CN114891851A - Biological selenium active peptide and preparation method and application thereof - Google Patents
Biological selenium active peptide and preparation method and application thereof Download PDFInfo
- Publication number
- CN114891851A CN114891851A CN202210813744.XA CN202210813744A CN114891851A CN 114891851 A CN114891851 A CN 114891851A CN 202210813744 A CN202210813744 A CN 202210813744A CN 114891851 A CN114891851 A CN 114891851A
- Authority
- CN
- China
- Prior art keywords
- selenium
- peptide
- active peptide
- biological
- enzymolysis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/10—Process efficiency
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- General Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Water Supply & Treatment (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention relates to the technical field of deep processing of plant protein and trace elements, in particular to biological selenium active peptide and a preparation method and application thereof, wherein the biological selenium active peptide is prepared by germinating soaked selenium-rich seeds; then, by selecting the combination of amylase, alkaline protease and aminopeptidase, controlling the adding sequence of the enzymes and the adding interval time, the enzymolysis efficiency can be greatly improved, selenoprotein in the germinated pulp is subjected to enzymolysis to form the biological selenium active peptide of small peptide molecules, so that the peptide content of the finally prepared biological selenium active peptide with the molecular weight of below 1000 daltons reaches more than 98wt%, the obtained biological selenium active peptide is dissolved in water, the small peptide molecules can quickly convey selenium element to cells to participate in life movement, and the absorption rate is greatly improved.
Description
Technical Field
The invention relates to the technical field of deep processing of plant protein and trace elements, in particular to a biological selenium active peptide and a preparation method and application thereof.
Background
Selenium is an important component of some antioxidant enzymes and selenoprotein P in animals and human bodies, plays a role in balancing oxidation-reduction atmosphere in the bodies, and researches prove that the selenium has the function of improving the immunity of the bodies and is a research hotspot for international immunity influence and cancer prevention, so the selenium can be used as a trace additive of animal feed, and trace element fertilizer is added into plant fertilizer to improve the selenium content of agricultural and sideline products. Selenium has been used as an essential trace element in humans. The scientist determines: some diseases, especially tumors, hypertension, endocrine and metabolic diseases, diabetes and senile constipation are all related to selenium deficiency. Selenium is honored by the medical and nutritional communities at home and abroad as the "fire of life", enjoying the reputations of "longevity element", "anticancer king", "heart guard spirit", "natural antidote" and the like. Selenium is one ten million in human tissues, but the selenium determines the existence of life, and the selenium has great effect on human health and cannot be replaced by other substances. The selenium deficiency can directly lead to the reduction of the immunity of human bodies, and more than forty diseases threatening the health and the life of human bodies are proved to be related to the selenium deficiency of human bodies by clinical medicine, such as cancers, cardiovascular diseases, liver diseases, cataract, pancreatic diseases, diabetes, reproductive system diseases and the like. According to the examination of experts, people need to supplement selenium for a lifetime.
The total amount of selenium in adult is 6-20 mg. Selenium is distributed throughout tissues, organs and body fluids, with the highest concentration in the kidney. Mainly exists in the form of selenium and protein combined complex in the tissue. Selenium is mainly absorbed in small intestine, and the absorption rate of selenium in food by human body is 60% -80%. The selenium absorbed by intestinal tract is metabolized and then the majority is discharged via urine. Urine selenium is a good index for judging the excess and deficiency of selenium in human bodies. Other ways of selenium excretion are feces and sweat. The absorption, transport, excretion, storage and distribution of selenium in the body are influenced by many external factors, as are the sex, age, health and the presence of compounds such as sulphur, heavy metals, vitamins in the food. Animal experiments show that selenium is mainly absorbed in the duodenum, the jejunum and ileum are slightly absorbed, and the stomach is not absorbed. The selenium excretion via urine accounts for 50-60% of the selenium excretion, and when high dietary selenium is ingested, the selenium excretion increases, otherwise, the excretion decreases, and the kidney plays a regulating role.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide a biological selenium active peptide which is easy to be absorbed and utilized by human body and has no side effect, and a preparation method and application thereof.
