ratios in the range. Within this range, it is helpful to obtain a queen bee embryo slurry suitable for enzymatic hydrolysis. Preferably, the mass ratio may be 1: 10.
Further, the ratio of alkaline protease to substrate concentration may be 1% to 3%, for example, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%, 2.2%, 2.4%, 2.6%, 2.8%, or 3%, or other specific percentage values within the stated range. By using the enzyme substrate concentration within the above-mentioned suitable range, neither the concentration is too low to allow efficient enzymatic hydrolysis, nor is the concentration too high to increase costs and cause excessive enzymatic hydrolysis. Preferably, the ratio of the concentration of alkaline protease to substrate may be 2%.
Further, the enzymatic hydrolysis is performed for 2h to 6h (e.g., 2h, 2.5h, 3h, 3.5h, 4h, 4.5h, 5h, 5.5h, or 6h, or other specific time values within the range) under a temperature condition of 40 ℃ to 50 ℃ (e.g., 40 ℃, 41 ℃, 42 ℃, 43 ℃, 44 ℃, 45 ℃, 46 ℃, 47 ℃, 48 ℃, 49 ℃, or 50 ℃, or other specific temperature values within the range). By the above conditions, effective enzymatic hydrolysis of the melittin can be obtained.
With reference to the third aspect, in one possible embodiment, the solid powder further includes:
0.2 to 0.4 part by weight of D-mannitol, and
0.1-0.1 part by weight of nobiletin.
In the present application, according to one possible embodiment, the content of D-mannitol in the solid powder may be any specific value within the range of 0.2 to 0.4 parts by weight, for example, may be 0.2 parts by weight, 0.21 parts by weight, 0.22 parts by weight, 0.23 parts by weight, 0.24 parts by weight, 0.25 parts by weight, 0.26 parts by weight, 0.27 parts by weight, 0.28 parts by weight, 0.29 parts by weight, 0.3 parts by weight, 0.31 parts by weight, 0.32 parts by weight, 0.33 parts by weight, 0.34 parts by weight, 0.35 parts by weight, 0.36 parts by weight, 0.37 parts by weight, 0.38 parts by weight, 0.39 parts by weight, or 0.4 parts by weight, or other specific values within the range; and the content of nobiletin may be any specific value within the range of 0.1 to 0.2 parts by weight, for example, may be 0.1 part by weight, 0.11 part by weight, 0.12 part by weight, 0.13 part by weight, 0.14 part by weight, 0.15 part by weight, 0.16 part by weight, 0.17 part by weight, 0.18 part by weight, 0.19 part by weight, or 0.2 part by weight, or other specific values within the range. By adding D-mannitol and nobiletin with specific contents, the effects of relieving fatigue, enhancing physical strength and the like of the solid powder can be further synergistically improved. The inventors have found that the improvement is limited to the above content range, and that when the content of D-mannitol and nobiletin is too high or too low, the corresponding effect cannot be obtained, and the opposite may be obtained.
Further, the mass ratio of the D-mannitol to the nobiletin is 2: 1. Under the mass ratio, the effect of improving the efficacy of the D-mannitol and nobiletin on the solid powder can be more exhibited.
In a fourth aspect, the application provides a preparation method of the solid powder containing the microencapsulated royal jelly enzymatic hydrolysis polypeptide, which comprises the following steps: and uniformly mixing the components to obtain the solid powder containing the microencapsulated royal jelly enzymatic hydrolysis polypeptide. The preparation method is simple, convenient and quick to operate and high in efficiency.
