CN107397954A - The purposes for the function small peptide extracted from deep-sea fish - Google Patents
The purposes for the function small peptide extracted from deep-sea fish Download PDFInfo
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- CN107397954A CN107397954A CN201710640809.4A CN201710640809A CN107397954A CN 107397954 A CN107397954 A CN 107397954A CN 201710640809 A CN201710640809 A CN 201710640809A CN 107397954 A CN107397954 A CN 107397954A
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 85
- 241000251468 Actinopterygii Species 0.000 title claims abstract description 33
- 108091005804 Peptidases Proteins 0.000 claims abstract description 46
- 239000004365 Protease Substances 0.000 claims abstract description 43
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 43
- 108010035532 Collagen Proteins 0.000 claims abstract description 33
- 102000008186 Collagen Human genes 0.000 claims abstract description 33
- 229920001436 collagen Polymers 0.000 claims abstract description 33
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 27
- 241000206613 Pyropia yezoensis Species 0.000 claims abstract description 25
- 102000004190 Enzymes Human genes 0.000 claims abstract description 23
- 108090000790 Enzymes Proteins 0.000 claims abstract description 23
- 238000001728 nano-filtration Methods 0.000 claims abstract description 18
- 239000000796 flavoring agent Substances 0.000 claims abstract description 16
- 235000019634 flavors Nutrition 0.000 claims abstract description 16
- 238000004108 freeze drying Methods 0.000 claims abstract description 11
- 238000001556 precipitation Methods 0.000 claims abstract description 10
- 235000013305 food Nutrition 0.000 claims abstract description 9
- 239000002537 cosmetic Substances 0.000 claims abstract description 7
- 239000003814 drug Substances 0.000 claims abstract description 7
- 230000036541 health Effects 0.000 claims abstract description 7
- 239000003223 protective agent Substances 0.000 claims abstract description 7
- 239000000047 product Substances 0.000 claims description 16
- 230000000975 bioactive effect Effects 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 230000009849 deactivation Effects 0.000 claims description 10
- 239000000706 filtrate Substances 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 230000014759 maintenance of location Effects 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- IZXIZTKNFFYFOF-UHFFFAOYSA-N 2-Oxazolidone Chemical class O=C1NCCO1 IZXIZTKNFFYFOF-UHFFFAOYSA-N 0.000 claims description 9
- 239000004698 Polyethylene Substances 0.000 claims description 9
- KJADKKWYZYXHBB-XBWDGYHZSA-N Topiramic acid Chemical compound C1O[C@@]2(COS(N)(=O)=O)OC(C)(C)O[C@H]2[C@@H]2OC(C)(C)O[C@@H]21 KJADKKWYZYXHBB-XBWDGYHZSA-N 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 9
- -1 polyethylene Polymers 0.000 claims description 9
- 229920000573 polyethylene Polymers 0.000 claims description 9
- 229960004394 topiramate Drugs 0.000 claims description 9
- 230000007062 hydrolysis Effects 0.000 claims description 8
- 238000006460 hydrolysis reaction Methods 0.000 claims description 8
- 238000010612 desalination reaction Methods 0.000 claims description 7
- 238000007710 freezing Methods 0.000 claims description 7
- 230000008014 freezing Effects 0.000 claims description 7
- 239000000654 additive Substances 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 5
- 239000002244 precipitate Substances 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 230000005611 electricity Effects 0.000 claims 1
- 235000019419 proteases Nutrition 0.000 abstract description 38
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 abstract description 22
- 210000004051 gastric juice Anatomy 0.000 abstract description 11
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 10
- 239000001257 hydrogen Substances 0.000 abstract description 10
- 241001465754 Metazoa Species 0.000 abstract description 8
- 238000010521 absorption reaction Methods 0.000 abstract description 6
- 210000000845 cartilage Anatomy 0.000 abstract description 6
- 210000002808 connective tissue Anatomy 0.000 abstract description 6
- 239000003623 enhancer Substances 0.000 abstract description 6
- 230000002500 effect on skin Effects 0.000 abstract description 5
- 102000035195 Peptidases Human genes 0.000 abstract description 3
- 235000019833 protease Nutrition 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 description 15
- 108090000623 proteins and genes Proteins 0.000 description 15
- 150000001413 amino acids Chemical class 0.000 description 13
- 210000003491 skin Anatomy 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 239000012528 membrane Substances 0.000 description 6
- 230000010534 mechanism of action Effects 0.000 description 5
- 230000007071 enzymatic hydrolysis Effects 0.000 description 4
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 238000005411 Van der Waals force Methods 0.000 description 3
