CN111944867A - Preparation method of cubilose peptide - Google Patents
Preparation method of cubilose peptide Download PDFInfo
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- CN111944867A CN111944867A CN202010892221.XA CN202010892221A CN111944867A CN 111944867 A CN111944867 A CN 111944867A CN 202010892221 A CN202010892221 A CN 202010892221A CN 111944867 A CN111944867 A CN 111944867A
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title claims abstract description 6
- 102000004190 Enzymes Human genes 0.000 claims abstract description 33
- 108090000790 Enzymes Proteins 0.000 claims abstract description 33
- 229940088598 enzyme Drugs 0.000 claims abstract description 33
- 235000005770 birds nest Nutrition 0.000 claims abstract description 21
- 235000005765 wild carrot Nutrition 0.000 claims abstract description 21
- 239000007788 liquid Substances 0.000 claims abstract description 20
- 238000002156 mixing Methods 0.000 claims abstract description 19
- 244000000626 Daucus carota Species 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 16
- 239000006228 supernatant Substances 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000000047 product Substances 0.000 claims abstract description 12
- 108090000631 Trypsin Proteins 0.000 claims abstract description 9
- 102000004142 Trypsin Human genes 0.000 claims abstract description 9
- 238000001035 drying Methods 0.000 claims abstract description 9
- 238000000926 separation method Methods 0.000 claims abstract description 9
- 229960001322 trypsin Drugs 0.000 claims abstract description 9
- 239000012588 trypsin Substances 0.000 claims abstract description 9
- 108090000317 Chymotrypsin Proteins 0.000 claims abstract description 8
- 102000029816 Collagenase Human genes 0.000 claims abstract description 8
- 108060005980 Collagenase Proteins 0.000 claims abstract description 8
- 229960002376 chymotrypsin Drugs 0.000 claims abstract description 8
- 229960002424 collagenase Drugs 0.000 claims abstract description 8
- 230000009849 deactivation Effects 0.000 claims abstract description 7
- 150000001875 compounds Chemical class 0.000 claims abstract description 6
- 238000010298 pulverizing process Methods 0.000 claims description 11
- 238000009835 boiling Methods 0.000 claims description 7
- 239000008280 blood Substances 0.000 claims description 5
- 210000004369 blood Anatomy 0.000 claims description 5
- 239000000084 colloidal system Substances 0.000 claims description 2
- 238000000265 homogenisation Methods 0.000 claims description 2
- 229920001184 polypeptide Polymers 0.000 abstract description 13
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 13
- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 abstract description 9
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 abstract description 9
- 235000015097 nutrients Nutrition 0.000 abstract description 7
- 210000002268 wool Anatomy 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 238000013341 scale-up Methods 0.000 abstract 1
- 239000000843 powder Substances 0.000 description 19
- 238000005227 gel permeation chromatography Methods 0.000 description 10
- 238000000605 extraction Methods 0.000 description 5
- 230000000415 inactivating effect Effects 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 238000005303 weighing Methods 0.000 description 5
- 210000003746 feather Anatomy 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 210000004209 hair Anatomy 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000004365 Protease Substances 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 239000000413 hydrolysate Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 230000003749 cleanliness Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000019419 proteases Nutrition 0.000 description 2
- 102000011782 Keratins Human genes 0.000 description 1
- 108010076876 Keratins Proteins 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- ZPUCINDJVBIVPJ-LJISPDSOSA-N ***e Chemical compound O([C@H]1C[C@@H]2CC[C@@H](N2C)[C@H]1C(=O)OC)C(=O)C1=CC=CC=C1 ZPUCINDJVBIVPJ-LJISPDSOSA-N 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000035931 haemagglutination Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 238000009966 trimming Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
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
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- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a preparation method of cubilose peptide, which comprises the following steps: s1, pretreating the clean cubilose; s2, mixing the pretreated cubilose and water according to a mass-volume ratio of 1: (4-10), fully mixing, then adjusting the pH to 6-8, adding a complex enzyme accounting for 0.1-1 wt% of the weight of the cubilose, and performing enzymolysis at 25-85 ℃ for 30-180 min to obtain an enzymolysis liquid; s3, carrying out enzyme deactivation on the obtained enzymolysis liquid, and carrying out centrifugal separation to obtain supernatant; and S4, concentrating and drying the obtained supernatant to obtain the cubilose peptide product. The compound enzyme is collagenase, trypsin and chymotrypsin in a mass ratio of (6-12): (2-4): (1-3) mixing. The bird's nest peptide product prepared by the method has the advantages of little loss of nutrient components, more than 60 percent of polypeptide content and more than 30 percent of sialic acid content, no need of manual wool picking, simple operation process, easy control, low cost and easy scale-up production.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a method for preparing cubilose peptide.
