CN108929381A - Skin emergency repair collagen tripeptide and preparation method thereof - Google Patents
Skin emergency repair collagen tripeptide and preparation method thereof Download PDFInfo
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- CN108929381A CN108929381A CN201810869989.8A CN201810869989A CN108929381A CN 108929381 A CN108929381 A CN 108929381A CN 201810869989 A CN201810869989 A CN 201810869989A CN 108929381 A CN108929381 A CN 108929381A
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- 102000008186 Collagen Human genes 0.000 title claims abstract description 67
- 108010035532 Collagen Proteins 0.000 title claims abstract description 67
- 229920001436 collagen Polymers 0.000 title claims abstract description 67
- 230000008439 repair process Effects 0.000 title claims abstract description 46
- 238000002360 preparation method Methods 0.000 title claims abstract description 41
- 108010010803 Gelatin Proteins 0.000 claims abstract description 60
- 239000008273 gelatin Substances 0.000 claims abstract description 60
- 229920000159 gelatin Polymers 0.000 claims abstract description 60
- 235000019322 gelatine Nutrition 0.000 claims abstract description 60
- 235000011852 gelatine desserts Nutrition 0.000 claims abstract description 60
- 241000237525 Mytilidae Species 0.000 claims abstract description 32
- 239000004365 Protease Substances 0.000 claims abstract description 29
- 238000000034 method Methods 0.000 claims abstract description 18
- 108091005804 Peptidases Proteins 0.000 claims abstract description 13
- 235000019419 proteases Nutrition 0.000 claims abstract description 12
- 108090000526 Papain Proteins 0.000 claims abstract description 11
- 235000006886 Zingiber officinale Nutrition 0.000 claims abstract description 11
- 235000008397 ginger Nutrition 0.000 claims abstract description 11
- 235000019834 papain Nutrition 0.000 claims abstract description 11
- 229940055729 papain Drugs 0.000 claims abstract description 11
- 241000196324 Embryophyta Species 0.000 claims abstract description 7
- 108010058314 rennet Proteins 0.000 claims abstract description 7
- 229940108461 rennet Drugs 0.000 claims abstract description 7
- 108010004032 Bromelains Proteins 0.000 claims abstract description 3
- 108090000270 Ficain Proteins 0.000 claims abstract description 3
- 235000019835 bromelain Nutrition 0.000 claims abstract description 3
- 235000019836 ficin Nutrition 0.000 claims abstract description 3
- POTUGHMKJGOKRI-UHFFFAOYSA-N ficin Chemical compound FI=CI=N POTUGHMKJGOKRI-UHFFFAOYSA-N 0.000 claims abstract description 3
- 244000273928 Zingiber officinale Species 0.000 claims abstract 2
- 239000000243 solution Substances 0.000 claims description 57
- 240000002853 Nelumbo nucifera Species 0.000 claims description 53
- 235000006508 Nelumbo nucifera Nutrition 0.000 claims description 53
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 46
- 235000013372 meat Nutrition 0.000 claims description 42
- 239000007788 liquid Substances 0.000 claims description 41
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 34
- 229920002472 Starch Polymers 0.000 claims description 31
- 235000019698 starch Nutrition 0.000 claims description 31
- 239000008107 starch Substances 0.000 claims description 31
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 29
- 239000007864 aqueous solution Substances 0.000 claims description 27
- 239000003431 cross linking reagent Substances 0.000 claims description 27
- 238000005406 washing Methods 0.000 claims description 23
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 21
- SYUXAJSOZXEFPP-UHFFFAOYSA-N glutin Natural products COc1c(O)cc2OC(=CC(=O)c2c1O)c3ccccc3OC4OC(CO)C(O)C(O)C4O SYUXAJSOZXEFPP-UHFFFAOYSA-N 0.000 claims description 21
- 235000011121 sodium hydroxide Nutrition 0.000 claims description 21
- 108090000712 Cathepsin B Proteins 0.000 claims description 20
- 102000004225 Cathepsin B Human genes 0.000 claims description 20
- 101710180012 Protease 7 Proteins 0.000 claims description 20
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 19
- 238000010792 warming Methods 0.000 claims description 18
- 108010089934 carbohydrase Proteins 0.000 claims description 16
- 239000012530 fluid Substances 0.000 claims description 16
- 239000000413 hydrolysate Substances 0.000 claims description 15
- RDOXTESZEPMUJZ-UHFFFAOYSA-N anisole Chemical group COC1=CC=CC=C1 RDOXTESZEPMUJZ-UHFFFAOYSA-N 0.000 claims description 13
- 102000035195 Peptidases Human genes 0.000 claims description 12
- 239000002253 acid Substances 0.000 claims description 12
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 11