- 210000000988 bone and bone Anatomy 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 235000013601 eggs Nutrition 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
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- 238000011105 stabilization Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- YBPLKDWJFYCZSV-ZLUOBGJFSA-N Ala-Asn-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CS)C(=O)O)N YBPLKDWJFYCZSV-ZLUOBGJFSA-N 0.000 description 1
- UQJUGHFKNKGHFQ-VZFHVOOUSA-N Ala-Cys-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UQJUGHFKNKGHFQ-VZFHVOOUSA-N 0.000 description 1
- MQQLYEHXSBJTRK-FXQIFTODSA-N Cys-Val-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CS)N MQQLYEHXSBJTRK-FXQIFTODSA-N 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- HBGKOLSGLYMWSW-DCAQKATOSA-N His-Pro-Cys Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CN=CN2)N)C(=O)N[C@@H](CS)C(=O)O HBGKOLSGLYMWSW-DCAQKATOSA-N 0.000 description 1
- DGLAHESNTJWGDO-SRVKXCTJSA-N His-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N DGLAHESNTJWGDO-SRVKXCTJSA-N 0.000 description 1
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- XUDRHBPSPAPDJP-SRVKXCTJSA-N Ser-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CO XUDRHBPSPAPDJP-SRVKXCTJSA-N 0.000 description 1
- OSFZCEQJLWCIBG-BZSNNMDCSA-N Ser-Tyr-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OSFZCEQJLWCIBG-BZSNNMDCSA-N 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 102000034240 fibrous proteins Human genes 0.000 description 1
- 108091005899 fibrous proteins Proteins 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 108010040030 histidinoalanine Proteins 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 235000002020 sage Nutrition 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 1
- 230000002087 whitening effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/461—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from fish
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention discloses a kind of purposes for the function small peptide extracted from deep-sea fish, as the protective agent or absorption enhancers of collagen, for cosmetics or health products or food or feed or medicine.The purposes for the function small peptide extracted from deep-sea fish, including stepwise discretization, isoelectric precipitation, second ultrafiltration and nanofiltration and freeze-drying.Digested using marine protein enzyme and land proteinase synergy, first using Porphyra yezoensis protease, continue to digest using flavor protease on this basis, can obtain the function small peptide that collagen can be protected not decomposed by hydrochloric acid in gastric juice.Have the beneficial effect that：Connected between obtained function small peptide and collagen by intermolecular force and hydrogen bond; function small peptide is in collagen molecules surface attachment; wrapped up, collagen can be protected not decomposed by hydrochloric acid in gastric juice, and it can be promoted to reach animal connective tissue, cartilage and dermal layer of the skin.
Description
Technical field
The present invention relates to small peptide preparation field, specifically a kind of purposes for the function small peptide extracted from deep-sea fish.
Background technology
Naturally occurring Marine Bioactive Peptides due in organism content it is low, and extract difficult, it is difficult to realize a large amount of
Needed for production supply, chemical artificial synthetic polypeptide is costly.So sight is more invested exploitation proteolysis production by people
Come in this approach of thing acquisition active peptides.Because the environment and terrestrial life of marine organisms existence are entirely different, such as high pressure, low
The extreme environment such as temperature, high temperature and high salt, in order to adapt to these extreme marine biological environments, marine protein matter no matter amino
The composition or the sequence of amino acid of acid are all very different with terrestrial life albumen, meanwhile, marine protein resource without
By being all far longer than land protein resource in type and quantity, and do not developed well.Miscellaneous ocean egg
In casamino acid sequence, potential many amino acid sequences with bioactivity, with special protease hydrolytic, just discharge
Active peptide fragment.Marine protein resource is the important of the important albumen group food of the 21 century mankind and bioactive substance
Source.The current marine protein total resources in China comes out at the top in countries in the world, but the protide aquatic products that China is current
In product, in addition to a part is directly edible, major part is made food by simple process technology and enters market, and process technology falls
Afterwards, product structure is single, and value-added content of product is low so that the competitiveness of product in the international market is poor.Therefore, China is badly in need of
Marine protein resource is optimized and utilized and high-valued processing.
