CN113249422A - Preparation method of royal jelly polypeptide - Google Patents
Preparation method of royal jelly polypeptide Download PDFInfo
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- CN113249422A CN113249422A CN202110503006.0A CN202110503006A CN113249422A CN 113249422 A CN113249422 A CN 113249422A CN 202110503006 A CN202110503006 A CN 202110503006A CN 113249422 A CN113249422 A CN 113249422A
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- royal jelly
- mixed solution
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Abstract
The invention discloses a preparation method of royal jelly polypeptide, belonging to the technical field of bee product processing. The invention not only eliminates the sensitization of the royal jelly, but also is beneficial to the release of flavor amino acid and improves the taste, the hydrolysate contains a large amount of polypeptide with the molecular weight of 1kDa, most of which consists of 2-4 amino acids, such as Ile-Leu, Val-Thr and Tyr-Leu, in addition, the content of 24-methylene cholesterol in the royal jelly can be improved through enzymolysis, and the substance has various functions of reducing blood pressure, reducing cholesterol, resisting arrhythmia, relieving vascular spasm and the like, and has better effect on the auxiliary treatment of prostatic hyperplasia.
Description
Technical Field
The invention belongs to the technical field of bee product processing, and relates to a preparation method of royal jelly polypeptide.
Background
The royal jelly is thick paste secreted by the glands of the pharyngeal gland, the upper jaw gland, the back brain gland and the like of young worker bees (feeding worker bees) of 5-15 days old in a bee colony, contains rich functional protein (accounting for about 50 percent of dry matter of the royal jelly), functional lipid, acetylcholine, antibacterial peptide and other various nutritional and active ingredients, has the effects of improving immunity, resisting fatigue, resisting oxidation, resisting tumors, resisting bacteria and the like on a human body, and is a high-quality functional food raw material and a special medical food raw material. However, royal jelly is sour and astringent in taste and poor in palatability, and some people give up eating the royal jelly due to the taste. Some people take royal jelly and have mild anaphylactic reaction, and the possible reason is that the people are allergic to the main protein of the royal jelly. Therefore, the development of new royal jelly products is urgently needed, the nutritive value of the royal jelly is improved, the mouthfeel is improved, the sensitization is reduced, and the needs of people for consuming the royal jelly products are met.
MRJP1 and MRJP2 are the major allergens in royal jelly. The royal jelly is subjected to enzymolysis by protease, so that the content of allergic protein can be obviously reduced, bioactive peptide is generated, the risk of allergy caused by eating the royal jelly is effectively reduced, and the nutrition and health care functions of the royal jelly are greatly improved. Research shows that the immunoregulation function of the hydrolyzed royal jelly product is obviously improved compared with that of royal jelly. In addition, the content of active ingredients in the royal jelly can be improved through enzymolysis, so that the royal jelly has various physiological functions. Therefore, the invention hydrolyzes the main protein of royal jelly through protease to obtain micromolecule active peptide, improve the content of 24-methylene cholesterol, reduce the content of allergic protein and enhance the immunoregulation function.
Disclosure of Invention
The present invention aims to solve the above problems and provide a royal jelly polypeptide and a preparation method thereof, which can increase the content of active ingredients in royal jelly, make the royal jelly have various physiological functions, reduce the content of allergic proteins, and enhance the immune regulation function of the royal jelly.
The invention realizes the aim through the following technical scheme, and provides a preparation method of royal jelly polypeptide, which comprises the following raw material components: royal jelly, deionized water, edible citric acid, edible pepsin, edible calcium hydroxide and edible papain; the preparation method of the royal jelly polypeptide comprises the following steps:
step 1: adding deionized water into royal jelly, and fully stirring and uniformly mixing to obtain a mixed solution A;
step 2: adjusting the pH value of the mixed solution A obtained in the step 1 to 1.5 by using edible citric acid;
and step 3: adding 5% of edible pepsin into the mixed solution A in the step 2 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution B;
and 4, step 4: placing the mixed solution B obtained in the step 3 at a constant temperature of 37 ℃ for enzymolysis to obtain hydrolysate C;
and 5: adjusting the pH value of the hydrolysate C in the step 4 to 7.0 by using edible calcium hydroxide to obtain a mixed solution D;
step 6: adding 5% of edible papain into the mixed solution D obtained in the step 5 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution E;
and 7: placing the mixed solution E obtained in the step 6 at a constant temperature of 55 ℃, and carrying out enzymolysis to obtain royal jelly enzymatic hydrolysate;
and 8: after the enzymolysis is finished, the royal jelly enzymolysis liquid is placed at 85 ℃ to inactivate the enzyme.
