CN114395603A - Method for preparing small molecular peptide by hydrolyzing oyster protein with complex enzyme - Google Patents
Method for preparing small molecular peptide by hydrolyzing oyster protein with complex enzyme Download PDFInfo
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- CN114395603A CN114395603A CN202210128309.3A CN202210128309A CN114395603A CN 114395603 A CN114395603 A CN 114395603A CN 202210128309 A CN202210128309 A CN 202210128309A CN 114395603 A CN114395603 A CN 114395603A
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Abstract
The invention discloses a method for preparing micromolecular peptide by hydrolyzing oyster protein with complex enzyme, which comprises the following steps of firstly removing shells of oysters, taking out and cleaning the oyster meat, stirring and crushing the cleaned oyster meat by using crushing equipment to obtain meat paste, adding water which is 5-10% of the weight of the meat paste into the obtained meat paste to finish primary enzymolysis, adding neutral protease which is 0.06-0.1% of the weight of the meat paste into the meat paste obtained after the primary enzymolysis to finish secondary enzymolysis, adding pepsin which is 0.05-0.07% of the weight of the meat paste into the meat paste obtained after the secondary enzymolysis to obtain enzymolysis liquid, and then carrying out spray drying or freeze drying to obtain the micromolecular peptide product of the oysters, and has the beneficial effects that: according to the invention, the oyster meat is crushed into meat paste, so that the mixing effect of the meat paste and the protease is conveniently increased, and during enzymolysis, the three times of enzymolysis of alkaline protease, neutral protease and pepsin are sequentially carried out, so that the molecular weight of molecular peptides in the meat paste is reduced, and the meat paste is convenient for absorption and utilization of a human body.
Description
Technical Field
The invention relates to the technical field of preparation of small molecular peptides, and particularly relates to a method for preparing small molecular peptides by hydrolyzing oyster proteins with complex enzyme.
Background
The Concha Ostreae is a marine product shellfish. Oyster belongs to the phylum mollusca, the class of lamellibranchia, the order of heterophylly column, the superfamily of oyster, the family of oyster, and is a marine product of bivalve shell for both food and medicine. The oyster is cultivated in the south and north China and coastal areas, particularly develops rapidly in the past few years in Guangdong province, the oyster meat is milky, tender and rich in nutrition, and the oyster meat contains rich proteins, vitamins and saccharides, and also contains more than ten amino acids and mineral substance nutrients necessary for human bodies.
Recent studies show that: the oyster contains 18 amino acids, glycogen, B vitamins, taurine, calcium, phosphorus, iron, zinc and other nutrients, and can improve the immunity of the organism after being eaten frequently, and the taurine contained in the oyster can reduce the blood fat and the blood pressure. Clinically, oysters have outstanding curative effects on insomnia, chronic otitis media, infantile hyperhidrosis, hysteromyoma, infantile enuresis and the like. The Concha Ostreae extract has antibacterial effect, and has effects in inhibiting poliovirus and influenza virus, and its water soluble component can improve animal immunity.
In chinese patent No. CN1680578A, a preparation method of oyster active peptide is disclosed, which uses pacific oyster as raw material, hydrolyzes oyster protein by protease, and then desalts and concentrates by nanofiltration technology to obtain oyster active peptide. The method uses protease, the selection of the enzyme is the key for producing active peptide, the hydrolysis capability of the enzyme has specificity, if polypeptide with unique activity is desired to be obtained, the effect is not good when only one enzyme is used, and the molecular weight of the protein peptide prepared by the selected enzyme combination is relatively large.
An effective solution to the problems in the related art has not been proposed yet.
Disclosure of Invention
Aiming at the problems in the related art, the invention provides a method for preparing small molecular peptides by hydrolyzing oyster proteins with complex enzyme, so as to overcome the technical problems in the prior related art.