According to the preparation method of the microencapsulated royal jelly enzymatic hydrolysis polypeptide, the protein in the royal jelly is decomposed into small molecular peptide fragments by a proteolysis technology, so that the active ingredients of the royal jelly can be fully obtained, the problem of anaphylaxis of the royal jelly is solved, the use value of the royal jelly is improved, the taste of the royal jelly is improved, microencapsulation is formed, the royal jelly enzymatic hydrolysis polypeptide is embedded to the maximum extent, various biological activities of the royal jelly are effectively maintained, the quality guarantee period is prolonged, the hygroscopicity of microcapsule wall materials is reduced, and the stability of the wall materials is improved. The microencapsulated royal jelly enzymatic polypeptide prepared by the preparation method contains a large amount of active polypeptide, can effectively improve the biological activity of the polypeptide, also increases the absorption rate and reduces the anaphylactic reaction, and meanwhile, the microencapsulated structure also ensures that the polypeptide has excellent stability. In addition, the solid powder comprising the microencapsulated royal jelly enzymatic polypeptide according to the present application further comprises queen bee larva enzymatic polypeptide (bee larva peptide). The bee larva is subjected to enzymolysis to prepare the bee larva peptide, and the bee larva peptide is scientifically compatible with the microencapsulated bee royal jelly enzymolysis polypeptide, so that the stability of the bee larva peptide is effectively improved, and meanwhile, the solid powder has good functions of relieving fatigue, enhancing physical strength and the like. Moreover, the D-mannitol and the nobiletin with specific contents are added into the solid powder, so that the efficacy of the solid powder can be further improved synergistically. The preparation method of the solid powder is simple, convenient and quick, and high in efficiency.
Examples
Example 1
The microencapsulated royal jelly enzymatic polypeptide is prepared by adopting the following preparation method of the microencapsulated royal jelly enzymatic polypeptide according to the application:
(1) mixing royal jelly and water in a weight ratio of 1:0.8 under an ultrasonic condition, and centrifuging to obtain supernatant;
(2) adding pepsin into the supernatant obtained in the step (1) according to the concentration ratio of 0.3% of enzyme substrate, and carrying out enzymolysis for 50min at the temperature of 45 ℃ to obtain primary enzymolysis liquid;
(3) adjusting the pH of the primary enzymolysis liquid to 8 by adopting a sodium hydroxide solution with the mass concentration of 5%, adding trypsin according to the enzyme substrate concentration ratio of 0.7% at the temperature of 40 ℃, and carrying out enzymolysis for 40min to obtain a secondary enzymolysis liquid;
(4) centrifuging the secondary enzymolysis liquid, and taking supernatant;
(5) and (3) adding an embedding wall material (sodium carboxymethylcellulose and beta-cyclodextrin in a mass ratio of 1: 1) into the supernatant obtained in the step (4) in a mass volume ratio of 2%, grinding and homogenizing for 4h, then carrying out vacuum concentration at 45 ℃, and then carrying out spray drying to obtain the microencapsulated royal jelly enzymatic hydrolysis polypeptide according to the application.
Example 2
The microencapsulated royal jelly enzymatic polypeptide is prepared by adopting the following preparation method of the microencapsulated royal jelly enzymatic polypeptide according to the application:
(1) mixing royal jelly and water in a weight ratio of 1:1 under an ultrasonic condition, and centrifuging to obtain supernatant;
(2) adding pepsin into the supernatant obtained in the step (1) according to the concentration ratio of 0.4% of enzyme substrate, and carrying out enzymolysis for 40min at the temperature of 40 ℃ to obtain primary enzymolysis liquid;
(3) adjusting the pH of the primary enzymolysis liquid to 8 by adopting a sodium hydroxide solution with the mass concentration of 8%, adding trypsin according to the concentration ratio of 0.6% of enzyme substrate at the temperature of 45 ℃, and carrying out enzymolysis for 60min to obtain a secondary enzymolysis liquid;
(4) centrifuging the secondary enzymolysis liquid, and taking supernatant;
(5) and (3) adding an embedding wall material (sodium carboxymethyl cellulose and beta-cyclodextrin with the mass ratio of 1: 0.1) into the supernatant obtained in the step (4) in a mass volume ratio of 0.5%, grinding and homogenizing for 6 hours, then carrying out vacuum concentration at 50 ℃, and then carrying out spray drying to obtain the microencapsulated royal jelly enzymatic hydrolysis polypeptide according to the application.
