CN112724245A - Method for extracting micromolecular collagen peptide by fish scale enzymolysis - Google Patents
Method for extracting micromolecular collagen peptide by fish scale enzymolysis Download PDFInfo
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- 241000251468 Actinopterygii Species 0.000 title claims abstract description 97
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 50
- 102000008186 Collagen Human genes 0.000 title claims abstract description 48
- 108010035532 Collagen Proteins 0.000 title claims abstract description 48
- 229920001436 collagen Polymers 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 22
- 102000004190 Enzymes Human genes 0.000 claims abstract description 35
- 108090000790 Enzymes Proteins 0.000 claims abstract description 35
- 102000029816 Collagenase Human genes 0.000 claims abstract description 24
- 108060005980 Collagenase Proteins 0.000 claims abstract description 24
- 238000002360 preparation method Methods 0.000 claims abstract description 23
- 238000001914 filtration Methods 0.000 claims abstract description 22
- 238000003756 stirring Methods 0.000 claims abstract description 19
- 239000000843 powder Substances 0.000 claims abstract description 17
- 239000007788 liquid Substances 0.000 claims abstract description 16
- 238000000227 grinding Methods 0.000 claims abstract description 11
- 238000010411 cooking Methods 0.000 claims abstract description 6
- 239000012528 membrane Substances 0.000 claims abstract description 6
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 6
- 238000004140 cleaning Methods 0.000 claims description 41
- 229940088598 enzyme Drugs 0.000 claims description 32
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 30
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 238000006243 chemical reaction Methods 0.000 claims description 15
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 15
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 15
- 108090000145 Bacillolysin Proteins 0.000 claims description 11
- 102000035092 Neutral proteases Human genes 0.000 claims description 11
- 108091005507 Neutral proteases Proteins 0.000 claims description 11
- 108090000526 Papain Proteins 0.000 claims description 11
- 102000057297 Pepsin A Human genes 0.000 claims description 11
- 108090000284 Pepsin A Proteins 0.000 claims description 11
- 239000004365 Protease Substances 0.000 claims description 11
- 102000004142 Trypsin Human genes 0.000 claims description 11
- 108090000631 Trypsin Proteins 0.000 claims description 11
- 108010007119 flavourzyme Proteins 0.000 claims description 11
- 235000019834 papain Nutrition 0.000 claims description 11
- 229940055729 papain Drugs 0.000 claims description 11
- 229940111202 pepsin Drugs 0.000 claims description 11
- 239000012588 trypsin Substances 0.000 claims description 11
- 239000003513 alkali Substances 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 10
- 239000000413 hydrolysate Substances 0.000 claims description 10
- 238000000605 extraction Methods 0.000 claims description 9
- 238000005273 aeration Methods 0.000 claims description 8
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 5
- 230000009849 deactivation Effects 0.000 claims description 5
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- 239000012535 impurity Substances 0.000 claims description 5
- 150000002632 lipids Chemical class 0.000 claims description 5
- 210000003097 mucus Anatomy 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 5
- 238000001694 spray drying Methods 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 150000003384 small molecules Chemical class 0.000 claims 8
- 230000000694 effects Effects 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 108050001049 Extracellular proteins Proteins 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
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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
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- Chemical & Material Sciences (AREA)
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- Health & Medical Sciences (AREA)
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- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
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- Microbiology (AREA)
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Abstract
The invention discloses a method for extracting micromolecule collagen peptide by fish scale enzymolysis, which comprises the following steps: adding an enzyme preparation which accounts for 3-8% of the weight of the fed fish scales, and carrying out enzymolysis on the pretreated fish scales to obtain primary collagen peptidase hydrolyzed solution; grinding the filtered fish scales into powder, and then adding an enzyme preparation which is 3% of the weight of the fed fish scales into the powder to obtain secondary collagen peptidase hydrolyzed solution; and stirring and mixing the obtained primary collagen peptidase hydrolyzed solution and the secondary collagen peptidase hydrolyzed solution, then performing fine filtration by using an ultrafiltration membrane, and performing cooking concentration on the collagen peptide liquid after the fine filtration to obtain a collagen peptide concentrated solution. Through carrying out the secondary to the fish scale and drawing, can improve the utilization ratio of collagen peptide in the fish scale, reduce extravagantly, and carry out the enzymolysis to the fish scale through multiple enzyme synergism, can improve the enzymolysis efficiency of fish scale.