The technical scheme of the invention is realized as follows:
a method for preparing small molecular peptide by hydrolyzing oyster protein with complex enzyme specifically comprises the following preparation steps:
(1) removing shells of oysters, taking out oyster meat, cleaning, and crushing the cleaned oyster meat by using crushing equipment to obtain meat paste;
(2) adding water which is 5-10% of the weight of the meat paste into the obtained meat paste, adding alkaline protease which is 0.05-0.1% of the weight of the meat paste, mixing, adjusting the pH value to 7-8, performing enzymolysis for 30-40 minutes, taking out the meat paste, and completing the first enzymolysis;
(3) adding neutral protease which is 0.06-0.1% of the weight of the meat paste into the meat paste obtained after the first enzymolysis, keeping the temperature at 45-50 ℃, adjusting the pH value to 6-7, standing for 30-40 minutes, and finishing the second enzymolysis;
(4) adding pepsin which is 0.05-0.07 percent of the weight of the meat paste into the meat paste after the second enzymolysis, heating to 70-85 ℃, homogenizing for 100-140 minutes, and inactivating enzyme for 20 minutes to obtain an enzymolysis liquid;
(5) filtering the enzymolysis solution, adding activated carbon accounting for 1.5-2.5% of the weight of the filtrate, and carrying out decolorization treatment to obtain a small molecular peptide solution;
(6) and (3) carrying out vacuum concentration on the small molecular peptide liquid at 50-60 ℃ to 20% of the initial weight, and then carrying out spray drying or freeze drying to obtain the oyster small molecular peptide product.
Further, the crushing equipment in the step (1) adopts a household crusher, and the rotating speed of the household crusher is 1500-2000 revolutions per minute.
Further, the alkaline protease in the step (2) is one of papain produced by plants and mold protease produced by microorganisms.
Further, the neutral protease in the step (3) is obtained by fermenting and extracting bacillus subtilis, and can be used for various protein hydrolysis treatments. The product can hydrolyze macromolecular protein into amino acids.
Further, the homogenizing pressure in the step (4) is 16-21 Mpa.
The invention provides a method for preparing small molecular peptide by hydrolyzing oyster protein with complex enzyme, which has the following beneficial effects:
(1) the oyster meat is crushed into meat paste, so that the mixing effect of the meat paste and the protease is conveniently increased, and during enzymolysis, the three times of enzymolysis of alkaline protease, neutral protease and pepsin are sequentially performed, so that the molecular weight of molecular peptides in the meat paste is reduced, and the meat paste is convenient for human bodies to absorb and utilize.
(2) And (2) adopting a household grinder as the grinding equipment in the step (1), wherein the rotating speed is 1500-2000 revolutions per minute, so that the oyster meat is thoroughly ground, and the meat pieces are prevented from being produced.
(3) The alkaline protease in the step (2) is one of papain produced by plants and mold protease produced by microorganisms.
(4) The neutral protease in the step (3) is obtained by fermenting and extracting bacillus subtilis, and can be used for various protein hydrolysis treatments.
(5) And (5) in the step (4), the homogenizing pressure is 16-21 Mpa, and the stability of the product is ensured.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a flow chart of a method for preparing small molecule peptides by hydrolyzing oyster proteins with complex enzyme according to an embodiment of the present invention.
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 that can be derived by one of ordinary skill in the art from the embodiments given herein are intended to be within the scope of the present invention.
The invention is further described with reference to the following drawings and detailed description:
the first embodiment is as follows:
referring to fig. 1, a method for preparing small molecule peptides by hydrolyzing oyster proteins with complex enzyme according to an embodiment of the present invention, 1, specifically includes the following steps:
(1) removing shells of oysters, taking out oyster meat, cleaning, and crushing the cleaned oyster meat by using crushing equipment to obtain meat paste;
(2) adding water which is 5-10% of the weight of the meat paste into the obtained meat paste, adding alkaline protease which is 0.05-0.1% of the weight of the meat paste, mixing, adjusting the pH value to 7-8, performing enzymolysis for 30-40 minutes, taking out the meat paste, and completing the first enzymolysis;
(3) adding neutral protease which is 0.06-0.1% of the weight of the meat paste into the meat paste obtained after the first enzymolysis, keeping the temperature at 45-50 ℃, adjusting the pH value to 6-7, standing for 30-40 minutes, and finishing the second enzymolysis;
(4) adding pepsin which is 0.05-0.07 percent of the weight of the meat paste into the meat paste after the second enzymolysis, heating to 70-85 ℃, homogenizing for 100-140 minutes, and inactivating enzyme for 20 minutes to obtain an enzymolysis liquid;
(5) filtering the enzymolysis solution, adding activated carbon accounting for 1.5-2.5% of the weight of the filtrate, and carrying out decolorization treatment to obtain a small molecular peptide solution;
(6) and (3) carrying out vacuum concentration on the small molecular peptide liquid at 50-60 ℃ to 20% of the initial weight, and then carrying out spray drying or freeze drying to obtain the oyster small molecular peptide product.