Example 3
The microencapsulated royal jelly enzymatic polypeptide is prepared by adopting the following preparation method of the microencapsulated royal jelly enzymatic polypeptide according to the application:
(1) mixing royal jelly and water in a weight ratio of 1:0.9 under ultrasonic condition, and centrifuging to obtain supernatant;
(2) adding pepsin into the supernatant obtained in the step (1) according to the concentration ratio of 0.5% of enzyme substrate, and carrying out enzymolysis for 60min at the temperature of 38 ℃ to obtain primary enzymolysis liquid;
(3) adjusting the pH of the primary enzymolysis liquid to 8 by adopting a sodium hydroxide solution with the mass concentration of 6%, adding trypsin according to the enzyme substrate concentration ratio of 0.7% at the temperature of 38 ℃, and carrying out enzymolysis for 40min to obtain a secondary enzymolysis liquid;
(4) centrifuging the secondary enzymolysis liquid, and taking supernatant;
(5) and (3) adding an embedding wall material (sodium carboxymethylcellulose and beta-cyclodextrin in a mass ratio of 1: 3) into the supernatant obtained in the step (4) in a mass volume ratio of 10%, grinding and homogenizing for 2h, then carrying out vacuum concentration at 50 ℃, and then carrying out spray drying to obtain the microencapsulated royal jelly enzymatic hydrolysis polypeptide according to the application.
Example 4
The microencapsulated royal jelly enzymatic polypeptide is prepared by adopting the following preparation method of the microencapsulated royal jelly enzymatic polypeptide according to the application:
(1) mixing royal jelly and water in a weight ratio of 1:0.9 under ultrasonic condition, and centrifuging to obtain supernatant;
(2) adding pepsin into the supernatant obtained in the step (1) according to the concentration ratio of 0.3% of enzyme substrate, and carrying out enzymolysis for 50min at the temperature of 42 ℃ to obtain primary enzymolysis liquid;
(3) adjusting the pH of the primary enzymolysis liquid to 8 by adopting a sodium hydroxide solution with the mass concentration of 7%, adding trypsin according to the concentration ratio of 0.5% of enzyme substrate at the temperature of 45 ℃, and carrying out enzymolysis for 50min to obtain a secondary enzymolysis liquid;
(4) centrifuging the secondary enzymolysis liquid, and taking supernatant;
(5) and (3) adding an embedding wall material (sodium carboxymethylcellulose and beta-cyclodextrin in a mass ratio of 1: 2) into the supernatant obtained in the step (4) in an 8% mass-volume ratio, grinding and homogenizing for 6h, then carrying out vacuum concentration at 40 ℃, and then carrying out spray drying to obtain the microencapsulated royal jelly enzymatic polypeptide according to the application.
Example 5
The following method of preparing solid powder comprising microencapsulated royal jelly enzymatic polypeptide according to the present application was used to prepare solid powder according to the present application:
100 parts by weight of the microencapsulated royal jelly enzymatic polypeptide obtained in example 1, 80 parts by weight of the melittin, 25 parts by weight of maltodextrin, and 10 parts by weight of isomalt were uniformly mixed, thereby obtaining the solid powder containing the microencapsulated royal jelly enzymatic polypeptide.
Wherein, the bee larva peptide is prepared by the following method: crushing queen bee embryos, sieving by a 30-mesh sieve, mixing with water according to the mass ratio of 1:10 to prepare slurry, adding alkaline protease with the concentration ratio of 2% to a substrate, and carrying out enzymolysis for 4 hours at the temperature of 45 ℃ to obtain the queen bee embryo peptide.
Example 6
The following method of preparing solid powder comprising microencapsulated royal jelly enzymatic polypeptide according to the present application was used to prepare solid powder according to the present application:
120 parts by weight of the microencapsulated royal jelly enzymatic polypeptide obtained in example 2, 70 parts by weight of the melittin, 35 parts by weight of maltodextrin, and 15 parts by weight of isomalt were uniformly mixed, thereby obtaining the solid powder containing the microencapsulated royal jelly enzymatic polypeptide.
Wherein, the bee larva peptide is prepared by the following method: crushing queen bee embryos, sieving by a 30-mesh sieve, mixing with water according to the mass ratio of 1:12 to prepare slurry, adding alkaline protease with the substrate concentration ratio of 3%, and performing enzymolysis for 6 hours at the temperature of 40 ℃ to obtain the queen bee embryo peptide.
Example 7
The following method of preparing solid powder comprising microencapsulated royal jelly enzymatic polypeptide according to the present application was used to prepare solid powder according to the present application:
110 parts by weight of the microencapsulated royal jelly enzymatic polypeptide obtained in example 3, 60 parts by weight of the melittin, 20 parts by weight of maltodextrin, and 15 parts by weight of isomalt were uniformly mixed, thereby obtaining the solid powder containing the microencapsulated royal jelly enzymatic polypeptide.