- 229960001484 edetic acid Drugs 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 10
- 230000008569 process Effects 0.000 claims description 9
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 claims description 8
- 238000004140 cleaning Methods 0.000 claims description 8
- 239000000499 gel Substances 0.000 claims description 8
- UIHCLUNTQKBZGK-UHFFFAOYSA-N Methyl isobutyl ketone Natural products CCC(C)C(C)=O UIHCLUNTQKBZGK-UHFFFAOYSA-N 0.000 claims description 7
- 108010009736 Protein Hydrolysates Proteins 0.000 claims description 7
- 239000000654 additive Substances 0.000 claims description 7
- 230000000996 additive effect Effects 0.000 claims description 7
- 229940043265 methyl isobutyl ketone Drugs 0.000 claims description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 229910052723 transition metal Inorganic materials 0.000 claims description 5
- -1 transition metal salt Chemical class 0.000 claims description 5
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 claims description 4
- 239000002738 chelating agent Substances 0.000 claims description 4
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 4
- UZKWTJUDCOPSNM-UHFFFAOYSA-N methoxybenzene Substances CCCCOC=C UZKWTJUDCOPSNM-UHFFFAOYSA-N 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 3
- 239000003960 organic solvent Substances 0.000 claims description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 2
- 239000005711 Benzoic acid Substances 0.000 claims description 2
- 235000010233 benzoic acid Nutrition 0.000 claims description 2
- 235000019253 formic acid Nutrition 0.000 claims description 2
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 2
- 150000005217 methyl ethers Chemical class 0.000 claims description 2
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 claims 1
- XBBGWXVKPRUITN-UHFFFAOYSA-N acetic acid;cyclohexane-1,1-diamine;sodium Chemical compound [Na].CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O.NC1(N)CCCCC1 XBBGWXVKPRUITN-UHFFFAOYSA-N 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 239000000843 powder Substances 0.000 claims 1
- 238000010521 absorption reaction Methods 0.000 abstract description 15
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 12
- 229940088598 enzyme Drugs 0.000 abstract description 10
- 102000004190 Enzymes Human genes 0.000 abstract description 8
- 108090000790 Enzymes Proteins 0.000 abstract description 8
- 239000002994 raw material Substances 0.000 abstract description 8
- 230000007760 free radical scavenging Effects 0.000 abstract description 6
- 230000007062 hydrolysis Effects 0.000 abstract description 6
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 6
- 238000009472 formulation Methods 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract description 2
- 230000000474 nursing effect Effects 0.000 abstract description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract 1
- 210000003491 skin Anatomy 0.000 description 37
- 241000234314 Zingiber Species 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 210000001519 tissue Anatomy 0.000 description 9
- 230000000694 effects Effects 0.000 description 7
- 238000007605 air drying Methods 0.000 description 6
- 235000013305 food Nutrition 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 229910021586 Nickel(II) chloride Inorganic materials 0.000 description 4
- 239000003513 alkali Substances 0.000 description 4
- QMMRZOWCJAIUJA-UHFFFAOYSA-L nickel dichloride Chemical compound Cl[Ni]Cl QMMRZOWCJAIUJA-UHFFFAOYSA-L 0.000 description 4
- 238000005507 spraying Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Natural products CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 2
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 2
- 241000269978 Pleuronectiformes Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000013475 authorization Methods 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 235000015170 shellfish Nutrition 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- RNMCCPMYXUKHAZ-UHFFFAOYSA-N 2-[3,3-diamino-1,2,2-tris(carboxymethyl)cyclohexyl]acetic acid Chemical compound NC1(N)CCCC(CC(O)=O)(CC(O)=O)C1(CC(O)=O)CC(O)=O RNMCCPMYXUKHAZ-UHFFFAOYSA-N 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 241000276435 Gadus Species 0.000 description 1
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 208000001126 Keratosis Diseases 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001072261 Musculista senhousia Species 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 241000242583 Scyphozoa Species 0.000 description 1