Collagen is also known as collagen, is the fibrous proteins being mutually wound by three peptide chains, is a kind of glycoprotein,
Containing substantial amounts of glycine, proline, hydroxyproline and sugar, the tissue such as skin, bone, cartilage and tendon of animal is widely present in
In, it is the most class protein of body content.Collagen can be used for repairing the sides such as skin scar, anti-ageing, light spot and whitening
Face, it is beauty sage's product, but by edible way replenishing collagen, its utilization rate is extremely low, can largely be decomposed by body, loses
Deactivation.
The content of the invention
It is an object of the invention to provide a kind of purposes for the function small peptide extracted from deep-sea fish, as collagen
Protective agent or absorption enhancers, for cosmetics or health products or food or feed or medicine.Extracting obtained function small peptide can protect
Shield collagen is not decomposed by hydrochloric acid in gastric juice, and it can be promoted to reach animal connective tissue, cartilage and dermal layer of the skin.
The present invention is directed to the problem of being mentioned in background technology, and the technical scheme taken is：The function of being extracted from deep-sea fish
The purposes of small peptide, as the protective agent or absorption enhancers of collagen, for cosmetics or health products or food or feed or medicine
Product.
The purposes for the function small peptide extracted from deep-sea fish, including stepwise discretization, isoelectric precipitation, second ultrafiltration and nanofiltration
And freeze-drying.Stepwise discretization step is that pure water homogenate is added in deep-sea fish minced fillet, adds Porphyra yezoensis protease and work
Property small peptide enzymolysis, adjust pH after add flavor protease enzymolysis, obtain enzymolysis liquid.Marine protein matter no matter the group of amino acid
Sequence into still amino acid is all very different with terrestrial life albumen, can release with marine protein enzyme enzymolysis marine product
Many different active peptide segments.Porphyra yezoensis protease is first used, continues to digest using flavor protease on this basis, can obtain
To the function small peptide that collagen can be protected not decomposed by hydrochloric acid in gastric juice.Do not have at present and obtain above-mentioned function using other protease hydrolyzeds
The report of small peptide.Digested, had significant creative using marine protein enzyme and land proteinase synergy.
20 ~ 30 times of volume of water are added in deep-sea fish minced fillet, the addition of Porphyra yezoensis protease is 5 ~ 8wt ‰, enzymolysis temperature
25 ~ 35 DEG C of degree, pH are 7.0 ~ 9.0, and enzymolysis time is 30 ~ 40min, and enzymolysis terminates rear enzyme deactivation.Under above-mentioned enzymatic hydrolysis condition, streak is purple
The enzymolysis efficiency of dish protease is high will not to cause the wasting of resources again, for most preferably condition.
Flavor protease addition is 1 ~ 3wt ﹪, and enzymolysis pH is 6.0 ~ 7.0, and hydrolysis temperature is 52 ~ 60 DEG C, enzymolysis time
For 15 ~ 25min, enzymolysis terminates rear enzyme deactivation, filters to take filtrate.Under above-mentioned enzymatic hydrolysis condition, the enzymolysis efficiency height of flavor protease is again
The wasting of resources will not be caused, for most preferably condition.
The addition of bioactive peptide is 10 ~ 20 ﹪ of Porphyra yezoensis protease quality, and amino acid sequence is
HSHACTSYYHPCSKLLCVCANCSR.Above-mentioned bioactive peptide can change Porphyra yezoensis egg by Van der Waals force and electrostatic interaction
The tertiary structure of white enzyme, makes the enzymolysis vigor of Porphyra yezoensis protease be improved significantly.
For isoelectric precipitation step for enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.5 ~ 4.8, and centrifuging and taking is sunk after standing
Form sediment.Under above-mentioned pH environment, function small peptide is in electroneutral, is slightly soluble in water, most of to be isolated with precipitation form from the aqueous solution
Come.