Background
The bird's nest, also called as edible bird's nest and edible bird's vegetable, has very high nutritive value and medicinal value, and is called as "oriental caviar". The medicinal effect and medical value of cubilose are also recorded in the herbal standby. Therefore, the cubilose has long been regarded as a health-care food material with homology of medicine and food, and can be used for beautifying, moistening lung and promoting the secretion of saliva or body fluid. Researches show that the cubilose is rich in protein, amino acid, sialic acid, inorganic elements and the like, and has the effects of resisting oxidation, improving immunity, delaying senescence, resisting viruses, inhibiting hemagglutination, regulating intestinal flora and the like.
When the cubilose is processed, an important link is to pick hair, and the input of hair picking cost directly influences the cleanliness and quality of the cubilose. A high-quality swallow cup with up-to-standard cleanliness can be finished only by a series of steps of picking high-quality raw materials → 2-8 ℃ fresh-keeping storage → sorting → trimming corners → removing surface impurities → manually picking coarse hair → cleaning and sterilizing purified water → manually picking fine hair → detecting quality tubes → again finely picking → physical forming → drying by cold wind and the like. Therefore, the artificial wool picking speed is low and the yield is low.
When the edible bird's nest needs to be soaked, stewed and the like, and different soaking and stewing conditions greatly influence the maintenance of the nutrient components of the bird's nest. How to provide a quick and safe method for processing the cubilose with extremely low loss of nutrient components is a problem to be solved urgently in the current cubilose processing industry.
The main chemical components in the edible bird's nest include protein, carbohydrate, sialic acid, amino acid, water, small amount of fat, trace minerals, etc. It is known that the differences in the chemical components and characteristic components of the bird's nest are caused by the differences in the sources, types, picking seasons, and the like of the bird's nest. A great deal of research is carried out on the nutritional ingredients in the cubilose at home and abroad, and the protein and carbohydrate in the cubilose respectively account for 60 percent and 30 percent of the total mass.
Research shows that with the increase of human age, the ability of endocrine protease of human body is gradually reduced, and the digestion and absorption ability of the human body is also reduced, so that the nutrient substances of the edible bird's nest can not be fully utilized by eating the edible bird's nest according to the traditional method.
The bird's nest polypeptide is a protein fragment formed by connecting two or more amino acids through peptide bonds, has the characteristic of easy absorption, and has long been a hot spot for developing health care products and medicines. In the prior art, biological enzymes have been used to hydrolyze edible bird's nest into polypeptides to improve the utilization of nutrients contained in edible bird's nest, such as: the polypeptide is prepared by enzymolysis of cubilose with papain, trypsin and pepsin, and the cubilose is hydrolyzed into polypeptide by protease, which is not only beneficial to absorption of various nutrient substances, but also the polypeptide obtained by enzymolysis can show various biological activities beneficial to human health. However, the existing preparation methods have low polypeptide yield.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a method for preparing cubilose peptide.
A preparation method of cubilose peptide comprises the following steps:
s1, pretreating the clean cubilose;
s2, mixing the pretreated cubilose and water according to a mass-volume ratio of 1: (4-10), fully mixing, then adjusting the pH to 6-8, adding a complex enzyme accounting for 0.1-1 wt% of the weight of the cubilose, and performing enzymolysis at 25-85 ℃ for 30-180 min to obtain an enzymolysis liquid;
s3, carrying out enzyme deactivation on the obtained enzymolysis liquid, and carrying out centrifugal separation to obtain supernatant;
s4, concentrating and drying the obtained supernatant to obtain a bird' S nest peptide product;
the compound enzyme is collagenase, trypsin and chymotrypsin in a mass ratio of (6-12): (2-4): (1-3) mixing.
Preferably, in step S1, the pretreatment is a pulverization treatment, and the pulverization treatment is one or a combination of two or more of ultrafine pulverization, colloid mill pulverization, wall-breaking pulverization, ball mill pulverization, and homogenizer homogenization.
Preferably, the clean cubilose is crushed to 80-150 meshes.
Preferably, in the step S2, the adding amount of the complex enzyme is 0.2-0.8 wt% of the weight of the cubilose, the enzymolysis temperature is 30-80 ℃, and the enzymolysis time is 60-150 min.
Preferably, in step S3, the enzyme deactivation is performed by boiling the enzymatic hydrolysate at a high temperature or adjusting the pH of the enzymatic hydrolysate to 12 to 14.