A preparation method of biological selenium active peptide comprises the following steps:
(1) soaking the selenium-rich plant seeds in water, taking out the selenium-rich plant seeds, spraying water for 2-3 times per day for germination, discharging after 6-7 days of germination to obtain seed germination raw materials, and then stirring to obtain a slurry;
(2) and enzymolysis: sequentially adding amylase accounting for 0.5-0.8% of the weight of the seed germination raw material, alkaline protease accounting for 1-1.5% of the weight of the seed germination raw material and aminopeptidase accounting for 0.5-0.8% of the weight of the seed germination raw material into the slurry, wherein the adding time interval of two adjacent enzymes is 0.4-0.6h, stirring for enzymolysis, and after the last enzyme is added, continuously stirring for enzymolysis for 3-3.5 h; the temperature of stirring and enzymolysis is kept between 55 and 57 ℃;
(3) enzyme deactivation;
(4) carrying out solid-liquid separation;
(5) filtering the separated liquid obtained in the step (4) by using a nanofiltration membrane of 0.8-1.2nm, reserving the permeate, then filtering by using a nanofiltration membrane of 0.2-0.3nm, and reserving the membrane retentate;
(6) adding organic selenium into the membrane trapped fluid obtained in the step (5) to carry out peptide selenium chelation;
(7) and (4) concentrating and drying the chelating solution obtained in the step (6).
Optionally, adding selenium-rich plant seeds into purified water 5-6 times of the weight of the selenium-rich plant seeds, and soaking for 12-18 h; and/or
The germination temperature is 15-20 deg.C, and the humidity is 65-70%.
Optionally, the enzyme activity of the amylase is 17-23U/mg;
the enzyme activity of the alkaline protease is 25-31U/mg;
the enzyme activity of the aminopeptidase is 22-28U/mg.
Optionally, in the step (3), the temperature of the enzymatic hydrolysate obtained in the step (2) is increased to 90-95 ℃, and enzyme deactivation is carried out for 10-15 min.
Optionally, in the step (4), in the solid-liquid separation step, a horizontal screw centrifuge is used for filtering to remove solid matters, a plate-and-frame filter is used for filtering, and finally a three-phase centrifuge is used for centrifugal purification;
the rotating speed of the horizontal screw centrifuge is 3600 revolutions per minute;
the aperture of a filter plate in the plate and frame filter is 0.3-0.4 μm;
the rotational speed of the three-phase centrifuge is 18000 rpm.
Optionally, in the step (6), the addition amount of the organic selenium is 0.06-0.07 per mill of the weight of the seed germination raw material.
Optionally, in the step (6), the peptide selenium is chelated under the following conditions: stirring at 45-50 deg.C for 30-45min at 36-45 r/min.
The biological selenium active peptide is prepared by a preparation method of the biological selenium active peptide, wherein in peptide molecules of the biological selenium active peptide, the mass percentage of peptide molecules with the relative molecular weight of less than or equal to 1000 daltons in the total peptide molecules is more than 98%, and the content of the peptide molecules in the biological selenium active peptide is more than 95 wt%.
Optionally, in the biological selenium active peptide, the total content of dipeptide and tripeptide accounts for more than 45wt% of the total peptide molecule content.
A biological selenium active peptide prepared by the preparation method of the biological selenium active peptide or the application of the biological selenium active peptide in preparing feed additives, nutritional formula foods, health products, special medical foods or medicines.
The technical scheme of the invention has the following advantages:
1. according to the preparation method of the biological selenium active peptide, the soaked selenium-rich seeds germinate to realize biotransformation, so that the yield of the biological selenium active peptide can be improved, and particularly, the contents of dipeptide and tripeptide in the product can be obviously improved; then, by selecting the combination of amylase, alkaline protease and aminopeptidase, proteolysis of protein in the slurry after seed germination and smashing can be carried out to form small peptide molecules, and meanwhile, the adding sequence and adding interval time of the enzyme are controlled, so that on one hand, the situation that some enzyme in the amylase, the alkaline protease and the aminopeptidase interferes with other enzyme to influence part of the enzyme to play a role is avoided, and the method can greatly improve the enzymolysis efficiency, so that the peptide content of the finally prepared biological selenium active peptide with the relative molecular weight of less than 1000 daltons reaches more than 98wt%, the obtained biological selenium active peptide is further dissolved in water, the small peptide molecules can quickly convey selenium elements to cells to participate in life movement, and the absorption rate and the utilization rate are greatly improved; on the other hand, the small peptide molecules can be prevented from being excessively hydrolyzed into single amino acid by partial enzyme, and the method can greatly reduce the content of the single amino acid in the small peptide molecules so as to avoid low selenium element content in the biological selenium active peptide and reduce the yield of the small peptide molecules due to the fact that the single amino acid cannot chelate organic selenium. Therefore, the small peptide molecules obtained by the method are complete protein active peptides, and compared with other chelates, the small peptide molecules have better chelation on selenium, are beneficial to chelating free selenium, and improve the utilization rate of selenium;
furthermore, a nanofiltration membrane is adopted for filtering, so that the molecular weight of the product can be controlled, and the absorption rate is improved;
in addition, the biological selenium active peptide from the plant has no side effect on the human body, and the biological selenium active peptide prepared in the preparation process has no toxic or side effect substances.