Preferably, the royal jelly is mixed with the deionized water according to the mass ratio of 1:8-12 in the step 1.
Preferably, the mixed solution B in the step 4 is placed at a constant temperature for enzymolysis for 7-8 h.
Preferably, the constant-temperature enzymolysis time of the mixed solution E in the step 7 is 2-4 h.
Preferably, the royal jelly enzymatic hydrolysate in step 8 is kept at 85 ℃ for 13-17 min.
The invention has the beneficial effects that:
the invention not only eliminates the sensitization of royal jelly, but also is beneficial to the release of flavor amino acid and improves the taste. The hydrolysate contains a large amount of polypeptides with the molecular weight of less than 1kDa, and most of the polypeptides consist of 2-4 amino acids, such as Ile-Leu, Val-Thr and Tyr-Leu, and the peptides of the type have proved to have strong capability of resisting grease oxidation; in addition, the comparison of database results shows that about 70 polypeptides are not identified by the predecessors, the content of 24-methylene cholesterol in royal jelly can be increased through enzymolysis, and the substance has multiple functions of reducing blood pressure, reducing cholesterol, resisting arrhythmia, relieving vascular spasm and the like, and has a good effect on adjuvant therapy of prostatic hyperplasia.
Drawings
FIG. 1 shows the behavior of the solution before enzymolysis;
FIG. 2 shows the behavior of the solution after enzymolysis;
FIG. 3 is a blank map of the hydrolysis of protease itself;
FIG. 4 is a blank comparison chart of the self-hydrolysis of royal jelly;
FIG. 5 is a diagram showing the results of royal jelly enzymolysis;
FIG. 6 is a mass spectrum fragment diagram of valine-threonine of royal jelly enzymatic hydrolysate;
FIG. 7 is a fragment diagram of tyrosine-leucine mass spectrum of royal jelly enzymatic hydrolysate;
FIG. 8 is a mass spectrum fragment diagram of isoleucine-leucine of royal jelly enzymatic hydrolysate;
FIG. 9 is a mass spectrum fragment diagram of glutamine-isoleucine-valine as an enzymolysis product of royal jelly;
FIG. 10 is a fragment diagram of mass spectrum of glycine-phenylalanine as an enzymatic hydrolysate of royal jelly;
FIG. 11 is a mass spectrum fragment diagram of proline-leucine as an enzymolysis product of royal jelly;
FIG. 12 is a gas chromatography mass spectrum TIC chart of determination of 24-methylene cholesterol in royal jelly;
FIG. 13 is a mass spectrum of 24-methylene cholesterol in royal jelly.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A royal jelly polypeptide comprises the following raw material components:
royal jelly, deionized water, edible citric acid, edible pepsin, edible calcium hydroxide and edible papain.
A preparation method of royal jelly polypeptide comprises the following steps:
step 1: adding deionized water into royal jelly, mixing the royal jelly and the deionized water according to the mass ratio of 1:8, and fully stirring and uniformly mixing to obtain a mixed solution A;
step 2: adjusting the pH value of the mixed solution A obtained in the step 1 to 1.5 by using edible citric acid;
and step 3: adding 5% of edible pepsin into the mixed solution A in the step 2 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution B;
and 4, step 4: carrying out enzymolysis on the mixed solution B obtained in the step 3 at a constant temperature of 37 ℃ for 7 hours to obtain hydrolysate C;
and 5: adjusting the pH value of the hydrolysate C in the step 4 to 7.0 by using edible calcium hydroxide to obtain a mixed solution D;
step 6: adding 5% of edible papain into the mixed solution D obtained in the step 5 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution E;
and 7: placing the mixed solution E obtained in the step 6 at a constant temperature of 55 ℃ for enzymolysis for 2 hours to obtain royal jelly enzymolysis solution;
and 8: after the enzymolysis is finished, the royal jelly enzymolysis liquid is kept at 85 ℃ for 13min to inactivate the enzyme.
Example 2
A royal jelly polypeptide comprises the following raw material components:
royal jelly, deionized water, edible citric acid, edible pepsin, edible calcium hydroxide and edible papain.