Wherein, the bee larva peptide is prepared by the following method: crushing queen bee embryos, sieving by a 30-mesh sieve, mixing with water according to the mass ratio of 1:8 to prepare slurry, adding alkaline protease with the concentration ratio of 2% to a substrate, and carrying out enzymolysis for 2 hours at the temperature of 50 ℃ to obtain the queen bee embryo peptide.
Example 8
The following method of preparing solid powder comprising microencapsulated royal jelly enzymatic polypeptide according to the present application was used to prepare solid powder according to the present application:
the solid powder containing the microencapsulated royal jelly enzymatic polypeptide was obtained by uniformly mixing 110 parts by weight of the microencapsulated royal jelly enzymatic polypeptide obtained in example 4, 80 parts by weight of the melittin, 30 parts by weight of the maltodextrin, and 13 parts by weight of the isomalt.
Wherein, the bee larva peptide is prepared by the following method: crushing queen bee embryos, sieving by a 30-mesh sieve, mixing with water according to the mass ratio of 1:10 to prepare slurry, adding alkaline protease with the concentration ratio of 1% to a substrate, and carrying out enzymolysis for 5 hours at the temperature of 45 ℃ to obtain the queen bee embryo peptide.
Example 9
A solid powder according to the present application was prepared in the same preparation method as example 5, except that 0.3 parts by weight of D-mannitol and 0.15 parts by weight of nobiletin were further mixed in the solid powder.
Example 10
A solid powder according to the present application was prepared in the same preparation method as example 5, except that 0.2 parts by weight of D-mannitol and 0.1 parts by weight of nobiletin were further mixed in the solid powder.
Example 11
A solid powder according to the present application was prepared in the same preparation method as example 5, except that 0.4 parts by weight of D-mannitol and 0.2 parts by weight of nobiletin were further mixed in the solid powder.
Comparative example 1
Except that the enzymatic hydrolysis of step (2) was not performed, the microencapsulated royal jelly enzymatic hydrolyzed polypeptide was obtained in the same manner as in example 1. Also, a solid powder was further prepared in the same manner as in example 5.
Comparative example 2
Except that the enzymatic hydrolysis of step (3) was not performed, the microencapsulated royal jelly enzymatic hydrolyzed polypeptide was obtained in the same manner as in example 1. Also, a solid powder was further prepared in the same manner as in example 5.
Comparative example 3
With the exception of not adding an embedding wall material in the step (5), a royal jelly enzymatic hydrolysis polypeptide powder was prepared in the same manner as in example 1. And a solid powder was obtained in the same manner as in example 5 using this powder.
Comparative example 4
The solid powder was prepared in the same manner as in example 5, except that the microencapsulated royal jelly enzymatic polypeptide was not contained in the solid powder.
Comparative example 5
The solid powder was prepared in the same manner as in example 5, except that the peptide of Apis cerana was not contained in the solid powder.
Comparative example 6
The solid powder was prepared in the same manner as in example 5, except that 0.1 parts by weight of D-mannitol and 0.05 parts by weight of nobiletin were further mixed in the solid powder.
Comparative example 7
The solid powder was prepared in the same manner as in example 5, except that 0.6 parts by weight of D-mannitol and 0.3 parts by weight of nobiletin were further mixed in the solid powder.
Experimental examples
The beneficial effects of the present application are further illustrated by experimental data as follows:
evaluation of safety
The solid powders according to the present application prepared in examples 5 to 11 and the solid powders prepared in comparative examples 1 to 7 were tested for acute toxicity using SPF-grade Kunming mice.
Oral acute toxicity test (MTD) in mice: 280 SPF-grade Kunming mice of 18 g-22 g are selected and averagely divided into 14 groups, each group comprises 20 mice, the number of the mice is half that of the mice, and the animals are fasted for 16 hours before the test without drinking water. 50.0g of each of the samples of examples 5 to 11 and comparative examples 1 to 7 was weighed, added with pure water to 100mL, stirred uniformly to prepare a suspension with a concentration of 500mg/mL, and then the mice were gavaged for 2 times (6 h intervals) within 24h, wherein the gavage amount is 0.4mL/20g BW each time, and the total dose is 20g/kg BW. And observing and recording the poisoning expression of the animals after the gavage. Weigh once a week, observe the two week period (14 days), dissect animals for gross observation at the end of the trial. The test substance is evaluated for acute toxicity according to toxicity grading standards. The results are shown in table 1 below.