2. The alkaline protease and the aminopeptidase can reduce the molecular weight of the product, and the aminopeptidase can selectively remove hydrophobic amino acid residues and remove bitter groups at two ends of peptide bonds, so that the product tastes better, and small peptide molecules in feed liquid can be chelated with selenium more easily. The amylase can degrade starch in plant protein and is beneficial to degrading protein.
3. In the solid-liquid separation step, a horizontal screw centrifuge is firstly adopted to filter solid matters, then a plate-and-frame filter is adopted to filter the solid matters, and finally a three-phase centrifuge is adopted to carry out centrifugal purification, so that insoluble matters in enzymatic hydrolysate can be removed, and the problem of difficult filtration in the subsequent nanofiltration step is solved.
4. The biological selenium active peptide provided by the invention has the advantages that in peptide molecules of the biological selenium active peptide, the mass percentage of peptide molecules with the relative molecular weight of less than or equal to 1000 daltons in the total peptide molecules is more than 98%, the peptide molecule content in the biological selenium active peptide is more than 95wt%, the molecular structure of the biological selenium active peptide is closer to the natural form of selenium existing in organisms, the biological selenium active peptide is directly absorbed and utilized by intestinal tracts, can permeate placenta to increase selenium reserve, has high absorptivity of organisms, has no side effect, and can be widely applied to feed additives, nutritional formula foods, health products, special medical foods or medicines.
Detailed Description
The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
The alkaline protease is endonuclease; aminopeptidase is exoprotease and is obtained by fermentation of Aspergillus oryzae. Amylases, alkaline proteases, and aminopeptidases are all commercially available. The amylase, alkaline protease and aminopeptidase used in the method respectively have the enzyme activities of: 20U/mg, 28U/mg, 25U/mg; the horizontal screw centrifugal machine, the three-phase centrifugal machine and the nanofiltration membrane separation device are universal devices. The organic selenium in the following examples is a commercially available product.
Example 1
A preparation method of biological selenium active peptide comprises the following steps:
(1) adding selenium-rich plant seeds (selenium-rich wheat) into purified water which is 5 times of the weight of the selenium-rich plant seeds, soaking for 18h, taking out, spraying water for 2 times every day for germination, wherein the germination temperature is 20 ℃, the humidity is 65%, discharging after germination for 7 days to obtain a germination raw material, and then crushing to obtain a germination slurry;
(2) and enzymolysis: sequentially adding amylase accounting for 0.8 percent of the weight of the germination raw material, alkaline protease accounting for 1.5 percent of the weight of the germination raw material and aminopeptidase accounting for 0.8 percent of the weight of the germination raw material into the germination pulp, wherein the adding time interval of two adjacent enzymes is 0.6h, stirring and carrying out enzymolysis, and after the adding of the last enzyme is finished, continuously stirring and carrying out enzymolysis for 3.5 h; the temperature of stirring enzymolysis is kept at 57 ℃;
(3) and enzyme deactivation: heating the enzymolysis liquid obtained in the step (2) to 95 ℃, and keeping for 10min to inactivate enzyme;
(4) and solid-liquid separation: filtering the enzyme-inactivating liquid obtained in the step (3) by using a horizontal screw centrifuge (the rotating speed is 3600 r/min) to remove solid matters, filtering by using a plate-and-frame filter, and finally centrifuging and purifying by using a three-phase centrifuge; the aperture of a filter plate in the plate and frame filter is 0.3 mu m; the rotating speed of the three-phase centrifuge is 18000 r/min;
(5) filtering the separated liquid obtained in the step (4) by using a nanofiltration membrane of 0.8nm, reserving the permeate, then filtering by using a nanofiltration membrane of 0.2nm, and reserving the membrane retentate;
(6) adding organic selenium into the membrane trapped fluid obtained in the step (5), then heating to 45 ℃, and stirring for 45min at the stirring speed of 36 r/min; the addition amount of the organic selenium is 0.06 per mill of the weight of the added germination raw materials;
(7) and (4) concentrating the chelating solution obtained in the step (6) by using a double-effect concentrator, and spray-drying the concentrated solution to obtain the biological selenium active peptide product.