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 208000030961 allergic reaction Diseases 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000002144 chemical decomposition reaction Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 125000005909 ethyl alcohol group Chemical group 0.000 description 1
- 125000003916 ethylene diamine group Chemical group 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 108010007119 flavourzyme Proteins 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001473 noxious effect Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
Classifications
-
- 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]
-
- 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
-
- 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
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Veterinary Medicine (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- Biochemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Genetics & Genomics (AREA)
- Dermatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Toxicology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Cosmetics (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention provides a kind of skin emergency repair collagen tripeptides and preparation method thereof, and the invention belongs to nursing skin formulation arts.The present invention selects Mytilidae animal gelatin as raw material, prepares collagen tripeptide by dual-enzyme hydrolysis method, wherein double enzymes are plant rennet, is selected from bromelain, papain, ficin and Ginger Protease.The present invention is also made that improvement to the preparation method of the preparation method of raw material and collagen peptide.Collagen tripeptide prepared by the present invention, molecule is small, absorbs fastly, has good percutaneous absorbability, 24 hours transdermal absorption factors are up to 3780 μ g/cm2, 30 minutes free radical scavenging activities reach as high as 86%, can be used for the emergency repair of skin.
Description
Technical field
The present invention relates to nursing skin formulation art, especially a kind of skin emergency repair collagen tripeptide and its preparation side
Method.
Background technique
Main component of the collagen as extracellular matrix is the typical fibers albumen in multicellular animals, accounts for body
The 25-30% of total protein content is the most abundant protein of content in animal, has special mechanical performance and biological nature.
In recent years, as that studies collagen deepens continuously, the application in medical domain, beauty culture and food industry is also got over
Come more extensive, collagen has been considered as one kind and has excellent performance, is widely used and the very high natural biological of Development volue at present
Substance.
Collagen polypeptide can be dissolved completely in water, and aqueous solution low-viscosity also has mobility, acid resistance under 60% high concentration
Good, without precipitating in the case where acid, alkali, high temperature resistance is good, and 200 DEG C of heating are also without precipitating.Since its molecular mass is smaller,
So being easier to be absorbed by the body.For collagen polypeptide compared with vegetable protein, absorption rate is higher, and degree of absorption can reach, and not have
There is allergic reaction, and the absorption of other protein in food can be promoted.
The production method of protein hydrolysate is divided into chemical degradation method and enzyme edman degradation Edman.Chemical method is with chemistry such as acid, alkali
Reagent promotes the skin chain of protein molecule to break to form small-molecule substance at a certain temperature.Acid system mostly uses hydrochloric acid, sulfuric acid etc.
Strong acid reacts at high temperature, strong reaction, equipment seriously corroded, and hydrolysis thoroughly, generates ispol, while in high temperature more
Lower tryptophan is destroyed completely.Alkali process hydrolysis can also make natural l-amino acid racemization, moreover it is possible to form noxious material, inanimate object benefit
With value, therefore should not use.
Arrived in the 1980s, with Enzymes Industry and food industry fast development, it has been found that enzymatic hydrolysis
Reaction temperature is low, and the reaction time is short, non-environmental-pollution.Product nutritive value is high, instant based on polypeptide and type free amino acid
Property it is good, be easy to human consumption's absorption, thus attention has focused on using on enzymic hydrolysates.
There are alkali protease, neutral proteinase, papain, trypsase, stomach egg using wider protease at present
White enzyme, flavor protease Flavourzyme, compound protease Protamex etc..