Description
Technical Field
The invention belongs to the technical field of collagen peptide extraction, and particularly relates to a method for extracting micromolecule collagen peptide by fish scale enzymolysis.
Background
Collagen peptide is an extracellular protein. Protein peptide consisting of two or more amino acids. The absorption of human body is carried out in a peptide mode, and the absorption utilization rate of the edible protein peptide can reach 100%. Collagen is the most important component of the extracellular matrix.
At present, when fish scale enzymolysis is used for extracting collagen peptide in the prior art, most of the collagen peptide in the fish scale is not extracted, so that waste is caused, and the enzymolysis efficiency is low during extraction.
Disclosure of Invention
The invention aims to provide a method for extracting micromolecule collagen peptide by fish scale enzymolysis, which aims to solve the problems that most of collagen peptide in fish scales is not extracted, waste is caused, and the enzymolysis efficiency is low during extraction.
In order to achieve the purpose, the invention provides the following technical scheme: a method for extracting micromolecular collagen peptide by fish scale enzymolysis comprises the following steps:
the method comprises the following steps: fish scale cleaning
The collected fish scales are poured into cleaning equipment for cleaning, so that impurities adhered to the surfaces of the fish scales can be cleaned;
step two: pretreatment of
Putting the cleaned fish scales into a reaction kettle, adding an alkali solution into the reaction kettle to enable the pH value of the solution in the reaction kettle to be 7-8, stirring for 1-2 hours to remove mucus on the surfaces of the fish scales, adding a sodium bicarbonate solution to remove lipid substances in the fish scales, and cleaning with deionized water;
step three: extracting by enzymolysis
Adding an enzyme preparation which is 3-8% of the weight of the fed fish scales, carrying out enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, then adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, carrying out enzyme deactivation, and filtering the enzymolysis liquid to obtain primary collagen peptidase hydrolysate;
step four: secondary enzymolysis extraction
Grinding the fish scales filtered out in the third step into powder, adding an enzyme preparation which is 3% of the weight of the added fish scales, performing enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, deactivating enzyme, and filtering the enzymolysis liquid to obtain secondary collagen peptidase hydrolysate;
step five: purifying and concentrating
Stirring and mixing the primary collagen peptidase hydrolyzed solution and the secondary collagen peptidase hydrolyzed solution obtained in the third step and the fourth step, then performing fine filtration by using an ultrafiltration membrane, and performing cooking concentration on the collagen peptide liquid after the fine filtration to obtain a collagen peptide concentrated solution;
step six: drying
And (3) rapidly drying the collagen peptide concentrated solution by utilizing spray drying to obtain a finished collagen peptide powder product.
Preferably, the cleaning device in the first step is stirring cleaning and aeration cleaning.
Preferably, the alkali solution in the second step is sodium hydroxide or potassium hydroxide solution.
Preferably, in the fourth step, the fish scales are ground into powder by a fish scale grinding machine.
Preferably, the enzyme preparation in the third step and the fourth step comprises 3-8 parts of neutral protease, 8-14 parts of papain, 10-16 parts of pepsin, 5-7 parts of trypsin and 3-5 parts of flavourzyme.
Preferably, the enzyme preparation comprises 3 parts of neutral protease, 8 parts of papain, 10 parts of pepsin, 5 parts of trypsin and 3 parts of flavourzyme.