Example 2
A preparation method of biological selenium active peptide comprises the following steps:
(1) adding selenium-enriched wheat into purified water which is 5.5 times of the weight of the selenium-enriched wheat, soaking for 16h, taking out, spraying water for 2 times every day for germination, wherein the germination temperature is 17 ℃, the humidity is 67%, discharging after germination for 7 days to obtain a germination raw material, and then crushing to obtain germination slurry;
(2) and enzymolysis: sequentially adding amylase accounting for 0.6 percent of the weight of the germination raw materials, alkaline protease accounting for 1.3 percent of the weight of the germination raw materials and aminopeptidase accounting for 0.6 percent of the weight of the germination raw materials, wherein the adding time interval of two adjacent enzymes is 0.5h, stirring and carrying out enzymolysis, and after the last enzyme is added, continuously stirring and carrying out enzymolysis for 3.5 h; keeping the temperature of stirring enzymolysis at 56 ℃;
(3) and enzyme deactivation: heating the enzymolysis liquid obtained in the step (2) to 93 ℃, and keeping for 13min to inactivate enzyme;
(4) and solid-liquid separation: filtering the enzyme-inactivating liquid obtained in the step (3) by using a horizontal screw centrifuge (the rotating speed is 3600 r/min) to remove solid matters, filtering by using a plate-and-frame filter, and finally centrifuging and purifying by using a three-phase centrifuge; the aperture of a filter plate in the plate and frame filter is 0.3 mu m; the rotating speed of the three-phase centrifuge is 18000 r/min;
(5) filtering the separated liquid obtained in the step (4) by using a nanofiltration membrane of 1.0nm, reserving the permeate, then filtering by using a nanofiltration membrane of 0.2nm, and reserving the membrane retentate;
(6) adding organic selenium into the membrane trapped fluid obtained in the step (5), then heating to 48 ℃, and stirring for 37min at the stirring speed of 40 r/min; the addition amount of the organic selenium is 0.065 per mill of the weight of the added germination raw materials;
(7) and (4) concentrating the chelating solution obtained in the step (6) by using a double-effect concentrator, and spray-drying the concentrated solution to obtain the biological selenium active peptide product.
Example 3
A preparation method of biological selenium active peptide comprises the following steps:
(1) adding selenium-enriched wheat into purified water 6 times of the weight of the selenium-enriched wheat, soaking for 12h, taking out, spraying water for 3 times every day for germination, wherein the germination temperature is 15 ℃, the humidity is 70%, discharging after germination for 6 days to obtain a germination raw material, and then crushing to obtain a germination slurry;
(2) and enzymolysis: sequentially adding amylase accounting for 0.5 percent of the weight of the germination raw materials, alkaline protease accounting for 1 percent of the weight of the germination raw materials and aminopeptidase accounting for 0.5 percent of the weight of the germination raw materials into the germination slurry liquid, wherein the adding time interval of two adjacent enzymes is 0.4h, stirring and carrying out enzymolysis, and after the adding of the last enzyme is finished, continuously stirring and carrying out enzymolysis for 3 h; the temperature of stirring enzymolysis is kept at 55 ℃;
(3) and enzyme deactivation: heating the enzymolysis liquid obtained in the step (2) to 90 ℃, and keeping for 15min to inactivate enzyme;
(4) and solid-liquid separation: filtering the enzyme-inactivating liquid obtained in the step (3) by using a horizontal screw centrifuge (the rotating speed is 3600 r/min) to remove solid matters, filtering by using a plate-and-frame filter, and finally centrifuging and purifying by using a three-phase centrifuge; the aperture of a filter plate in the plate frame filter is 0.4 mu m; the rotating speed of the three-phase centrifuge is 18000 r/min;
(5) filtering the separated liquid obtained in the step (4) by using a nanofiltration membrane of 1.2nm, reserving the permeate, then filtering by using a nanofiltration membrane of 0.3nm, and reserving the membrane retentate;
(6) adding organic selenium into the membrane trapped fluid obtained in the step (5), then heating to 50 ℃, and stirring for 30min at the stirring speed of 45 r/min; the addition amount of the organic selenium is 0.07 per mill of the weight of the added germination raw materials;
(7) and (4) concentrating the chelating solution obtained in the step (6) by using a double-effect concentrator, and spray-drying the concentrated solution to obtain the biological selenium active peptide product.