[ Table 1] results of acute toxicity test in mice
| Death situation (only) | |
| Example 5 | 0 |
| Example 6 | 0 |
| Example 7 | 0 |
| Example 8 | 0 |
| Example 9 | 0 |
| Example 10 | 0 |
| Example 11 | 0 |
| Comparative example 1 | 0 |
| Comparative example 2 | 0 |
| Comparative example 3 | 0 |
| Comparative example 4 | 0 |
| Comparative example 5 | 0 |
| Comparative example 6 | 0 |
| Comparative example 7 | 0 |
As can be seen from the above Table 1, no significant toxic manifestation was observed in the samples of examples and comparative examples, and no death occurred during the observation period. The acute oral toxicity (MTD) of the tested sample to mice of two sexes is more than 20g/kg & BW, and the test sample belongs to a non-toxic grade according to the acute toxicity grading of food safety toxicology evaluation program and method (2003 edition), and shows great safety.
Evaluation of efficacy
The solid powders according to the present application prepared in examples 5 to 11 and the solid powders prepared in comparative examples 1 to 7 were tested for fatigue resistance using SPF-grade Kunming mice.
The selected SPF-grade Kunming male mice 420 with the weight of 18 g-22 g are averagely divided into 14 groups, each group comprises 30 mice, and the number of the mice is half of that of the mice. Samples of examples 5 to 11 and comparative examples 1 to 7 were each prepared into a 50% strength solution with pure water, and 14 groups of mice were each subjected to intragastric gavage once a day for 30 consecutive days in a volume of 0.2mL/10 g.BW.
Each group of test mice was randomly divided into three subgroups of 10 mice each, and the weight swimming test, serum urea measurement, and liver glycogen measurement were performed according to the following methods, and the results are shown in Table 2 below.
1. Weight bearing swimming test
After 30 minutes of the last test sample given to the mouse, the mouse is placed in a swimming box, and the tail root of the mouse bears a 5% weight lead skin, the water depth is about 30cm, and the water temperature is 25 +/-1.0 ℃. The time from the start of swimming to death of the mouse was recorded as weight-bearing swimming time (seconds).
2. Serum urea assay
After 30 minutes of last test sample feeding, the mice swim in water with the temperature of 30 ℃ for 90 minutes without load, after 60 minutes of rest, the eyeballs are pulled out for blood sampling, after standing for 3 hours at 4 ℃, the blood is centrifuged at 2000 rpm for 15 minutes, and serum urea (mmol/L) is measured by a full-automatic biochemical analyzer for serum.
3. Liver glycogen assay (anthrone method)
After 30 minutes of the last time the mouse was tested for the sample, the animal was sacrificed, the liver was rinsed with physiological saline and then blotted dry with filter paper, 100mg of liver was weighed, and liver glycogen measurement (mg/g liver tissue) was performed using a liver glycogen measurement kit.
[ Table 2] results of efficacy measurement of each test sample
Table 2 the results show that:
1. compared with each comparative example, the mouse weight-bearing swimming time of the samples of the gavage examples 5 to 11 is obviously prolonged (P <0.01), the serum urea is obviously reduced (P <0.01), and the liver glycogen storage amount is obviously improved (P <0.05), which shows that the solid powder containing the microencapsulated royal jelly enzymolysis polypeptide prepared by each example of the application has good functions of relieving fatigue and enhancing physical strength.
2. Compared with comparative examples 1 to 3, the example 5 has longer swimming time, lower serum urea and higher liver glycogen storage, which shows that the two-stage enzymolysis for preparing the royal jelly enzymolysis polypeptide and the microencapsulation can improve the nutritional efficacy of the royal jelly.
3. Compared with comparative examples 4 and 5, the mouse weight-bearing swimming time of the sample of the gavage example 5 is obviously prolonged (P <0.05), the serum urea is obviously reduced (P <0.05), and the liver glycogen storage amount is obviously improved (P <0.05), which indicates that when the microencapsulated royal jelly enzymatic polypeptide and the bee embryo peptide in the solid powder containing the microencapsulated royal jelly enzymatic polypeptide prepared by the embodiments of the application are synergistically used, the solid powder has an effect superior to single use, and the function of relieving physical fatigue is more obvious.