Second ultrafiltration and nano-filtration step are first to use 30 ~ 40kD of retention relative molecular mass film ultrafiltration, then using retention
4 ~ 8kD of relative molecular mass film ultrafiltration, filtered solution is finally subjected to nanofiltration desalination.In order to prevent Pore Blocking or membrane damage,
Ultrafiltration is first carried out using big porous ultrafiltration membrane, then using small porous ultrafiltration membrane ultrafiltration, effectively removes the impurity that molecular weight is more than 4 ~ 8kD.
Nanofiltration can be more thoroughly remove solution in inorganic salts and molecular weight be less than 200Da impurity, function small peptide is further carried out
Purifying.
It is freeze-dried in step and adds 20 ~ 35wt% polyethylene Topiramate Los oxazolidinone as lyophilisation additives, is freeze-dried and obtains work(
Can small peptide freeze-dried powder.Function small peptide need containing suitable residual moisture come be maintained at storage during property stabilization, excessively
Drying by the hydrogen bond for making protein molecule surface and polar group exposure and be denatured.In freezing dry process, polyethylene is added
Topiramate Los oxazolidinone, can the albumen stretching, extension of defencive function small peptide, quaternary structure, so as to being protected to the small peptide activity of function.Above-mentioned work(
It can be connected between small peptide and collagen by intermolecular force and hydrogen bond, function small peptide is attached on collagen molecules surface
, wrapped up, collagen can be protected not decomposed by hydrochloric acid in gastric juice, and it can be promoted to reach animal connective tissue, cartilage and skin
Skin corium, the specific mechanism of action are still not clear.
Compared with prior art, the advantage of the invention is that：
1. the composition of marine protein matter amino acid or the sequence of amino acid all have very big with terrestrial life albumen
Difference, many different active peptide segments are can release with marine protein enzyme enzymolysis marine product.Porphyra yezoensis protease is first used,
Continue using dividing flavor protease to digest, to can obtain the function small peptide that collagen can be protected not decomposed by hydrochloric acid in gastric juice on the basis of this.
Do not have at present and obtain the report of above-mentioned function small peptide using other protease hydrolyzeds.Assisted using marine protein enzyme and land protease
With enzymolysis, have significant creative.
2. amino acid sequence, which is HSHACTSYYHPCSKLLCVCANCSR bioactive peptide, can pass through Van der Waals force and electrostatic
Act on to change the tertiary structure of Porphyra yezoensis protease, the enzymolysis vigor of Porphyra yezoensis protease is improved significantly.
3. being connected between above-mentioned function small peptide and collagen by intermolecular force and hydrogen bond, function small peptide is in glue
Former protein molecular surface attachment, is wrapped up, and collagen can be protected not decomposed by hydrochloric acid in gastric juice, and it can be promoted to reach animal knot
Tissue, cartilage and dermal layer of the skin are formed, the specific mechanism of action is still not clear.
Embodiment
The present invention program is described further below by embodiment：
Embodiment 1：
The purposes for the function small peptide extracted from deep-sea fish, as the protective agent or absorption enhancers of collagen, for cosmetics
Or health products or food or feed or medicine.
The purposes for the function small peptide extracted from deep-sea fish, including stepwise discretization, isoelectric precipitation, second ultrafiltration and nanofiltration
And freeze-drying.Stepwise discretization step is that pure water homogenate is added in deep-sea fish minced fillet, adds Porphyra yezoensis protease and work
Property small peptide enzymolysis, adjust pH after add flavor protease enzymolysis, obtain enzymolysis liquid.Marine protein matter no matter the group of amino acid
Sequence into still amino acid is all very different with terrestrial life albumen, can release with marine protein enzyme enzymolysis marine product
Many different active peptide segments.Porphyra yezoensis protease is first used, continues to digest using flavor protease on this basis, can obtain
To the function small peptide that collagen can be protected not decomposed by hydrochloric acid in gastric juice.Do not have at present and obtain above-mentioned function using other protease hydrolyzeds
The report of small peptide.Digested, had significant creative using marine protein enzyme and land proteinase synergy.