In the prior art, " Enzymatic Production of Flatfish Skin Collagen Oligopeptide " (food research and development, 2006,27 (8):
104-107.) selected works hydrolysis by novo flatfish skin, has obtained collagen peptide, and maximum degree of hydrolysis is 22.36%.Authorization
Notification number provides a kind of enzyme with ginger rhizome for the Chinese invention patent of 104540960 B of CN and obtains glue to decompose collagen
Former peptide.For another example, Authorization Notice No. is the Chinese invention patent of 101870995 B of CN, uses three step enzymatic isolation methods by raw material of fish-skin
Prepare collagen tripeptide.
By above-mentioned document it is found that the raw material digested in the prior art selects skin or bone or the fish of mammal mostly
Skin, rare report for using shell-fish as raw material.Shell-fish has as a kind of very common, widely distributed animal
The huge business potential developed into collagen peptide raw material.
Summary of the invention
The present invention provides a kind of collagen tripeptides and preparation method thereof of skin emergency repair, utilize Mytilidae animal system
Standby gelatin prepares collagen tripeptide by dual-enzyme hydrolysis method, effectively overcomes that collagen peptide raw material sources in the prior art are narrow to ask
Topic.The collagen tripeptide of skin emergency repair provided by the invention, molecule is small, absorbs fastly, there is the emergency repair of excellent
Skin effect, Transdermal absorption effect and radicals scavenging effect.
The present invention is achieved through the following technical solutions:
A kind of skin emergency repair collagen tripeptide, is prepared by Mytilidae animal gelatin.
Preferably, the Mytilidae animal is Xun Shi flesh clam.
Preferably, which is characterized in that the Mytilidae animal gelatin preparation step is as follows:
(1) it cleans: Mytilidae animal flesh is separated with shell, with 4-5 times of meat quality of water washing of Mytilidae animal, washing
It is taken out after being impregnated 1-2 hours with 4-5 times of meat quality of Mytilidae animal of water again after the completion spare；
(2) it pre-processes: the Mytilidae animal flesh after cleaning being homogenized with tissue refiner, Mytilidae animal meat quality is added
The water of 2-3 times of quality is heated to 80-100 DEG C, is kept for 10-30 minutes, obtains pretreatment fluid；
(3) carbohydrase is degraded: by pretreatment fluid acid solution adjusting pH value to 4.0-4.5, while adjusting the temperature to 60-
65 DEG C, carbohydrase is added and digests 2-3 hours, obtains enzymolysis liquid；
(4) enzyme-deactivating: being warming up to 95-100 DEG C for enzymolysis liquid, kept for 5-15 minutes, is cooled to 30-40 DEG C, centrifugal filtration,
Filter residue is removed, enzyme-deactivating liquid is obtained；
(5) gel forming: cross-linking agent solution is mixed with enzyme-deactivating liquid, is stood 10-12 hours at room temperature, is obtained Mytilidae
Animal gelatin；
The volume ratio of the cross-linking agent solution and enzyme-deactivating solution is 1:(4-5).
Preferably, the acid solution be 0.01-0.15mol/L hydrochloric acid solution, 0.01-0.5mol/L acetic acid aqueous solution,
0.01-0.5mol/L aqueous formic acid, 0.01-1.0mol/L benzoic acid aqueous solution, in 0.01-0.1mol/L aqueous citric acid solution
Any one.
Preferably, the cross-linking agent solution is the transition metal salt of (2-3): 1:(30-40): chelating agent: water by molar ratio
It is mixed to get；
The chelating agent is ethylenediamine, ethylenediamine tetra-acetic acid, triethylenetetraaminehexaacetic acid and cyclohexanediamine tetraacetic acid
Any one in sodium.
Preferably, the transition metal salt is Fe3+、Co2+And Ni2+Water-soluble inorganic salt in any one or it is more
Kind.
It is highly preferred that the transition metal salt is FeCl3And NiCl2Molar ratio is the mixed metal salt of 1:1.