Example two:
as shown in fig. 1, the method specifically comprises the following preparation steps:
(1) removing shells of oysters, taking out oyster meat, cleaning, and crushing the cleaned oyster meat by using crushing equipment to obtain meat paste;
(2) adding water which is 5 percent of the weight of the meat paste into the obtained meat paste, adding alkaline protease which is 0.05 percent of the weight of the meat paste, mixing, adjusting the pH value to 7, performing enzymolysis for 30 minutes, taking out the meat paste, and completing the first enzymolysis;
(3) adding neutral protease which is 0.06 percent of the weight of the meat paste into the meat paste obtained after the first enzymolysis, keeping the temperature at 45 ℃, adjusting the PH value to 6, standing for 30 minutes, and finishing the second enzymolysis;
(4) adding pepsin which is 0.05 percent of the weight of the meat paste into the meat paste after the second enzymolysis, heating to 70 ℃, homogenizing for 100 minutes, and inactivating enzymes for 20 minutes to obtain an enzymolysis liquid;
(5) filtering the enzymolysis solution, adding activated carbon accounting for 1.5% of the weight of the filtrate, and decolorizing to obtain small molecular peptide solution;
(6) and (3) concentrating the small molecular peptide liquid at 50 ℃ in vacuum to 20% of the initial weight, and then carrying out spray drying or freeze drying to obtain the oyster small molecular peptide product.
Example three:
as shown in fig. 1, the method specifically comprises the following preparation steps:
(1) removing shells of oysters, taking out oyster meat, cleaning, and crushing the cleaned oyster meat by using crushing equipment to obtain meat paste;
(2) adding water which is 10 percent of the weight of the meat paste into the obtained meat paste, adding alkaline protease which is 0.1 percent of the weight of the meat paste, mixing, adjusting the pH value to 8, performing enzymolysis for 40 minutes, taking out the meat paste, and completing the first enzymolysis;
(3) adding neutral protease which is 0.1 percent of the weight of the meat paste into the meat paste obtained after the first enzymolysis, keeping the temperature at 50 ℃, adjusting the pH value to 7, standing for 40 minutes, and finishing the second enzymolysis;
(4) adding pepsin which is 0.07 percent of the weight of the meat paste to the meat paste after the second enzymolysis, heating to 85 ℃, homogenizing for 140 minutes, and inactivating enzymes for 20 minutes to obtain an enzymolysis liquid;
(5) filtering the enzymolysis solution, adding activated carbon with the weight of 2.5% of the filtrate, and decolorizing to obtain small molecular peptide solution;
(6) and (3) concentrating the small molecular peptide liquid at 60 ℃ in vacuum to 20% of the initial weight, and then carrying out spray drying or freeze drying to obtain the oyster small molecular peptide product.
For the convenience of understanding the technical solutions of the present invention, the following detailed description will be made on the working principle or the operation mode of the present invention in the practical process.
In practical application, firstly, shells of oysters are removed, the oyster meat is taken out and cleaned, the cleaned oyster meat is smashed by a smashing device to obtain meat paste, water which is 5-10% of the weight of the meat paste is added into the meat paste, alkaline protease which is 0.05-0.1% of the weight of the meat paste is added into the meat paste and mixed, the pH value of the meat paste is adjusted to 7-8, the meat paste is taken out after 30-40 minutes of enzymolysis, the first enzymolysis is completed, neutral protease which is 0.06-0.1% of the weight of the meat paste is added into the meat paste obtained after the first enzymolysis, the temperature is kept at 45-50 ℃, the pH value of the meat paste is adjusted to 6-7, the meat paste is kept still for 30-40 minutes, the second enzymolysis is completed, pepsin which is 0.05-0.07% of the weight of the meat paste is added into the meat paste obtained after the second enzymolysis, the temperature is increased to 70-85 ℃, the temperature is 100-140 minutes, and the enzyme is inactivated for 20 minutes, the preparation method comprises the steps of obtaining an enzymolysis solution, filtering the enzymolysis solution, adding activated carbon accounting for 1.5-2.5% of the weight of the filtrate into the enzymolysis solution, carrying out decoloring treatment to obtain a small molecular peptide solution, carrying out vacuum concentration on the small molecular peptide solution at 50-60 ℃ to 20% of the initial weight, and carrying out spray drying or freeze drying to obtain the oyster small molecular peptide product.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (5)