A preparation method of royal jelly polypeptide comprises the following steps:
step 1: adding deionized water into royal jelly, mixing the royal jelly and the deionized water according to the mass ratio of 1:10, and fully stirring and uniformly mixing to obtain a mixed solution A;
step 2: adjusting the pH value of the mixed solution A obtained in the step 1 to 1.5 by using edible citric acid;
and step 3: adding 5% of edible pepsin into the mixed solution A in the step 2 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution B;
and 4, step 4: carrying out enzymolysis on the mixed solution B obtained in the step 3 at a constant temperature of 37 ℃ for 7 hours to obtain hydrolysate C;
and 5: adjusting the pH value of the hydrolysate C in the step 4 to 7.0 by using edible calcium hydroxide to obtain a mixed solution D;
step 6: adding 5% of edible papain into the mixed solution D obtained in the step 5 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution E;
and 7: placing the mixed solution E obtained in the step 6 at a constant temperature of 55 ℃ for enzymolysis for 3 hours to obtain royal jelly enzymolysis solution;
and 8: after the enzymolysis is finished, the royal jelly enzymolysis liquid is placed at 85 ℃ and kept for 15min to inactivate the enzyme.
Example 3
A royal jelly polypeptide comprises the following raw material components:
royal jelly, deionized water, edible citric acid, edible pepsin, edible calcium hydroxide and edible papain.
A preparation method of royal jelly polypeptide comprises the following steps:
step 1: adding deionized water into royal jelly, mixing the royal jelly and the deionized water according to the mass ratio of 1:12, and fully stirring and uniformly mixing to obtain a mixed solution A;
step 2: adjusting the pH value of the mixed solution A obtained in the step 1 to 1.5 by using edible citric acid;
and step 3: adding 5% of edible pepsin into the mixed solution A in the step 2 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution B;
and 4, step 4: carrying out enzymolysis on the mixed solution B obtained in the step 3 at a constant temperature of 37 ℃ for 8 hours to obtain hydrolysate C;
and 5: adjusting the pH value of the hydrolysate C in the step 4 to 7.0 by using edible calcium hydroxide to obtain a mixed solution D;
step 6: adding 5% of edible papain into the mixed solution D obtained in the step 5 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution E;
and 7: placing the mixed solution E obtained in the step 6 at the constant temperature of 55 ℃ for enzymolysis for 4 hours to obtain royal jelly enzymolysis solution;
and 8: after the enzymolysis is finished, the royal jelly enzymolysis liquid is placed at 85 ℃ and kept for 17min to inactivate the enzyme.
(1) Referring to fig. 1 and 2:
the flocculent precipitate of protein after enzymolysis is gradually reduced, the sample is gradually bright and clear, and part of the substances which are not dissolved in water, such as 10-HDA, are precipitated to the bottom, which proves that under the condition, the main protein in the royal jelly is subjected to enzymolysis to form the short peptide which is easy to dissolve, and the functional lipid substance which is combined with the protein is released, and is gradually precipitated to the bottom of the fermentation bottle because the substance is not dissolved in water.
(2) By analyzing the main characteristic substance 10-HDA of the royal jelly by applying the GB9697-2008 method, the content of the royal jelly is 1.85 percent before enzymolysis, 1.83 percent after enzymolysis and basically unchanged. The enzymolysis process has no influence on the micromolecular nutritional characteristic components of the royal jelly.
(3) Referring to fig. 3, 4, 5:
analyzing the royal jelly after enzymolysis by high performance liquid chromatography/high resolution mass spectrometry (UPLC-Q/TOFMS method). Comparing fig. 3, 4 and 5, it can be seen that the main enzymatic hydrolysis products marked in the figures are not self-digestion of the protease and royal jelly, but are royal jelly peptides generated after the enzymatic hydrolysis of royal jelly. As can be seen from fig. 6, 7, 8, 9, 10 and 11, the molecular weight of the enzymatic product is mostly below 1kDa, mostly consisting of 2-4 amino acids, and a small amount of more than 5 amino acids, and fig. 6, 7, 8, 9, 10 and 11 show the partially identified polypeptide structures.
(4) Referring to fig. 12, 13:
it can be seen that the content of small molecular substance 24-methylene cholesterol in royal jelly before enzymolysis is 0.188mg/g, and the content of the substance after enzymolysis is 0.263mg/g, which is increased by about 40% (dry matter proportion).