20 ~ 30 times of volume of water are added in deep-sea fish minced fillet, the addition of Porphyra yezoensis protease is 5 ~ 8wt ‰, enzymolysis temperature
25 ~ 35 DEG C of degree, pH are 7.0 ~ 9.0, and enzymolysis time is 30 ~ 40min, and enzymolysis terminates rear enzyme deactivation.Under above-mentioned enzymatic hydrolysis condition, streak is purple
The enzymolysis efficiency of dish protease is high will not to cause the wasting of resources again, for most preferably condition.
Flavor protease addition is 1 ~ 3wt ﹪, and enzymolysis pH is 6.0 ~ 7.0, and hydrolysis temperature is 52 ~ 60 DEG C, enzymolysis time
For 15 ~ 25min, enzymolysis terminates rear enzyme deactivation, filters to take filtrate.Under above-mentioned enzymatic hydrolysis condition, the enzymolysis efficiency height of flavor protease is again
The wasting of resources will not be caused, for most preferably condition.
The addition of bioactive peptide is 10 ~ 20 ﹪ of Porphyra yezoensis protease quality, and amino acid sequence is
HSHACTSYYHPCSKLLCVCANCSR.Above-mentioned bioactive peptide can change Porphyra yezoensis egg by Van der Waals force and electrostatic interaction
The tertiary structure of white enzyme, makes the enzymolysis vigor of Porphyra yezoensis protease be improved significantly.
For isoelectric precipitation step for enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.5 ~ 4.8, and centrifuging and taking is sunk after standing
Form sediment.Under above-mentioned pH environment, function small peptide is in electroneutral, is slightly soluble in water, most of to be isolated with precipitation form from the aqueous solution
Come.
Second ultrafiltration and nano-filtration step are first to use 30 ~ 40kD of retention relative molecular mass film ultrafiltration, then using retention
4 ~ 8kD of relative molecular mass film ultrafiltration, filtered solution is finally subjected to nanofiltration desalination.In order to prevent Pore Blocking or membrane damage,
Ultrafiltration is first carried out using big porous ultrafiltration membrane, then using small porous ultrafiltration membrane ultrafiltration, effectively removes the impurity that molecular weight is more than 4 ~ 8kD.
Nanofiltration can be more thoroughly remove solution in inorganic salts and molecular weight be less than 200Da impurity, function small peptide is further carried out
Purifying.
It is freeze-dried in step and adds 20 ~ 35wt% polyethylene Topiramate Los oxazolidinone as lyophilisation additives, is freeze-dried and obtains work(
Can small peptide freeze-dried powder.Function small peptide need containing suitable residual moisture come be maintained at storage during property stabilization, excessively
Drying by the hydrogen bond for making protein molecule surface and polar group exposure and be denatured.In freezing dry process, polyethylene is added
Topiramate Los oxazolidinone, can the albumen stretching, extension of defencive function small peptide, quaternary structure, so as to being protected to the small peptide activity of function.Above-mentioned work(
It can be connected between small peptide and collagen by intermolecular force and hydrogen bond, function small peptide is attached on collagen molecules surface
, wrapped up, collagen can be protected not decomposed by hydrochloric acid in gastric juice, and it can be promoted to reach animal connective tissue, cartilage and skin
Skin corium, the specific mechanism of action are still not clear.
Embodiment 2：
The purposes for the function small peptide extracted from deep-sea fish, as the protective agent or absorption enhancers of collagen, for cosmetics
Or health products or food or feed or medicine.