Preferably, the enzyme preparation comprises 3-8 parts of neutral protease, 11 parts of papain, 13 parts of pepsin, 6 parts of trypsin and 4 parts of flavourzyme.
Preferably, the enzyme preparation comprises 8 parts of neutral protease, 14 parts of papain, 16 parts of pepsin, 7 parts of trypsin and 5 parts of flavourzyme.
Compared with the prior art, the invention has the beneficial effects that: through carrying out the secondary to the fish scale and drawing, can improve the utilization ratio of collagen peptide in the fish scale, reduce extravagantly, and carry out the enzymolysis to the fish scale through multiple enzyme synergism, can improve the enzymolysis efficiency of fish scale.
Detailed Description
Example 1
A method for extracting micromolecular collagen peptide by fish scale enzymolysis comprises the following steps:
the method comprises the following steps: fish scale cleaning
The collected fish scales are poured into cleaning equipment for cleaning, so that impurities adhered to the surfaces of the fish scales can be cleaned;
step two: pretreatment of
Putting the cleaned fish scales into a reaction kettle, adding an alkali solution into the reaction kettle to enable the pH value of the solution in the reaction kettle to be 7-8, stirring for 1-2 hours to remove mucus on the surfaces of the fish scales, adding a sodium bicarbonate solution to remove lipid substances in the fish scales, and cleaning with deionized water;
step three: extracting by enzymolysis
Adding an enzyme preparation which is 3-8% of the weight of the fed fish scales, carrying out enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, then adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, carrying out enzyme deactivation, and filtering the enzymolysis liquid to obtain primary collagen peptidase hydrolysate;
step four: secondary enzymolysis extraction
Grinding the fish scales filtered out in the third step into powder, adding an enzyme preparation which is 3% of the weight of the added fish scales, performing enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, deactivating enzyme, and filtering the enzymolysis liquid to obtain secondary collagen peptidase hydrolysate;
step five: purifying and concentrating
Stirring and mixing the primary collagen peptidase hydrolyzed solution and the secondary collagen peptidase hydrolyzed solution obtained in the third step and the fourth step, then performing fine filtration by using an ultrafiltration membrane, and performing cooking concentration on the collagen peptide liquid after the fine filtration to obtain a collagen peptide concentrated solution;
step six: drying
And (3) rapidly drying the collagen peptide concentrated solution by utilizing spray drying to obtain a finished collagen peptide powder product.
And in the step one, the cleaning equipment is used for stirring cleaning and aeration cleaning, and the scales can be dispersed during cleaning by utilizing stirring and aeration, so that the phenomenon that the scales are adhered together to influence the cleaning effect is avoided.
And in the second step, the alkali solution is sodium hydroxide or potassium hydroxide solution.
And in the fourth step, the fish scales are grinded into powder by a fish scale grinding machine.
The enzyme preparation in the third step and the fourth step comprises 3 parts of neutral protease, 8 parts of papain, 10 parts of pepsin, 5 parts of trypsin and 3 parts of flavourzyme.
Example 2
A method for extracting micromolecular collagen peptide by fish scale enzymolysis comprises the following steps:
the method comprises the following steps: fish scale cleaning
The collected fish scales are poured into cleaning equipment for cleaning, so that impurities adhered to the surfaces of the fish scales can be cleaned;
step two: pretreatment of
Putting the cleaned fish scales into a reaction kettle, adding an alkali solution into the reaction kettle to enable the pH value of the solution in the reaction kettle to be 7-8, stirring for 1-2 hours to remove mucus on the surfaces of the fish scales, adding a sodium bicarbonate solution to remove lipid substances in the fish scales, and cleaning with deionized water;
step three: extracting by enzymolysis
Adding an enzyme preparation which is 3-8% of the weight of the fed fish scales, carrying out enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, then adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, carrying out enzyme deactivation, and filtering the enzymolysis liquid to obtain primary collagen peptidase hydrolysate;
step four: secondary enzymolysis extraction
Grinding the fish scales filtered out in the third step into powder, adding an enzyme preparation which is 3% of the weight of the added fish scales, performing enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, deactivating enzyme, and filtering the enzymolysis liquid to obtain secondary collagen peptidase hydrolysate;
step five: purifying and concentrating
Stirring and mixing the primary collagen peptidase hydrolyzed solution and the secondary collagen peptidase hydrolyzed solution obtained in the third step and the fourth step, then performing fine filtration by using an ultrafiltration membrane, and performing cooking concentration on the collagen peptide liquid after the fine filtration to obtain a collagen peptide concentrated solution;
step six: drying
And (3) rapidly drying the collagen peptide concentrated solution by utilizing spray drying to obtain a finished collagen peptide powder product.
And in the step one, the cleaning equipment is used for stirring cleaning and aeration cleaning, and the scales can be dispersed during cleaning by utilizing stirring and aeration, so that the phenomenon that the scales are adhered together to influence the cleaning effect is avoided.
And in the second step, the alkali solution is sodium hydroxide or potassium hydroxide solution.
And in the fourth step, the fish scales are grinded into powder by a fish scale grinding machine.
The enzyme preparation in the third step and the fourth step comprises 3-8 parts of neutral protease, 11 parts of papain, 13 parts of pepsin, 6 parts of trypsin and 4 parts of flavourzyme.
Embodiment 3
A method for extracting micromolecular collagen peptide by fish scale enzymolysis comprises the following steps:
the method comprises the following steps: fish scale cleaning
The collected fish scales are poured into cleaning equipment for cleaning, so that impurities adhered to the surfaces of the fish scales can be cleaned;
step two: pretreatment of
Putting the cleaned fish scales into a reaction kettle, adding an alkali solution into the reaction kettle to enable the pH value of the solution in the reaction kettle to be 7-8, stirring for 1-2 hours to remove mucus on the surfaces of the fish scales, adding a sodium bicarbonate solution to remove lipid substances in the fish scales, and cleaning with deionized water;
step three: extracting by enzymolysis
Adding an enzyme preparation which is 3-8% of the weight of the fed fish scales, carrying out enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, then adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, carrying out enzyme deactivation, and filtering the enzymolysis liquid to obtain primary collagen peptidase hydrolysate;
step four: secondary enzymolysis extraction
Grinding the fish scales filtered out in the third step into powder, adding an enzyme preparation which is 3% of the weight of the added fish scales, performing enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, deactivating enzyme, and filtering the enzymolysis liquid to obtain secondary collagen peptidase hydrolysate;
step five: purifying and concentrating
Stirring and mixing the primary collagen peptidase hydrolyzed solution and the secondary collagen peptidase hydrolyzed solution obtained in the third step and the fourth step, then performing fine filtration by using an ultrafiltration membrane, and performing cooking concentration on the collagen peptide liquid after the fine filtration to obtain a collagen peptide concentrated solution;
step six: drying
And (3) rapidly drying the collagen peptide concentrated solution by utilizing spray drying to obtain a finished collagen peptide powder product.
And in the step one, the cleaning equipment is used for stirring cleaning and aeration cleaning, and the scales can be dispersed during cleaning by utilizing stirring and aeration, so that the phenomenon that the scales are adhered together to influence the cleaning effect is avoided.
And in the second step, the alkali solution is sodium hydroxide or potassium hydroxide solution.
And in the fourth step, the fish scales are grinded into powder by a fish scale grinding machine.
The enzyme preparation in the third step and the fourth step comprises 8 parts of neutral protease, 14 parts of papain, 16 parts of pepsin, 7 parts of trypsin and 5 parts of flavourzyme.