Comparative example 1
The comparative example is different from example 1 in that in the enzymolysis step, amylase, alkaline protease and aminopeptidase are added simultaneously, the addition amount of the enzymes is the same as that in example 1, the total enzymolysis time is the same as that in example 1, and the enzymolysis temperature and stirring speed are the same as those in example 1.
Comparative example 2
The difference between the comparative example and the example 1 is that in the step (1), the selenium-enriched wheat is crushed and sieved by a 80-mesh sieve, then purified water which is 3 times of the weight of the selenium-enriched wheat is added to obtain slurry, and the obtained slurry is used for the subsequent enzymolysis step. In the enzymolysis step, amylase accounting for 0.8 percent of the weight of the selenium-enriched wheat, alkaline protease accounting for 1.5 percent of the weight of the selenium-enriched wheat and aminopeptidase accounting for 0.8 percent of the weight of the selenium-enriched wheat are sequentially added into the slurry, the time interval between the addition of the two adjacent enzymes is 0.6h, the mixture is stirred for enzymolysis, and after the addition of the last enzyme is finished, the mixture is continuously stirred for enzymolysis for 3.5 h; the temperature of stirring and enzymolysis is kept at 57 ℃.
Examples of the experiments
1. Small peptide molecular weight distribution determination
(1) After solid-liquid separation in the examples and the comparative examples, the separated liquid before nanofiltration filtration is dried, and then the molecular weight distribution of the small peptide is measured, and the results are shown in the following table.
TABLE 1 molecular weight distribution of small peptides
| The percentage of peptide molecules with a molecular weight of less than or equal to 1000 Dalton in the total peptide molecule content （wt%） | The total content of dipeptide and tripeptide is the total peptide molecule content （wt%） | The total content of single amino acid accounts for the total peptide molecule content （wt%） | |
| Examples 1 | 92.4 | 56.4 | 7.2 |
| Examples 2 | 93.5 | 55.8 | 6.3 |
| Examples 3 | 91.8 | 57.0 | 7.5 |
| Comparative example 1 | 77.9 | 15.4 | 22.0 |
| Comparative example 2 | 90.0 | 42.8 | 10.0 |
(2) The molecular weight distribution of the small peptide is measured by taking the biological selenium active peptide finally prepared in the example and the comparative example respectively, and the measurement results are shown in the following table.
TABLE 2 molecular weight distribution of small peptides
| The peptide molecules with the molecular weight less than or equal to 1000 daltons account for the total content of the peptide molecules Percent (wt%) | The total content of dipeptide and tripeptide is in total peptide molecules Content (wt%) | The total content of single amino acids in total peptide molecules Content (wt%) | Peptide molecule in biological selenium active peptide Content (wt%) | |
| Practice of Example 1 | 99.3 | 76.7 | 0.6 | 99.9 |
| Practice of Example 2 | 99.6 | 77.7 | 0.3 | 99.8 |
| Practice of Example 3 | 98.9 | 76.6 | 0.9 | 99.6 |
| Comparison of Example 1 | 85.2 | 25.4 | 13.0 | 53.4 |
| Comparison of Example 2 | 98.4 | 62.4 | 1.7 | 90.1 |
From the above, it can be seen that the content percentages of the dipeptide and the tripeptide in the biological selenium active peptide prepared by the invention are high, the content of the single amino acid is low, compared with the content percentages of the dipeptide and the tripeptide in the biological selenium active peptide prepared by the comparative example 1, the content of the single amino acid is high, the biological selenium active peptide with higher content percentages of the dipeptide and the tripeptide can be obtained only by a plurality of filtering steps, the process is complex, the period is long, and the cost is high. The content percentages of the di-and tripeptides in the bio-selenium active peptide prepared in the comparative example 2 were lower than the content percentages of the di-and tripeptides in the bio-selenium active peptide prepared in the present invention.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. A preparation method of biological selenium active peptide is characterized by comprising the following steps:
(1) soaking the selenium-rich plant seeds in water, taking out the selenium-rich plant seeds, spraying water for 2-3 times every day for germination, discharging after 6-7 days of germination to obtain seed germination raw materials, and then stirring to obtain a slurry;
(2) and enzymolysis: sequentially adding amylase accounting for 0.5-0.8 percent of the weight of the seed germination raw material, alkaline protease accounting for 1-1.5 percent of the weight of the seed germination raw material and aminopeptidase accounting for 0.5-0.8 percent of the weight of the seed germination raw material into the slurry, wherein the adding time interval of two adjacent enzymes is 0.4-0.6h, stirring for enzymolysis, and continuously stirring for enzymolysis for 3-3.5h after the last enzyme is added; the temperature of stirring and enzymolysis is kept between 55 and 57 ℃;
(3) enzyme deactivation;
(4) carrying out solid-liquid separation;
(5) filtering the separated liquid obtained in the step (4) by using a nanofiltration membrane of 0.8-1.2nm, reserving the permeate, then filtering by using a nanofiltration membrane of 0.2-0.3nm, and reserving the membrane retentate;
(6) adding organic selenium into the membrane trapped fluid obtained in the step (5) to carry out peptide selenium chelation;
(7) and (4) concentrating and drying the chelating solution obtained in the step (6).
2. The method for preparing bioactive selenium peptide as claimed in claim 1, wherein the selenium-enriched plant seeds are added into purified water 5-6 times the weight of the selenium-enriched plant seeds, and soaked for 12-18 h; and/or
The germination temperature is 15-20 deg.C, and the humidity is 65-70%.
3. The method of claim 1 or 2, wherein the bioactive peptide is selected from the group consisting of,
the enzyme activity of the amylase is 17-23U/mg;
the enzyme activity of the alkaline protease is 25-31U/mg;
the enzyme activity of the aminopeptidase is 22-28U/mg.
4. The method for preparing biological selenium active peptide according to claim 1 or 2, wherein in step (3), the temperature of the enzymatic hydrolysate obtained in step (2) is raised to 90-95 ℃, and the enzymatic hydrolysate is kept for 10-15min to inactivate enzyme.
5. The method for preparing bioactive selenium peptide according to claim 1 or 2, wherein in the step (4), in the solid-liquid separation step, the solid is firstly filtered by a horizontal spiral centrifuge to remove solids, then filtered by a plate-and-frame filter, and finally centrifuged and purified by a three-phase centrifuge;
the rotating speed of the horizontal screw centrifuge is 3600 revolutions per minute;
the aperture of the filter plate in the plate frame filter is 0.3-0.4 μm;
the rotational speed of the three-phase centrifuge is 18000 rpm.
6. The method for preparing bioactive selenium peptide as claimed in claim 1 or 2, wherein in step (6), the addition amount of organic selenium is 0.06-0.07 ‰ of the weight of the seed germination raw material.
7. The method for preparing bioactive peptides as claimed in claim 1 or 2, wherein in step (6), the conditions for selenium chelation of the peptides are as follows: stirring at 45-50 deg.C for 30-45min at 36-45 r/min.
8. The biological selenium active peptide prepared by the preparation method of the biological selenium active peptide in any one of claims 1 to 7, wherein the mass percentage of peptide molecules with the relative molecular weight of less than or equal to 1000 daltons in the peptide molecules of the biological selenium active peptide in the total peptide molecules is more than 98%, and the content of the peptide molecules in the biological selenium active peptide is more than 95 wt%.
9. The bioactive peptides of claim 8 wherein the total dipeptide and tripeptide content of the bioactive peptides is > 45wt% of the total peptide molecule content.