4. Examples 9 to 11 further added specific amounts of D-mannitol and nobiletin, which further extended the weight-bearing swimming time of mice, reduced serum urea and increased liver glycogen stores relative to example 5. However, referring to comparative examples 6 and 7, when D-mannitol and nobiletin outside the contents defined in the present application were used, the efficacy of example 5 was not improved but rather decreased.
The above-described embodiments of the present application are only examples of the present application and should not be construed as limiting the present application, and those skilled in the art can make modifications without inventive contribution as required after reading the present specification, however, any modifications, equivalents, improvements, etc. within the spirit and principle of the present application should be included in the scope of the present application.
Claims (10)
1. A preparation method of microencapsulated royal jelly enzymatic hydrolysis polypeptide is characterized by comprising the following steps:
(1) mixing royal jelly with water, and centrifuging to obtain supernatant;
(2) adding pepsin into the supernatant obtained in the step (1) for enzymolysis to obtain primary enzymolysis liquid;
(3) adding trypsin into the primary enzymolysis liquid for enzymolysis to obtain secondary enzymolysis liquid;
(4) centrifuging the secondary enzymolysis liquid, and taking supernatant; and
(5) and (4) adding an embedding wall material into the supernatant obtained in the step (4), and grinding and homogenizing to obtain the microencapsulated royal jelly enzymatic hydrolysis polypeptide.
2. The preparation method according to claim 1, wherein in the step (1), the weight ratio of royal jelly to water is 1 (0.8-1);
the mixing in step (1) is carried out under ultrasonic conditions.
3. The preparation method according to claim 1, wherein the enzymolysis in the step (2) is: carrying out enzymolysis for 40-60 min at the temperature of 38-45 ℃;
in the step (2), the concentration ratio of the enzyme substrate can be 0.3-0.5%;
the enzymolysis in the step (3) is as follows: adjusting the pH value of the system to 8, adding trypsin at the temperature of 38-45 ℃, and carrying out enzymolysis for 40-60 min;
in the step (3), sodium hydroxide solution with the mass concentration of 5-8% is adopted to adjust the pH value; and
in the step (3), the concentration ratio of the enzyme substrate may be 0.5% to 0.7%.
4. The preparation method according to claim 1, wherein in the step (5), the addition amount of the embedding wall material is 0.5 to 10% by mass/volume;
the embedding wall material comprises sodium carboxymethylcellulose and beta-cyclodextrin in a mass ratio of 1 (0.1-3);
the grinding homogenization time is 2-8 h.
5. The method according to claim 1, wherein the step (5) further comprises concentrating the resultant under vacuum at 40 to 50 ℃ after completion of the grinding homogenization, followed by spray drying.
6. A microencapsulated royal jelly enzymatic polypeptide obtained by the production method according to any one of claims 1 to 5.
7. A solid powder, comprising:
the microencapsulated royal jelly enzymatic polypeptide obtained by the preparation method according to any one of claims 1 to 5 or the microencapsulated royal jelly enzymatic polypeptide according to claim 6
8. The solid powder of claim 7, wherein the melittin is prepared by the following method:
pulverizing queen bee embryo, mixing with water to obtain slurry, adding alkaline protease for enzymolysis to obtain foetus Apis peptide,
preferably, the queen bee larva is crushed and sieved by a 30-mesh sieve;
preferably, queen bee larva powder is mixed with water according to the mass ratio of 1 (8-12);
preferably, the concentration ratio of the alkaline protease to the substrate can be 1-3%; and
preferably, the enzymolysis is carried out for 2 to 6 hours at the temperature of 40 to 50 ℃.
9. The solid powder of claim 7, further comprising:
0.2 to 0.4 part by weight of D-mannitol, and
0.1 to 0.2 part by weight of nobiletin,
preferably, the mass ratio of the D-mannitol to the nobiletin is 2: 1.
10. A method of preparing a solid powder according to any one of claims 7 to 9, comprising:
and uniformly mixing the components to obtain the solid powder containing the microencapsulated royal jelly enzymatic hydrolysis polypeptide.
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