CN115948491A - Preparation method of selenium peptide chelate for resisting oxidation and preventing tumors - Google Patents
Preparation method of selenium peptide chelate for resisting oxidation and preventing tumors Download PDFInfo
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- CN115948491A CN115948491A CN202310018745.XA CN202310018745A CN115948491A CN 115948491 A CN115948491 A CN 115948491A CN 202310018745 A CN202310018745 A CN 202310018745A CN 115948491 A CN115948491 A CN 115948491A
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- hericium erinaceus
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- 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
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Abstract
The invention relates to a preparation method of a selenium peptide chelate for resisting oxidation and preventing tumors. The silkworm chrysalis and selenium-rich hericium erinaceus are selected, the protein in the silkworm chrysalis and selenium-rich hericium erinaceus is extracted by an alkali-soluble acid precipitation method and an ammonium sulfate precipitation method, the extraction rate is high, the bioactive peptide with high selenium chelating activity is prepared by controlling the enzymolysis time of the protein and independently purifying, the bioactive peptide is chelated with inorganic selenium to prepare a selenium peptide chelate, the obtained chelate contains animal and plant proteins and trace elements, and has the functional activities of oxidation resistance, antibiosis, tumor resistance, immunoregulation and the like.
Description
Technical Field
The invention relates to the technical field of selenium peptide chelates, in particular to a preparation method of a selenium peptide chelate for resisting oxidation and preventing tumors.
Background
Selenium is an important mineral element necessary for human body, the selenium content of human body is closely related to the health condition of human body, the human body mainly obtains selenium by diet, and the artificial selenium supplement method is an oral selenium-containing preparation. Silkworm pupa contains abundant nutrient substances, such as protein, fatty acid, vitamins and the like. Silkworm pupa protein is composed of 18 amino acids, wherein 8 essential amino acids account for 42 percent of the total amount of amino acids, and the silkworm pupa protein is a good biological peptide source. At present, researches find that the silkworm pupa protein source biological peptide has the functional activities of resisting oxidation, reducing blood pressure, reducing blood fat, resisting tumors, regulating immunity and the like. The hericium erinaceus is a fungus which is famous because the shape is exactly like that of the hericium erinaceus, the hericium erinaceus is high in nutritional ingredients, each hundred grams of dry products contain 26.3 grams of protein and 17 kinds of amino acids, wherein the protein accounts for 8 kinds required by a human body, the amino acids are rich in variety and contain rare amino acids which are rare in other foods, and the hericium erinaceus protein has richer biological activity compared with common animal and plant proteins.
The selenium-rich hericium erinaceus and the silkworm chrysalis are deeply processed and chelated by utilizing a biotechnology to prepare the polypeptide-selenium chelate, and the chelate obtained by deep processing contains animal and plant proteins and is rich in trace elements, so that the selenium-rich hericium erinaceus and the silkworm chrysalis have high development value and application prospect.
Disclosure of Invention
In view of the problems in the prior art, the invention discloses a preparation method of a selenium peptide chelate for resisting oxidation and preventing tumors, which comprises the following steps:
step one, preparing silkworm chrysalis peptide powder, which comprises the following steps:
step 11, selecting silkworm chrysalis, cleaning, putting into airflow drying equipment, and continuously drying by using 10-20 ℃ air until dried silkworm chrysalis is obtained;
step 12, crushing the dried silkworm chrysalis, and screening by using a 900-1100-mesh screen to obtain silkworm chrysalis powder;
step 13, mixing the silkworm chrysalis powder with purified water to obtain a suspension with the mass concentration of 2-5 wt%;
step 14, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 9-11, soaking for 20-30min at the temperature of 25-35 ℃, heating the solution to 50-70 ℃, adding alkaline protease accounting for 1-3% of the silkworm chrysalis powder in mass, carrying out enzymolysis at constant temperature for 1-3h, adjusting the pH to 5.0-7.0, adding papain accounting for 0.5-3% of the silkworm chrysalis powder in mass, stirring at constant temperature for hydrolysis for 1-3h, heating to 80-90 ℃, and inactivating the enzyme for 10min;
step 15, cooling the solution after enzyme deactivation to normal temperature, centrifuging for 10min at the speed of 4000r/min by a centrifuge, and collecting supernatant to obtain a silkworm pupa peptide solution;
step 16, carrying out ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and carrying out freeze drying to obtain silkworm pupa peptide freeze-dried powder;
step two, preparing the hericium erinaceus selenoprotein specifically as follows:
step 21, selecting selenium-rich hericium erinaceus, carrying out superfine grinding on the selenium-rich hericium erinaceus, and screening by using a 900-1100-mesh screen to obtain selenium-rich hericium erinaceus powder;
step 22, mixing the selenium-enriched hericium erinaceus powder with purified water, and gradually adding solid ammonium sulfate to ensure that the saturation of the ammonium sulfate reaches 44-58wt%;
step 23, centrifuging by a centrifuge to obtain hericium erinaceus selenoprotein precipitate, and dissolving the hericium erinaceus selenoprotein in water to enable the mass concentration to reach 2-6wt%;
step 24, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 8-10, adding alkaline protease with the mass of 2-5% of hericium erinaceus selenoprotein at the temperature of 30-60 ℃, performing enzymolysis for 1-3h at constant temperature, heating to 80-90 ℃, and inactivating the enzyme for 10min;
step 25, cooling the solution after enzyme deactivation to normal temperature, centrifuging for 10min at the speed of 4000r/min by a centrifugal machine, and collecting supernatant to obtain a hericium erinaceus selenoprotein polypeptide composite solution;
26, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 2000Da, concentrating a penetrating fluid, and performing freeze drying to obtain hericium erinaceus selenoprotein polypeptide freeze-dried powder;
and step three, mixing the silkworm pupa peptide freeze-dried powder obtained in the step one and the hericium erinaceus selenoprotein polypeptide freeze-dried powder obtained in the step two in proportion, and heating at the temperature of 200-400 ℃ for preparation to obtain the silkworm pupa peptide hericium erinaceus protein-selenium chelate.
As a preferred scheme of the invention, in the step three, the ratio of the silkworm pupa peptide freeze-dried powder to the hericium erinaceus selenoprotein polypeptide freeze-dried powder is as follows: the lyophilized powder of the hericium erinaceus selenoprotein = 2-5.
The invention has the beneficial effects that: the silkworm chrysalis and selenium-rich hericium erinaceus are selected, the protein in the silkworm chrysalis and selenium-rich hericium erinaceus is extracted by an alkali-soluble acid precipitation method and an ammonium sulfate precipitation method, the extraction rate is high, the bioactive peptide with high selenium chelating activity is prepared by controlling the enzymolysis time of the protein and independently purifying, the bioactive peptide is chelated with inorganic selenium to prepare a selenium peptide chelate, the obtained chelate contains animal and plant proteins and trace elements, and has the functional activities of oxidation resistance, antibiosis, tumor resistance, immunoregulation and the like.
Detailed Description
Example 1
The invention relates to a preparation method of a selenium peptide chelate for resisting oxidation and preventing tumors, which comprises the following steps:
step one, preparing silkworm chrysalis peptide powder, which specifically comprises the following steps:
step 11, selecting silkworm chrysalis, cleaning, putting into airflow drying equipment, and continuously drying by using 10-DEG C air until dried silkworm chrysalis is obtained;
step 12, crushing the dried silkworm chrysalis, and screening by using a 900-mesh screen to obtain silkworm chrysalis powder;
step 13, mixing the silkworm chrysalis powder with purified water to obtain a suspension with the mass concentration of 2wt%;
step 14, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 9, soaking for 20min at the state of 25 ℃, heating the solution to 50 ℃, adding alkaline protease accounting for 1% of the weight of the silkworm chrysalis powder, carrying out enzymolysis at constant temperature for 1h, adjusting the pH to 5.0, adding papain accounting for 0.5% of the weight of the silkworm chrysalis powder, stirring at constant temperature for hydrolysis for 1h, heating to 80 ℃, and inactivating the enzyme for 10min;
step 15, cooling the solution after enzyme deactivation to normal temperature, centrifuging for 10min at the speed of 4000r/min by a centrifuge, and collecting supernatant to obtain a silkworm pupa peptide solution;
step 16, carrying out ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and carrying out freeze drying to obtain silkworm pupa peptide freeze-dried powder;
step two, preparing the hericium erinaceus selenoprotein concretely as follows:
step 21, selecting selenium-rich hericium erinaceus, carrying out superfine grinding on the selenium-rich hericium erinaceus, and screening by using a 900-mesh screen to obtain selenium-rich hericium erinaceus powder;
step 22, mixing the selenium-enriched hericium erinaceus powder with purified water, and gradually adding solid ammonium sulfate to ensure that the saturation of the ammonium sulfate reaches 44wt%;
step 23, centrifuging by a centrifuge to obtain hericium erinaceus selenoprotein precipitate, and dissolving the hericium erinaceus selenoprotein in water to enable the mass concentration to reach 2wt%;
24, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 8, adding alkaline protease with the mass of 2% of the hericium erinaceus selenoprotein at the temperature of 30 ℃, carrying out enzymolysis for 1 hour at constant temperature, heating to 80 ℃, and inactivating the enzyme for 10 minutes;
step 25, cooling the solution after enzyme deactivation to normal temperature, centrifuging the solution for 10min at the speed of 4000r/min by using a centrifugal machine, and collecting supernatant to obtain a hericium erinaceus selenoprotein polypeptide composite solution;
step 26, performing ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and performing freeze drying to obtain hericium erinaceus selenoprotein polypeptide freeze-dried powder;
step three, mixing the silkworm chrysalis peptide freeze-dried powder obtained in the step one and the hericium erinaceus selenoprotein polypeptide freeze-dried powder obtained in the step two according to the proportion of 2.
Example 2
The invention relates to a preparation method of a selenium peptide chelate for resisting oxidation and preventing tumors, which comprises the following steps:
step one, preparing silkworm chrysalis peptide powder, which comprises the following steps:
step 11, selecting silkworm chrysalis, cleaning, putting into airflow drying equipment, and continuously drying by 105 ℃ air until obtaining dried silkworm chrysalis;
step 12, crushing the dried silkworm chrysalis, and screening by using a 1000-mesh screen to obtain silkworm chrysalis powder;
step 13, mixing the silkworm chrysalis powder with purified water to obtain a suspension with the mass concentration of 4wt%;
step 14, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 10, soaking for 25min at the temperature of 30 ℃, heating the solution to 60 ℃, adding alkaline protease with the mass of 2% of the silkworm chrysalis powder, carrying out enzymolysis at constant temperature for 2h, adjusting the pH to 6.0, adding papain with the mass of 1.5% of the silkworm chrysalis powder, stirring at constant temperature for hydrolysis for 2h, heating to 85 ℃, and inactivating the enzyme for 10min;
step 15, cooling the solution after enzyme deactivation to normal temperature, centrifuging for 10min at the speed of 4000r/min by a centrifuge, and collecting supernatant to obtain a silkworm pupa peptide solution;
step 16, performing ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and performing freeze drying to obtain silkworm chrysalis peptide freeze-dried powder;
step two, preparing the hericium erinaceus selenoprotein specifically as follows:
step 21, selecting selenium-rich hericium erinaceus, carrying out superfine grinding on the selenium-rich hericium erinaceus, and screening by using a 1000-mesh screen to obtain selenium-rich hericium erinaceus powder;
step 22, mixing the selenium-rich hericium erinaceus powder with purified water, and gradually adding solid ammonium sulfate to enable the saturation degree of the ammonium sulfate to reach 50wt%;
step 23, centrifuging by a centrifuge to obtain hericium erinaceus selenoprotein precipitate, and dissolving the hericium erinaceus selenoprotein in water to enable the mass concentration to reach 4wt%;
step 24, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 9, adding alkaline protease with the mass of 4% of the hericium erinaceus selenoprotein at the temperature of 45 ℃, keeping the temperature for 2 hours, heating to 85 ℃, and inactivating the enzyme for 10 minutes; step 25, cooling the solution after enzyme deactivation to normal temperature, centrifuging the solution for 10min at the speed of 4000r/min by using a centrifugal machine, and collecting supernatant to obtain a hericium erinaceus selenoprotein polypeptide composite solution;
step 26, performing ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and performing freeze drying to obtain hericium erinaceus selenoprotein polypeptide freeze-dried powder;
step three, mixing the silkworm chrysalis peptide freeze-dried powder obtained in the step one and the hericium erinaceus selenoprotein polypeptide freeze-dried powder obtained in the step two according to the proportion of 3.
Example 3
The invention relates to a preparation method of a selenium peptide chelate for resisting oxidation and preventing tumors, which comprises the following steps:
step one, preparing silkworm chrysalis peptide powder, which specifically comprises the following steps:
step 11, selecting silkworm chrysalis, cleaning, putting into airflow drying equipment, and continuously drying by air at 20 ℃ until dried silkworm chrysalis is obtained;
step 12, crushing the dried silkworm chrysalis, and screening by using a 1100-mesh screen to obtain silkworm chrysalis powder;
step 13, mixing the silkworm chrysalis powder with purified water to obtain a suspension with the mass concentration of 5 wt%;
step 14, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 11, soaking for 30min at the state of 35 ℃, heating the solution to 70 ℃, adding alkaline protease with the mass being 3% of the silkworm chrysalis powder, carrying out constant-temperature enzymolysis for 3h, adjusting the pH to 7.0, adding papain with the mass being 3% of the silkworm chrysalis powder, carrying out constant-temperature stirring hydrolysis for 3h, heating to 90 ℃, and inactivating the enzyme for 10min;
step 15, cooling the solution after enzyme deactivation to normal temperature, centrifuging the solution for 10min at the speed of 4000r/min by a centrifugal machine, and collecting supernatant to obtain a silkworm pupa peptide solution;
step 16, carrying out ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and carrying out freeze drying to obtain silkworm pupa peptide freeze-dried powder;
step two, preparing the hericium erinaceus selenoprotein specifically as follows:
step 21, selecting selenium-rich hericium erinaceus, carrying out superfine grinding on the selenium-rich hericium erinaceus, and screening by using a 1100-mesh screen to obtain selenium-rich hericium erinaceus powder;
step 22, mixing the selenium-enriched hericium erinaceus powder with purified water, and gradually adding solid ammonium sulfate to enable the saturation degree of the ammonium sulfate to reach 58wt%;
step 23, centrifuging by a centrifuge to obtain hericium erinaceus selenoprotein precipitate, and dissolving the hericium erinaceus selenoprotein in water to make the mass concentration reach 6wt%;
step 24, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 10, adding alkaline protease with the mass of 5% of hericium erinaceus selenoprotein at the temperature of 60 ℃, carrying out enzymolysis at constant temperature for 3 hours, heating to 90 ℃, and inactivating the enzyme for 10min;
step 25, cooling the solution after enzyme deactivation to normal temperature, centrifuging the solution for 10min at the speed of 4000r/min by using a centrifugal machine, and collecting supernatant to obtain a hericium erinaceus selenoprotein polypeptide composite solution;
step 26, performing ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and performing freeze drying to obtain hericium erinaceus selenoprotein polypeptide freeze-dried powder;
step three, mixing the silkworm chrysalis peptide freeze-dried powder obtained in the step one and the hericium erinaceus selenium protein polypeptide freeze-dried powder obtained in the step two according to the proportion of 5, and heating and preparing at the temperature of 400 ℃ to obtain the silkworm chrysalis peptide hericium erinaceus protein-selenium chelate.
Components not described in detail herein are prior art.
Although the present invention has been described in detail with reference to the specific embodiments, the present invention is not limited to the above embodiments, and various changes and modifications without inventive changes may be made within the knowledge of those skilled in the art without departing from the spirit of the present invention.
Claims (2)
1. A preparation method of a selenium peptide chelate for resisting oxidation and preventing tumors is characterized by comprising the following steps:
step one, preparing silkworm chrysalis peptide powder, which specifically comprises the following steps:
step 11, selecting silkworm chrysalis, cleaning, putting into airflow drying equipment, and continuously drying by using 10-20 ℃ air until dried silkworm chrysalis is obtained;
step 12, crushing the dried silkworm chrysalis, and screening by using a 900-1100-mesh screen to obtain silkworm chrysalis powder;
step 13, mixing the silkworm chrysalis powder with purified water to obtain a suspension with the mass concentration of 2-5 wt%;
step 14, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 9-11, soaking for 20-30min at the temperature of 25-35 ℃, heating the solution to 50-70 ℃, adding alkaline protease accounting for 1-3% of the silkworm chrysalis powder in mass, carrying out enzymolysis at constant temperature for 1-3h, adjusting the pH to 5.0-7.0, adding papain accounting for 0.5-3% of the silkworm chrysalis powder in mass, stirring at constant temperature for hydrolysis for 1-3h, heating to 80-90 ℃, and inactivating the enzyme for 10min;
step 15, cooling the solution after enzyme deactivation to normal temperature, centrifuging for 10min at the speed of 4000r/min by a centrifuge, and collecting supernatant to obtain a silkworm pupa peptide solution;
step 16, performing ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and performing freeze drying to obtain silkworm chrysalis peptide freeze-dried powder;
step two, preparing the hericium erinaceus selenoprotein specifically as follows:
step 21, selecting selenium-rich hericium erinaceus, carrying out superfine grinding on the selenium-rich hericium erinaceus, and screening by using a 900-1100-mesh screen to obtain selenium-rich hericium erinaceus powder;
step 22, mixing the selenium-enriched hericium erinaceus powder with purified water, and gradually adding solid ammonium sulfate to ensure that the saturation of the ammonium sulfate reaches 44-58wt%;
step 23, centrifuging by a centrifuge to obtain hericium erinaceus selenoprotein precipitate, and dissolving the hericium erinaceus selenoprotein in water to enable the mass concentration to reach 2-6wt%;
24, adding a sodium hydroxide solution while stirring to adjust the pH of the solution to 8-10, adding alkaline protease with the mass of 2-5% of the hericium erinaceus selenoprotein at the temperature of 30-60 ℃, performing enzymolysis for 1-3 hours at constant temperature, heating to 80-90 ℃, and inactivating the enzyme for 10min;
step 25, cooling the solution after enzyme deactivation to normal temperature, centrifuging for 10min at the speed of 4000r/min by a centrifugal machine, and collecting supernatant to obtain a hericium erinaceus selenoprotein polypeptide composite solution;
step 26, performing ultrafiltration by using an ultrafiltration membrane with the cut-off molecular weight of 2000Da, concentrating a penetrating fluid, and performing freeze drying to obtain hericium erinaceus selenoprotein polypeptide freeze-dried powder;
and step three, mixing the silkworm pupa peptide freeze-dried powder obtained in the step one and the hericium erinaceus selenoprotein polypeptide freeze-dried powder obtained in the step two in proportion, and heating at the temperature of 200-400 ℃ for preparation to obtain the silkworm pupa peptide hericium erinaceus protein-selenium chelate.
2. The method for preparing the selenium peptide chelate complex for resisting oxidation and preventing tumor according to claim 1, which is characterized in that: the ratio of the silkworm chrysalis peptide freeze-dried powder to the hericium erinaceus selenoprotein polypeptide freeze-dried powder in the third step is that the silkworm chrysalis peptide freeze-dried powder: the hericium erinaceus protein polypeptide freeze-dried powder = 2.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117384993A (en) * | 2023-09-15 | 2024-01-12 | 佛山松和宏量健康科技有限公司 | Method for extracting organic selenium from hericium erinaceus |
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2023
- 2023-01-06 CN CN202310018745.XA patent/CN115948491A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117384993A (en) * | 2023-09-15 | 2024-01-12 | 佛山松和宏量健康科技有限公司 | Method for extracting organic selenium from hericium erinaceus |
| CN117384993B (en) * | 2023-09-15 | 2024-05-14 | 佛山松和宏量健康科技有限公司 | Method for extracting organic selenium from hericium erinaceus |
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