Preferably, in step S3, the enzyme deactivation is performed by boiling the enzymatic hydrolysate in a boiling water bath for 10-20 min.
Preferably, the cubilose is one of white cubilose, blood cubilose or hairy cubilose.
Compared with the prior art, the invention has the beneficial effects that:
(1) the method has strong universality, the types of the cubilose are not limited, the white cubilose, the blood cubilose and the hairy cubilose can all obtain expected cubilose peptide products, when the hairy cubilose is used as a cubilose raw material, the feathers still exist in a solid form in the step S2 because the used compound enzyme can not carry out enzymolysis on the keratin of the feathers, and the feathers can be completely removed through solid-liquid separation in the step S3, so that manual feather picking is not needed;
(2) the bird's nest peptide product prepared by the method has little loss of nutrient components, the polypeptide content is more than 60 percent, and the sialic acid content is more than 30 percent;
(3) the complex enzymes used in the invention all belong to food additives allowed by the nation, and can be used in the food industry without treatment;
(4) the method of the invention does not need manual wool picking, has simple operation process, easy control, standardized equipment, low cost, easy enlarged production and convenient popularization and application.
Detailed Description
The invention is illustrated below with reference to specific examples. It will be understood by those skilled in the art that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention in any way.
Example 1
S1, weighing 500g of clean collophanite, and crushing the collophanite to 120 meshes to obtain edible bird nest powder;
s2, adding 5kg of water into the obtained cubilose powder, then placing the cubilose powder into an extraction tank, stirring and mixing the mixture at 50 ℃, then adjusting the pH to 8, then adding 5g of compound enzyme, and carrying out enzymolysis for 120min to obtain an enzymolysis liquid, wherein the compound enzyme is collagenase, trypsin and chymotrypsin according to a mass ratio of 8: 2: 3, mixing to obtain the mixture;
s3, placing the enzymolysis liquid obtained in the step S2 in a boiling water bath to boil for 10min, inactivating enzyme, performing centrifugal separation, and collecting supernatant;
s4, concentrating and drying the supernatant collected in the step S3 to obtain 471g of cubilose peptide product, and detecting by GPC gel permeation chromatography, wherein the content of the polypeptide is 62.1%, and the content of the sialic acid is 31.1%.
Example 2
S1, weighing 500g of clean collophanite, and crushing the collophanite to 150 meshes to obtain edible bird nest powder;
s2, adding 2kg of water into the obtained cubilose powder, then placing the cubilose powder into an extraction tank, stirring and mixing the cubilose powder at the temperature of 80 ℃, then adjusting the pH value to 6, then adding 0.5g of complex enzyme, and carrying out enzymolysis for 120min to obtain an enzymolysis liquid, wherein the complex enzyme is collagenase, trypsin and chymotrypsin according to the mass ratio of 10: 4: 1, mixing the raw materials;
s3, placing the enzymolysis liquid obtained in the step S2 in a boiling water bath to boil for 10min, inactivating enzyme, performing centrifugal separation, and collecting supernatant;
s4, concentrating and drying the supernatant collected in the step S3 to obtain 467g of cubilose peptide product, and detecting by GPC gel permeation chromatography, wherein the content of the polypeptide is 63.2 percent and the content of the sialic acid is 31.8 percent.
Example 3
S1, weighing 500g of clean collophanite, and crushing the collophanite to 80 meshes to obtain edible bird nest powder;
s2, adding 3kg of water into the obtained cubilose powder, then placing the cubilose powder into an extraction tank, stirring and mixing the cubilose powder at 25 ℃, then adjusting the pH value to 7, then adding 2.5g of complex enzyme, and carrying out enzymolysis for 180min to obtain an enzymolysis liquid, wherein the complex enzyme is collagenase, trypsin and chymotrypsin according to a mass ratio of 8: 4: 3, mixing to obtain the mixture;
s3, placing the enzymolysis liquid obtained in the step S2 in a boiling water bath to boil for 15min, inactivating enzyme, performing centrifugal separation, and collecting supernatant;
s4, concentrating and drying the supernatant collected in the step S3 to obtain 465g of cubilose peptide product, and detecting by GPC gel permeation chromatography, wherein the content of the polypeptide is 62.5 percent, and the content of the sialic acid is 30.6 percent.