1. A method for preparing small molecular peptides by hydrolyzing oyster proteins with complex enzyme is characterized by comprising the following preparation steps:
(1) removing shells of oysters, taking out oyster meat, cleaning, and crushing the cleaned oyster meat by using crushing equipment to obtain meat paste;
(2) adding water which is 5-10% of the weight of the meat paste into the obtained meat paste, adding alkaline protease which is 0.05-0.1% of the weight of the meat paste, mixing, adjusting the pH value to 7-8, performing enzymolysis for 30-40 minutes, taking out the meat paste, and completing the first enzymolysis;
(3) adding neutral protease which is 0.06-0.1% of the weight of the meat paste into the meat paste obtained after the first enzymolysis, keeping the temperature at 45-50 ℃, adjusting the pH value to 6-7, standing for 30-40 minutes, and finishing the second enzymolysis;
(4) adding pepsin which is 0.05-0.07 percent of the weight of the meat paste into the meat paste after the second enzymolysis, heating to 70-85 ℃, homogenizing for 100-140 minutes, and inactivating enzymes for 20 minutes to obtain an enzymolysis liquid;
(5) filtering the enzymolysis solution, adding activated carbon accounting for 1.5-2.5% of the weight of the filtrate, and carrying out decolorization treatment to obtain a small molecular peptide solution;
(6) and (3) carrying out vacuum concentration on the small molecular peptide liquid at 50-60 ℃ to 20% of the initial weight, and then carrying out spray drying or freeze drying to obtain the oyster small molecular peptide product.
2. The method for preparing small molecular peptide by hydrolyzing oyster protein with complex enzyme according to claim 1, wherein the crushing device in the step (1) adopts a household crusher, and the rotating speed of the household crusher is 1500-2000 revolutions per minute.
3. The method for preparing small molecule peptide by hydrolyzing oyster protein with complex enzyme according to claim 1, wherein the alkaline protease in step (2) is one of papain produced by plants and mold protease produced by microorganisms.
4. The method for preparing small molecular peptide by hydrolyzing oyster protein with complex enzyme according to claim 1, wherein the neutral protease in the step (3) is obtained by fermenting and extracting bacillus subtilis, and can be used for various proteolysis treatments. The product can hydrolyze macromolecular protein into amino acids.
5. The method for preparing the small molecular peptide by hydrolyzing the oyster protein with the complex enzyme according to claim 1, wherein the homogenizing pressure in the step (4) is 16-21 Mpa.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114717287A (en) * | 2022-05-07 | 2022-07-08 | 广东还珠海洋生物科技有限公司 | Process for extracting peptide from shellfish |
| CN114794339A (en) * | 2022-05-07 | 2022-07-29 | 广东还珠海洋生物科技有限公司 | Fresh oyster beverage |
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| CN105779540A (en) * | 2016-03-30 | 2016-07-20 | 蔡庭守 | Blueberry small molecular peptide composition, as well as extracting method and application thereof |
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| WO2021142880A1 (en) * | 2020-01-16 | 2021-07-22 | 美国琛蓝营养制品股份有限公司 | Method for producing clam active peptide |
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| CN113957113A (en) * | 2021-11-29 | 2022-01-21 | 海南华研胶原科技股份有限公司 | Oyster oligopeptide and preparation method thereof |
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2022
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Patent Citations (6)
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| CN105695548A (en) * | 2016-03-30 | 2016-06-22 | 蔡庭守 | Preparation method of donkey-hide gelatin small molecular peptide |
| CN105779540A (en) * | 2016-03-30 | 2016-07-20 | 蔡庭守 | Blueberry small molecular peptide composition, as well as extracting method and application thereof |
| CN109825545A (en) * | 2019-04-17 | 2019-05-31 | 微肽科技(海南)有限公司 | The preparation method of one seed oyster oligopeptide freeze-dried powder |
| WO2021142880A1 (en) * | 2020-01-16 | 2021-07-22 | 美国琛蓝营养制品股份有限公司 | Method for producing clam active peptide |
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| CN113957113A (en) * | 2021-11-29 | 2022-01-21 | 海南华研胶原科技股份有限公司 | Oyster oligopeptide and preparation method thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114717287A (en) * | 2022-05-07 | 2022-07-08 | 广东还珠海洋生物科技有限公司 | Process for extracting peptide from shellfish |
| CN114794339A (en) * | 2022-05-07 | 2022-07-29 | 广东还珠海洋生物科技有限公司 | Fresh oyster beverage |
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