Claims (8)
1. A method for extracting micromolecular collagen peptide by fish scale enzymolysis is characterized by comprising the following steps: the extraction steps of the small molecule collagen peptide are as follows:
the method comprises the following steps: fish scale cleaning
The collected fish scales are poured into cleaning equipment for cleaning, so that impurities adhered to the surfaces of the fish scales can be cleaned;
step two: pretreatment of
Putting the cleaned fish scales into a reaction kettle, adding an alkali solution into the reaction kettle to enable the pH value of the solution in the reaction kettle to be 7-8, stirring for 1-2 hours to remove mucus on the surfaces of the fish scales, adding a sodium bicarbonate solution to remove lipid substances in the fish scales, and cleaning with deionized water;
step three: extracting by enzymolysis
Adding an enzyme preparation which is 3-8% of the weight of the fed fish scales, carrying out enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, then adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, carrying out enzyme deactivation, and filtering the enzymolysis liquid to obtain primary collagen peptidase hydrolysate;
step four: secondary enzymolysis extraction
Grinding the fish scales filtered out in the third step into powder, adding an enzyme preparation which is 3% of the weight of the added fish scales, performing enzymolysis on the pretreated fish scales, adding a sodium bicarbonate solution to adjust the pH value to 5-6, adjusting the temperature to 15-20 ℃, raising the temperature to 85 ℃ after 24 hours of enzymolysis, deactivating enzyme, and filtering the enzymolysis liquid to obtain secondary collagen peptidase hydrolysate;
step five: purifying and concentrating
Stirring and mixing the primary collagen peptidase hydrolyzed solution and the secondary collagen peptidase hydrolyzed solution obtained in the third step and the fourth step, then performing fine filtration by using an ultrafiltration membrane, and performing cooking concentration on the collagen peptide liquid after the fine filtration to obtain a collagen peptide concentrated solution;
step six: drying
And (3) rapidly drying the collagen peptide concentrated solution by utilizing spray drying to obtain a finished collagen peptide powder product.
2. The method for extracting small-molecule collagen peptide by fish scale enzymolysis, as claimed in claim 1, wherein: and the cleaning equipment in the step one is stirring cleaning and aeration cleaning.
3. The method for extracting small-molecule collagen peptide by fish scale enzymolysis, as claimed in claim 1, wherein: and in the second step, the alkali solution is sodium hydroxide or potassium hydroxide solution.
4. The method for extracting small-molecule collagen peptide by fish scale enzymolysis, as claimed in claim 1, wherein: and in the fourth step, the fish scales are grinded into powder by a fish scale grinding machine.
5. The method for extracting small-molecule collagen peptide by fish scale enzymolysis, as claimed in claim 1, wherein: the enzyme preparation in the third step and the fourth step comprises 3-8 parts of neutral protease, 8-14 parts of papain, 10-16 parts of pepsin, 5-7 parts of trypsin and 3-5 parts of flavourzyme.
6. The method for extracting small-molecule collagen peptide by fish scale enzymolysis, as claimed in claim 5, wherein: the enzyme preparation comprises 3 parts of neutral protease, 8 parts of papain, 10 parts of pepsin, 5 parts of trypsin and 3 parts of flavourzyme.
7. The method for extracting small-molecule collagen peptide by fish scale enzymolysis, as claimed in claim 5, wherein: the enzyme preparation comprises 3-8 parts of neutral protease, 11 parts of papain, 13 parts of pepsin, 6 parts of trypsin and 4 parts of flavourzyme.
8. The method for extracting small-molecule collagen peptide by fish scale enzymolysis, as claimed in claim 5, wherein: the enzyme preparation comprises 8 parts of neutral protease, 14 parts of papain, 16 parts of pepsin, 7 parts of trypsin and 5 parts of flavourzyme.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114560927A (en) * | 2022-03-15 | 2022-05-31 | 厦门元之道生物科技有限公司 | Method for producing collagen peptide by subcritical water extraction assisted fish scales and application of method |
| CN117402929A (en) * | 2023-10-27 | 2024-01-16 | 广州市缇梵化妆品有限公司 | Collagen peptide for delaying aging and preparation method and application thereof |
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