10. Use of the bioactive selenium peptide prepared by the method for preparing the bioactive selenium peptide according to any one of claims 1 to 7 or the bioactive selenium peptide according to claim 8 or 9 in the preparation of feed additives, nutritional formula foods, health products, special medical foods or medicines.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210813744.XA CN114891851B (en) | 2022-07-12 | 2022-07-12 | Biological selenium active peptide and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210813744.XA CN114891851B (en) | 2022-07-12 | 2022-07-12 | Biological selenium active peptide and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114891851A true CN114891851A (en) | 2022-08-12 |
| CN114891851B CN114891851B (en) | 2022-09-27 |
Family
ID=82730400
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210813744.XA Active CN114891851B (en) | 2022-07-12 | 2022-07-12 | Biological selenium active peptide and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114891851B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160235108A1 (en) * | 2013-10-28 | 2016-08-18 | GuiQin Zhang | Selenium-rich wheat whole-sprout juice beverage and method for preparation thereof |
| CN108477376A (en) * | 2018-02-05 | 2018-09-04 | 武汉佳阳绿昶食品有限公司 | A kind of preparation method of selenium-rich and zinc-rich protein peptides |
| CN109576331A (en) * | 2018-12-21 | 2019-04-05 | 河北肽都生物科技有限公司 | A kind of extracting method of wheat germ peptide |
| CN113322295A (en) * | 2020-10-20 | 2021-08-31 | 河北肽丰生物科技有限公司 | Preparation method of selenium-rich triticale malt oligosaccharide peptide and product thereof |
-
2022
- 2022-07-12 CN CN202210813744.XA patent/CN114891851B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160235108A1 (en) * | 2013-10-28 | 2016-08-18 | GuiQin Zhang | Selenium-rich wheat whole-sprout juice beverage and method for preparation thereof |
| CN108477376A (en) * | 2018-02-05 | 2018-09-04 | 武汉佳阳绿昶食品有限公司 | A kind of preparation method of selenium-rich and zinc-rich protein peptides |
| CN109576331A (en) * | 2018-12-21 | 2019-04-05 | 河北肽都生物科技有限公司 | A kind of extracting method of wheat germ peptide |
| CN113322295A (en) * | 2020-10-20 | 2021-08-31 | 河北肽丰生物科技有限公司 | Preparation method of selenium-rich triticale malt oligosaccharide peptide and product thereof |
Non-Patent Citations (1)
| Title |
|---|
| 王杨阳等: "酶法水解富硒平菇蛋白工艺优化", 《食品工业科技》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114891851B (en) | 2022-09-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105586379B (en) | Preparation method of collagen active peptide with cancer cell proliferation inhibition effect | |
| WO2012155295A1 (en) | Industrial production method for producing antihypertensive bioactive peptide | |
| CN1347285A (en) | Highly soluble and stable mineral supplements | |
| CN111685286B (en) | Oyster peptide with blood lipid reducing function and preparation method and application thereof | |
| CN111893157A (en) | Quinoa peptide for reducing blood sugar and preparation method and application thereof | |
| CN102987275B (en) | A kind of preparation method of the soy oligosaccharides glycopeptide of high biological value | |
| CN106538844B (en) | Preparation method of marine organism protein peptide chelated copper | |
| CN100441692C (en) | Chitosan oligosaccharide sulphate and preparation method thereof | |
| CN114158666B (en) | Preparation method and application of soybean oligopeptide | |
| CN102051392A (en) | Chlorhematin and preparation method thereof as well as blood-enriching pharmaceutical composition | |
| CN114891851B (en) | Biological selenium active peptide and preparation method and application thereof | |
| WO2024103619A1 (en) | Preparation method for light-colored water-soluble vegetable protein | |
| CN108201136A (en) | Chromium-rich nutrition fortifier and preparation method thereof | |
| CN105777534B (en) | Calcium citrate salts and preparation method thereof and purposes | |
| CN110897059A (en) | Method for preparing plant hydrolyzed protein beverage by using wheat germs | |
| CN114875102B (en) | Biological iron active peptide and preparation method and application thereof | |
| CN108464442A (en) | A kind of preparation method of the natural selenium-rich food of cytotrophy | |
| CN114875103B (en) | Small peptide chelated calcium and preparation method and application thereof | |
| CN1864545A (en) | Preparation of highland barley SOD an conditioning food containing highland barley SOD | |
| JP2001240599A (en) | Lactic oligosaccharide fraction | |
| KR20050122289A (en) | Manufacturing process of chitosan-selenium complex | |
| CN114410724B (en) | Paeonia ostii peptide with tyrosinase activity inhibiting function and preparation method and application thereof | |
| CN114317656B (en) | Bioactive hairtail peptide microelement chelate and preparation method thereof | |
| CN116694719A (en) | Preparation method and application of small-molecule corn oligopeptide | |
| CN108094671A (en) | A kind of maize oligopeptide preparation process |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |