CN111349675A - Production process of pseudosciaena crocea oligopeptide - Google Patents
Production process of pseudosciaena crocea oligopeptide Download PDFInfo
- Publication number
- CN111349675A CN111349675A CN201811565754.6A CN201811565754A CN111349675A CN 111349675 A CN111349675 A CN 111349675A CN 201811565754 A CN201811565754 A CN 201811565754A CN 111349675 A CN111349675 A CN 111349675A
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- China
- Prior art keywords
- oligopeptide
- yellow croaker
- large yellow
- enzymolysis
- 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
-
- 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
-
- 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
Abstract
The invention discloses a production process of large yellow croaker oligopeptide, which comprises the following steps: cutting large yellow croaker meat, homogenizing, and performing fine grinding pretreatment to obtain homogenate; adjusting the pH value of the homogenate to be neutral, and adding neutral protease for enzymolysis to obtain primary enzymolysis liquid; adjusting the pH value of the enzymolysis liquid to be alkaline, and adding alkaline protease for enzymolysis to obtain a second enzymolysis liquid; inactivating enzyme of the enzymolysis liquid, filtering, and collecting supernatant; separating the supernatant with ultrafiltration membrane, and collecting the permeate; and adding a polyacrylamide flocculant into the permeate liquid for decoloring and desalting, concentrating in vacuum, and performing spray drying to obtain high-quality pseudosciaena crocea oligopeptide powder. The pseudosciaena crocea oligopeptide is small in molecular weight, has the functions of resisting oxidation and removing free radicals, and can effectively enhance the anti-aging and anti-disease capabilities of a human body.
Description
Technical Field
The invention relates to the technical field of biochemical engineering, in particular to a production process of pseudosciaena crocea oligopeptide.
Background
The oligopeptide is a small molecule active peptide consisting of 2-10 amino acids. The content of active peptide in organism is very small, but it has obvious physiological activity, and each active peptide has its own unique structure, and the different structures determine its unique functional activity. Research proves that the oligopeptide has strong biological activity, such as functions of resisting oxidation, resisting thrombus, reducing blood pressure, resisting fatigue and the like.
China has abundant marine aquatic resources, and the output of mariculture and ocean fishery is steadily increased. The large yellow croaker is also named yellow croaker, king fish, fresh fish, big yellow croaker, red melon, golden dragon, sweet osmanthus yellow croaker, giant Zhongkou, red mouth, chub fish, stone fish and cucumber fish, is one of the traditional four-large marine products (large yellow croaker, small yellow croaker, hairtail and cuttlefish) in China, and is the main economic fish in the offshore region of China. The yellow croaker contains rich protein, trace elements and vitamins, and the rich functional peptide resources of the yellow croaker are not well researched and developed at home. Accordingly, the prior art is yet to be improved and developed.
Disclosure of Invention
The invention aims to solve the technical problem of providing a production process of pseudosciaena crocea oligopeptide.
The technical scheme adopted by the invention for solving the technical problems is that the production process of the large yellow croaker oligopeptide comprises the following steps:
(1) cutting large yellow croaker meat into minced fish, adding water and homogenizing to obtain homogenate;
(2) adjusting the pH value of the homogenate liquid in the step (1) to 6.5-7.0, adding neutral protease accounting for 1-3% of the total mass of protein of the homogenate liquid, stirring and carrying out enzymolysis for 3-6 hours to obtain a primary large yellow croaker enzymolysis liquid;
(3) adjusting the pH of the enzymolysis solution to 7.0-9.0 with NaOH solution, adding alkaline protease in an amount of 1-5% of the total amount of protein, stirring for enzymolysis for 3-6h to obtain second enzymolysis solution of large yellow croaker;
(4) heating the large yellow croaker enzymolysis liquid obtained in the step (3) to 80-95 ℃ to inactivate enzyme for 20-30 min;
(5) filtering the enzyme-killing solution, centrifuging and collecting filtrate;
(6) performing membrane separation on the supernatant collected in the step (4) by using a 1000-2000Da ultrafiltration membrane, and collecting a permeate, namely the pseudosciaena crocea oligopeptide solution;
(7) adding a polyacrylamide flocculant into the large yellow croaker oligopeptide liquid for decoloring;
(8) removing inorganic salt ions from the decolorized solution by an adsorption resin method;
(9) and (4) performing vacuum concentration on the large yellow croaker oligopeptide desalted and decolored solution prepared in the step (8), and performing spray drying to obtain high-quality large yellow croaker oligopeptide powder.
In a further technical scheme, the decoloring step in the step (7) is as follows: using 2-5% of enamine flocculant with the dosage of 5-10% (v/v), stirring for 10-30min, centrifuging, and collecting supernatant.
In a further technical scheme, the adsorption resin in the step (8) is D101 macroporous adsorption resin.
In a further technical scheme, the spray drying conditions in the step (9) are as follows: the temperature is 105-125 ℃.
Compared with the prior art, the invention has the following beneficial effects:
(1) the invention provides a preparation method suitable for industrial production of high-quality pseudosciaena crocea oligopeptide, which is simple in preparation process and reasonable in design and provides a feasible scheme for industrial production of high-quality pseudosciaena crocea oligopeptide.
(2) After the enzymolysis of the large yellow croaker protein by using the neutral protease combined with the alkaline protease, the small peptide with the molecular weight less than 2kDa is separated by an ultrafiltration membrane, and the content of the small peptide is more than 95 percent. Due to small molecular weight, the product has the functions of resisting oxidation and scavenging free radicals, and can effectively enhance the capabilities of human body in resisting aging and diseases.
Detailed Description
For a better understanding of the present invention, reference is made to the following examples. It is to be understood that these examples are for further illustration of the invention and are not intended to limit the scope of the invention. In addition, it should be understood that the invention is not limited to the above-described embodiments, but is capable of various modifications and changes within the scope of the invention.
Example 1
Cutting large yellow croaker meat into minced fish, adding water and homogenizing to obtain homogenate; adjusting the pH value of the homogenate to 7.0, adding neutral protease accounting for 2% of the total mass of protein of the homogenate, and stirring for enzymolysis for 5 hours to obtain a primary large yellow croaker enzymolysis liquid; adjusting the pH of the enzymolysis solution to 8.0 by using NaOH solution, adding alkaline protease with the addition amount of 3% of the total amount of protein, stirring and carrying out enzymolysis for 4h to obtain a second large yellow croaker enzymolysis solution; heating the obtained large yellow croaker enzymolysis liquid to 85 ℃ to inactivate enzyme for 25 min; filtering the enzyme-killing solution, centrifuging and collecting filtrate; performing membrane separation on the collected filtrate by using a 1000-plus 2000Da ultrafiltration membrane, and collecting a permeate, namely the pseudosciaena crocea oligopeptide solution; adding a polyacrylamide flocculant with the concentration of 3% into the large yellow croaker oligopeptide liquid for decoloring; removing inorganic salt ions from the decolorized solution through D101 macroporous adsorption resin; and (3) carrying out vacuum concentration on the prepared large yellow croaker oligopeptide desalted and decolored solution, and then carrying out spray drying to obtain high-quality large yellow croaker oligopeptide powder.
Example 2
Cutting large yellow croaker meat into minced fish, adding water and homogenizing to obtain homogenate; adjusting the pH value of the homogenate to 6.8, adding neutral protease accounting for 3% of the total mass of protein of the homogenate, and stirring for enzymolysis for 4 hours to obtain a primary large yellow croaker enzymolysis liquid; adjusting the pH of the enzymolysis solution to 8.2 by using NaOH solution, adding alkaline protease with the addition amount of 3.5 percent of the total amount of protein, stirring and carrying out enzymolysis for 5 hours to obtain a second large yellow croaker enzymolysis solution; heating the obtained large yellow croaker enzymolysis liquid to 95 ℃ to inactivate enzyme for 20 min; filtering the enzyme-killing solution, centrifuging and collecting filtrate; performing membrane separation on the collected filtrate by using a 1000-plus 2000Da ultrafiltration membrane, and collecting a permeate, namely the pseudosciaena crocea oligopeptide solution; adding a polyacrylamide flocculant with the concentration of 5% into the large yellow croaker oligopeptide liquid for decoloring; removing inorganic salt ions from the decolorized solution through D101 macroporous adsorption resin; and (3) carrying out vacuum concentration on the prepared large yellow croaker oligopeptide desalted and decolored solution, and then carrying out spray drying to obtain high-quality large yellow croaker oligopeptide powder.
As described above, the present invention can be preferably implemented.
Claims (4)
1. A production process of large yellow croaker oligopeptide is characterized by comprising the following steps:
(1) cutting large yellow croaker meat into minced fish, adding water and homogenizing to obtain homogenate;
(2) adjusting the pH value of the homogenate liquid in the step (1) to 6.5-7.0, adding neutral protease accounting for 1-3% of the total mass of protein of the homogenate liquid, stirring and carrying out enzymolysis for 3-6 hours to obtain a primary large yellow croaker enzymolysis liquid;
(3) adjusting the pH of the enzymolysis solution to 7.0-9.0 with NaOH solution, adding alkaline protease in an amount of 1-5% of the total amount of protein, stirring for enzymolysis for 3-6h to obtain second enzymolysis solution of large yellow croaker;
(4) heating the large yellow croaker enzymolysis liquid obtained in the step (3) to 80-95 ℃ to inactivate enzyme for 20-30 min;
(5) filtering the enzyme-killing solution, centrifuging and collecting filtrate;
(6) performing membrane separation on the supernatant collected in the step (4) by using a 1000-2000Da ultrafiltration membrane, and collecting a permeate, namely the pseudosciaena crocea oligopeptide solution;
(7) adding a polyacrylamide flocculant into the large yellow croaker oligopeptide liquid for decoloring;
(8) removing inorganic salt ions from the decolorized solution by an adsorption resin method;
(9) and (4) performing vacuum concentration on the large yellow croaker oligopeptide desalted and decolored solution prepared in the step (8), and performing spray drying to obtain high-quality large yellow croaker oligopeptide powder.
2. The process for producing pseudosciaena crocea oligopeptide according to claim 1, wherein the process comprises the following steps: the decoloring step in the step (7) is as follows: using 2-5% of enamine flocculant with the dosage of 5-10% (v/v), stirring for 10-30min, centrifuging, and collecting supernatant.
3. The production process of large yellow croaker oligopeptide according to claim 1, wherein the adsorbent resin in step (8) is a D101 macroporous adsorbent resin.
4. The method for producing pseudosciaena crocea oligopeptide according to claim 1, wherein the spray drying conditions in the step (9) are as follows: the temperature is 105-125 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201811565754.6A CN111349675A (en) | 2018-12-20 | 2018-12-20 | Production process of pseudosciaena crocea oligopeptide |
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| CN201811565754.6A CN111349675A (en) | 2018-12-20 | 2018-12-20 | Production process of pseudosciaena crocea oligopeptide |
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| CN111349675A true CN111349675A (en) | 2020-06-30 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114146219A (en) * | 2021-12-29 | 2022-03-08 | 上海璞聚生物科技有限公司 | Soft tissue filler and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114146219A (en) * | 2021-12-29 | 2022-03-08 | 上海璞聚生物科技有限公司 | Soft tissue filler and preparation method thereof |
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Application publication date: 20200630 |
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