The most preferred method of abstraction function small peptide, comprises the following steps from deep-sea fish：
1）Stepwise discretization：Pure water homogenate is added in deep-sea fish minced fillet, adds Porphyra yezoensis protease and bioactive peptide enzymolysis,
Flavor protease enzymolysis is added after adjusting pH, obtains enzymolysis liquid.26 times of volume of water, Porphyra yezoensis albumen are added in deep-sea fish minced fillet
The addition of enzyme is 6wt ‰, 32 DEG C, pH 8.0, enzymolysis time 34min of hydrolysis temperature, digests and terminates rear enzyme deactivation.Flavor egg
White enzyme addition is 2wt ﹪, and enzymolysis pH is 6.4, and hydrolysis temperature is 55 DEG C, enzymolysis time 21min, and enzymolysis terminates rear enzyme deactivation,
Filter to take filtrate.The addition of bioactive peptide is 16 ﹪ of Porphyra yezoensis protease quality, and amino acid sequence is
HSHACTSYYHPCSKLLCVCANCSR；
2）Isoelectric precipitation：Enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.7, and centrifuging and taking precipitates after standing；
3）Second ultrafiltration and nanofiltration：First using retention relative molecular mass 35kD film ultrafiltration, then using retention average molecular matter
6kD film ultrafiltration is measured, filtered solution is finally subjected to nanofiltration desalination；
4）Freeze-drying：30wt% polyethylene Topiramate Los oxazolidinone is added in filtered solution after nanofiltration desalination as lyophilisation additives, freezing
It is dried to obtain function small peptide freeze-dried powder.Function small peptide needs to be maintained at property during storage containing suitable residual moisture
Stable, the hydrogen bond for making protein molecule surface and polar group exposure are denatured by excessive drying.In freezing dry process, add
Enter polyethylene Topiramate Los oxazolidinone, can the albumen stretching, extension of defencive function small peptide, quaternary structure, so as to being protected to the small peptide activity of function
Shield.Connected between above-mentioned function small peptide and collagen by intermolecular force and hydrogen bond, function small peptide is in collagen point
Sub- surface attachment, is wrapped up, and collagen can be protected not decomposed by hydrochloric acid in gastric juice, and it can be promoted to reach animal connective tissue, soft
Bone and dermal layer of the skin, the specific mechanism of action are still not clear.
Embodiment 3：
The purposes for the function small peptide extracted from deep-sea fish, as the protective agent or absorption enhancers of collagen, for cosmetics
Or health products or food or feed or medicine.
The most preferred method of abstraction function small peptide, comprises the following steps from deep-sea fish：
1）Stepwise discretization：Pure water homogenate is added in deep-sea fish minced fillet, adds Porphyra yezoensis protease and bioactive peptide enzymolysis,
Flavor protease enzymolysis is added after adjusting pH, obtains enzymolysis liquid.30 times of volume of water, Porphyra yezoensis albumen are added in deep-sea fish minced fillet
The addition of enzyme is 5.5wt ‰, 32 DEG C, pH 8.0, enzymolysis time 34min of hydrolysis temperature, digests and terminates rear enzyme deactivation.Flavor
Protease addition is 1.8wt ﹪, and enzymolysis pH is 6.4, and hydrolysis temperature is 55 DEG C, enzymolysis time 21min, and enzymolysis goes out after terminating
Enzyme, filter to take filtrate.The addition of bioactive peptide is 16 ﹪ of Porphyra yezoensis protease quality, and amino acid sequence is
HSHACTSYYHPCSKLLCVCANCSR；
2）Isoelectric precipitation：Enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.7, and centrifuging and taking precipitates after standing；
3）Second ultrafiltration and nanofiltration：First using retention relative molecular mass 35kD film ultrafiltration, then using retention average molecular matter
6kD film ultrafiltration is measured, filtered solution is finally subjected to nanofiltration desalination；
4）Freeze-drying：30wt% polyethylene Topiramate Los oxazolidinone is added in filtered solution after nanofiltration desalination as lyophilisation additives, freezing
It is dried to obtain function small peptide freeze-dried powder.Function small peptide needs to be maintained at property during storage containing suitable residual moisture
Stable, the hydrogen bond for making protein molecule surface and polar group exposure are denatured by excessive drying.In freezing dry process, add
Enter polyethylene Topiramate Los oxazolidinone, can the albumen stretching, extension of defencive function small peptide, quaternary structure, so as to being protected to the small peptide activity of function
Shield.Connected between above-mentioned function small peptide and collagen by intermolecular force and hydrogen bond, function small peptide is in collagen point
Sub- surface attachment, is wrapped up, and collagen can be protected not decomposed by hydrochloric acid in gastric juice, and it can be promoted to reach animal connective tissue, soft
Bone and dermal layer of the skin, the specific mechanism of action are still not clear.
Routine operation in the operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme is described in detail embodiment described above, it should be understood that it is described above only
For the specific embodiment of the present invention, it is not intended to limit the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should be included in the scope of the protection.