Example 4
S1, weighing 500g of clean blood swallow, and crushing the blood swallow to 100 meshes to obtain cubilose powder;
s2, adding 2kg of water into the obtained cubilose powder, then placing the cubilose powder into an extraction tank, stirring and mixing the cubilose powder at 45 ℃, adding 4.0g of complex enzyme, and carrying out enzymolysis for 150min to obtain an enzymolysis liquid, wherein the complex enzyme is collagenase, trypsin and chymotrypsin according to a mass ratio of 6: 2: 1, mixing the raw materials;
s3, adjusting the pH value of the enzymolysis liquid obtained in the step S2 to 12, inactivating enzyme, carrying out centrifugal separation, and collecting supernatant;
s4, concentrating and drying the supernatant collected in the step S3 to obtain 466g of cubilose peptide product, and detecting by GPC gel permeation chromatography, wherein the content of the polypeptide is 62.7 percent, and the content of the sialic acid is 31.7 percent.
Example 5
S1, weighing 500g of clean white swallow, and crushing the white swallow to 120 meshes to obtain cubilose powder;
s2, adding 4kg of water into the obtained cubilose powder, then placing the cubilose powder into an extraction tank, stirring and mixing the cubilose powder at 85 ℃, then adjusting the pH value to 7, then adding 1.0g of complex enzyme, and carrying out enzymolysis for 60min to obtain an enzymolysis liquid, wherein the complex enzyme is collagenase, trypsin and chymotrypsin according to the mass ratio of 12: 2: 3, mixing to obtain the mixture;
s3, adjusting the pH value of the enzymolysis liquid obtained in the step S2 to 13, inactivating enzyme, carrying out centrifugal separation, and collecting supernatant;
s4, concentrating and drying the supernatant collected in the step S3 to obtain 467g of cubilose peptide product, and detecting by GPC gel permeation chromatography, wherein the content of the polypeptide is 61.6 percent and the content of the sialic acid is 31.4 percent.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (7)
1. The preparation method of the cubilose peptide is characterized by comprising the following steps:
s1, pretreating the clean cubilose;
s2, mixing the pretreated cubilose and water according to a mass-volume ratio of 1: (4-10), fully mixing, then adjusting the pH to 6-8, adding a complex enzyme accounting for 0.1-1 wt% of the weight of the cubilose, and performing enzymolysis at 25-85 ℃ for 30-180 min to obtain an enzymolysis liquid;
s3, carrying out enzyme deactivation on the obtained enzymolysis liquid, and carrying out centrifugal separation to obtain supernatant;
s4, concentrating and drying the obtained supernatant to obtain a bird' S nest peptide product;
the compound enzyme is collagenase, trypsin and chymotrypsin in a mass ratio of (6-12): (2-4): (1-3) mixing.
2. The method of claim 1, wherein the pretreatment is pulverization in step S1, and the pulverization is performed by one or a combination of two or more of micronization, colloid mill pulverization, wall-breaking pulverization, ball mill pulverization, and homogenizer homogenization.
3. The method for preparing bird's nest peptide according to claim 2, characterized in that clean bird's nests are crushed to 80-150 meshes.
4. The method for preparing cubilose peptide according to claim 1, wherein in step S2, the adding amount of the complex enzyme is 0.2-0.8 wt% of the weight of cubilose, the enzymolysis temperature is 30-80 ℃, and the enzymolysis time is 60-150 min.
5. The method for preparing cubilose peptide according to claim 1, wherein in step S3, the enzyme deactivation mode is to boil the enzymolysis liquid at high temperature or to adjust the pH of the enzymolysis liquid to 12-14.
6. The method for preparing cubilose peptide according to claim 5, wherein in step S3, the enzyme deactivation mode is to boil the enzymolysis liquid in a boiling water bath for 10-20 min.
7. The method for preparing cubilose peptide according to claim 1, wherein the cubilose is one of white cubilose, blood cubilose or hairy cubilose.
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| CN202010892221.XA CN111944867A (en) | 2020-08-31 | 2020-08-31 | Preparation method of cubilose peptide |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113243526A (en) * | 2021-05-06 | 2021-08-13 | 海南大洲金丝燕产业集团有限公司 | Edible bird's nest acid extraction method for edible bird's nest processing |
| CN113322297A (en) * | 2021-05-31 | 2021-08-31 | 菏泽牡丹产业技术研究院 | Preparation method of peony and bird's nest composite protein peptide |
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Cited By (2)
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| CN113243526A (en) * | 2021-05-06 | 2021-08-13 | 海南大洲金丝燕产业集团有限公司 | Edible bird's nest acid extraction method for edible bird's nest processing |
| CN113322297A (en) * | 2021-05-31 | 2021-08-31 | 菏泽牡丹产业技术研究院 | Preparation method of peony and bird's nest composite protein peptide |
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