24-methylene cholesterol mainly forms a compound with royal jelly main protein in royal jelly, and can effectively promote the release of the substance from the protein through enzymolysis. The research shows that the substance can reduce the concentration of low-density lipoprotein cholesterol in plasma, has multiple functions of reducing blood pressure, resisting arrhythmia, relieving vascular spasm and the like, and has better effect on the adjuvant therapy of the prostatic hyperplasia.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (5)
1. The preparation method of the royal jelly polypeptide is characterized in that the royal jelly polypeptide comprises the following raw material components:
royal jelly, deionized water, edible citric acid, edible pepsin, edible calcium hydroxide and edible papain;
the preparation method of the royal jelly polypeptide comprises the following steps:
step 1: adding deionized water into royal jelly, and fully stirring and uniformly mixing to obtain a mixed solution A;
step 2: adjusting the pH value of the mixed solution A obtained in the step 1 to 1.5 by using edible citric acid;
and step 3: adding 5% of edible pepsin into the mixed solution A in the step 2 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution B;
and 4, step 4: placing the mixed solution B obtained in the step 3 at a constant temperature of 37 ℃ for enzymolysis to obtain hydrolysate C;
and 5: adjusting the pH value of the hydrolysate C in the step 4 to 7.0 by using edible calcium hydroxide to obtain a mixed solution D;
step 6: adding 5% of edible papain into the mixed solution D obtained in the step 5 according to the mass of the royal jelly, and fully and uniformly mixing to obtain mixed solution E;
and 7: placing the mixed solution E obtained in the step 6 at a constant temperature of 55 ℃, and carrying out enzymolysis to obtain royal jelly enzymatic hydrolysate;
and 8: after the enzymolysis is finished, the royal jelly enzymolysis liquid is placed at 85 ℃ to inactivate the enzyme.
2. The method for preparing a royal jelly polypeptide according to claim 1, which comprises: in the step 1, the royal jelly and the deionized water are mixed according to the mass ratio of 1: 8-12.
3. The method for preparing a royal jelly polypeptide according to claim 1, which comprises: and 4, placing the mixed liquor B in the step 4 for enzymolysis at constant temperature for 7-8 h.
4. The method for preparing a royal jelly polypeptide according to claim 1, which comprises: and (7) carrying out constant-temperature enzymolysis on the mixed solution E for 2-4 h.
5. The method for preparing a royal jelly polypeptide according to claim 1, which comprises: and 8, keeping the bee royal jelly enzymatic hydrolysate at 85 ℃ for 13-17 min.
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| CN202110503006.0A CN113249422A (en) | 2021-05-10 | 2021-05-10 | Preparation method of royal jelly polypeptide |
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| CN202110503006.0A CN113249422A (en) | 2021-05-10 | 2021-05-10 | Preparation method of royal jelly polypeptide |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113647591A (en) * | 2021-08-20 | 2021-11-16 | 山东丰采健康产业有限公司 | Royal jelly zymolyte and preparation method and application thereof |
| CN117064996A (en) * | 2023-10-08 | 2023-11-17 | 江苏鸿祺生物科技有限公司 | Composition for protecting prostate and preparation method thereof |
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| JP2008056645A (en) * | 2006-09-04 | 2008-03-13 | Yumedeika Kk | Anti-oxidant peptide obtained by reaction of protein in enzymatically treated royal jelly with polypeptide and method for producing the same |
| JP2008194031A (en) * | 2007-01-17 | 2008-08-28 | Yamada Bee Farm Corp | Stabilization of enzyme-treated royal jelly |
| CN108998488A (en) * | 2018-07-31 | 2018-12-14 | 巢氏健康生物科技股份有限公司 | A kind of activity royal jelly OCO polypeptide freeze-dried powder and preparation method thereof |
| CN112662721A (en) * | 2020-12-25 | 2021-04-16 | 夏晓勤 | Protein polypeptide for resisting aging and preparation method thereof |
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2021
- 2021-05-10 CN CN202110503006.0A patent/CN113249422A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008056645A (en) * | 2006-09-04 | 2008-03-13 | Yumedeika Kk | Anti-oxidant peptide obtained by reaction of protein in enzymatically treated royal jelly with polypeptide and method for producing the same |
| JP2008194031A (en) * | 2007-01-17 | 2008-08-28 | Yamada Bee Farm Corp | Stabilization of enzyme-treated royal jelly |
| CN108998488A (en) * | 2018-07-31 | 2018-12-14 | 巢氏健康生物科技股份有限公司 | A kind of activity royal jelly OCO polypeptide freeze-dried powder and preparation method thereof |
| CN112662721A (en) * | 2020-12-25 | 2021-04-16 | 夏晓勤 | Protein polypeptide for resisting aging and preparation method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113647591A (en) * | 2021-08-20 | 2021-11-16 | 山东丰采健康产业有限公司 | Royal jelly zymolyte and preparation method and application thereof |
| CN113647591B (en) * | 2021-08-20 | 2023-11-10 | 山东丰采健康产业有限公司 | Royal jelly zymolyte and preparation method and application thereof |
| CN117064996A (en) * | 2023-10-08 | 2023-11-17 | 江苏鸿祺生物科技有限公司 | Composition for protecting prostate and preparation method thereof |
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