SEQUENCE LISTING
<110>Lanxi City silence bio tech ltd
<120>The purposes for the function small peptide extracted from deep-sea fish
<130> 1
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 24
<212> PRT
<213>It is artificial synthesized
<400> 1
His Ser His Ala Cys Thr Ser Tyr Tyr His Pro Cys Ser Lys Leu Leu
1 5 10 15
Cys Val Cys Ala Asn Cys Ser Arg
20
Claims (8)
1. the purposes for the function small peptide extracted from deep-sea fish, it is characterised in that：Protective agent or rush as collagen absorb
Agent, for cosmetics or health products or food or feed or medicine.
2. the purposes of the function small peptide according to claim 1 extracted from deep-sea fish, it is characterised in that：Described function
The preparation method of small peptide, including stepwise discretization, isoelectric precipitation, second ultrafiltration and nanofiltration and freeze-drying；
Wherein, described stepwise discretization step is that pure water homogenate is added in deep-sea fish minced fillet, adds Porphyra yezoensis protease
Digested with bioactive peptide, add flavor protease enzymolysis after adjusting pH, obtain enzymolysis liquid.
3. the method for the function small peptide according to claim 2 extracted from deep-sea fish, it is characterised in that：Described deep-sea
Add 20 ~ 30 times of volume of water in fish minced fillet, the addition of Porphyra yezoensis protease is 5 ~ 8wt ‰, 25 ~ 35 DEG C of hydrolysis temperature, pH
For 7.0 ~ 9.0, enzymolysis time is 30 ~ 40min, and enzymolysis terminates rear enzyme deactivation.
4. the method for the function small peptide according to claim 2 extracted from deep-sea fish, it is characterised in that：Described flavor
Protease addition is 1 ~ 3wt ﹪, and enzymolysis pH is 6.0 ~ 7.0, and hydrolysis temperature is 52 ~ 60 DEG C, and enzymolysis time is 15 ~ 25min, enzyme
Solution terminates rear enzyme deactivation, filters to take filtrate.
5. the method for the function small peptide according to claim 2 extracted from deep-sea fish, it is characterised in that：Described activity
The addition of small peptide is 10 ~ 20 ﹪ of Porphyra yezoensis protease quality, and amino acid sequence is
HSHACTSYYHPCSKLLCVCANCSR。
6. the method for the function small peptide according to claim 2 extracted from deep-sea fish, it is characterised in that：Described waits electricity
For point settling step for enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.5 ~ 4.8, and centrifuging and taking precipitates after standing.
7. the method for the function small peptide according to claim 2 extracted from deep-sea fish, it is characterised in that：Described is secondary
Ultrafiltration and nano-filtration step are first to use 30 ~ 40kD of retention relative molecular mass film ultrafiltration, then using retention relative molecular mass 4
~ 8kD film ultrafiltration, filtered solution is finally subjected to nanofiltration desalination.
8. the method for the function small peptide according to claim 2 extracted from deep-sea fish, it is characterised in that：Described freezing
20 ~ 35wt% polyethylene Topiramate Los oxazolidinones are added in drying steps as lyophilisation additives.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108077939A (en) * | 2017-11-29 | 2018-05-29 | 兰溪市沉默生物科技有限公司 | The deep-sea fish flesh of fish prepares the purposes of active peptides compound |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103805665A (en) * | 2014-02-14 | 2014-05-21 | 青岛华科生物技术有限公司 | Preparation method of deep-sea fishskin collagen polypeptide |
| CN106319014A (en) * | 2016-10-14 | 2017-01-11 | 浙江双屿生物股份有限公司 | Extracting method for micro-molecule deep-sea fish polypeptides |
-
2017
- 2017-07-31 CN CN201710640809.4A patent/CN107397954A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103805665A (en) * | 2014-02-14 | 2014-05-21 | 青岛华科生物技术有限公司 | Preparation method of deep-sea fishskin collagen polypeptide |
| CN106319014A (en) * | 2016-10-14 | 2017-01-11 | 浙江双屿生物股份有限公司 | Extracting method for micro-molecule deep-sea fish polypeptides |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108077939A (en) * | 2017-11-29 | 2018-05-29 | 兰溪市沉默生物科技有限公司 | The deep-sea fish flesh of fish prepares the purposes of active peptides compound |
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Application publication date: 20171128 |