Skin emergency repair collagen tripeptide preparation method, steps are as follows:
(1) water-soluble with hydrochloric acid solution or sodium hydroxide after heating Mytilidae animal gelatin 5-10 minutes at 70-80 DEG C
Liquid adjusts pH value to the optimum pH value of protease A, while adjusting the temperature to the optimum temperature of protease A, and protease A is added
Enzymatic hydrolysis 2-3 hour, then with 10-20wt% hydrochloric acid solution or 20-40wt% sodium hydrate aqueous solution adjusting pH value to Cathepsin B most
Suitable pH value, while the optimum temperature of Cathepsin B is adjusted the temperature to, Cathepsin B is added and digests 2-3 hours, obtains gelatin
Enzymolysis liquid；
The protease A and Cathepsin B is plant rennet, and protease A and Cathepsin B be not identical；
(2) enzymolysis liquid is warming up to 95-100 DEG C, is kept for 5-15 minutes, is cooled to 30-40 DEG C, centrifugal filtration takes supernatant
Liquid is spray-dried to get skin emergency repair collagen tripeptide.
It is highly preferred that skin emergency repair collagen tripeptide preparation method, steps are as follows:
(1) water-soluble with hydrochloric acid solution or sodium hydroxide after heating Mytilidae animal gelatin 5-10 minutes at 70-80 DEG C
Liquid adjusts pH value to the optimum pH value of protease A, while adjusting the temperature to the optimum temperature of protease A, and protease A is added
Enzymatic hydrolysis 2-3 hours, then pH value is adjusted to the optimum pH value of Cathepsin B with hydrochloric acid solution or sodium hydrate aqueous solution, it adjusts simultaneously
Temperature is saved to the optimum temperature of Cathepsin B, Cathepsin B is added and digests 2-3 hours, obtains glutin hydrolysate；
The protease A and Cathepsin B is plant rennet, and protease A and Cathepsin B be not identical；
(2) lotus root starch is added into glutin hydrolysate, stands 1-2 hours, is warming up to 95-100 DEG C, is kept for 5-15 minutes, it is cold
But to 30-40 DEG C, centrifugation takes supernatant, is spray-dried to get skin emergency repair collagen tripeptide；
The additive amount of the lotus root starch is the 5-20% of Mytilidae animal gelatin quality.
Preferably, the plant rennet is bromelain, papain, ficin or Zingibain
Enzyme.
Preferably, the lotus root starch preparation method is after saving lotus root 0.5-3 hours at -20 DEG C, to be added in refiner, add
Enter the organic solvent of 3-5 times of quality of lotus root, is homogenized 2-3 minutes, obtains homogenate, the acetone washing with 5 times of lotus root starch quality is primary, mistake
Filter, natural air drying is at room temperature to get lotus root starch；
The organic solvent is methyl phenyl ethers anisole, three fourth Methyl ethers or methylisobutylketone.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, and following embodiment is merely illustrative this
Invention, is not intended to limit the present invention.
Specifically, the raw material or equipment source used in the following embodiments is as follows:
Xun Shi flesh clam, Classification system are Musculus senhousei, are purchased from Shantou Anping road fish market.
Lotus root, Classification system are Nelumbo nucifera Gaertn, are purchased from the market Shantou Xia Shan.
Tissue refiner is the DS-1 type tissue refiner of Hangzhou Xu Qing Science and Technology Ltd. production.
Hydrochloric acid, No. CAS: 7647-01-0.
Citric acid, No. CAS: 77-92-9.
Sodium hydroxide, No. CAS: 1310-73-2.
Carbohydrase, 80,000 U/g albumen of enzyme activity, the production of Shandong Kang Qin Biotechnology Co., Ltd.
Papain, 500,000 U/g albumen of enzyme activity, the production of Shanghai Li Zhu east wind Biotechnology Co., Ltd.
Ginger Protease presses " research of Ginger Protease enzyme activity determination method " (food science and technology, 2010 (1): 277-
281.) a text preparation, and measuring Zingibain activity obtained according to the method for this article is 140,000 U/g albumen.
NiCl2, No. CAS: 7718-54-9.
FeCl3, No. CAS: 7705-08-0.
Ethylenediamine tetra-acetic acid, No. CAS: 60-00-4.
Methyl phenyl ethers anisole, No. CAS: 100-66-3.
Methylisobutylketone, No. CAS: 108-10-1.
Embodiment 1
A kind of skin emergency repair collagen tripeptide is prepared by Xun Shi flesh clam gelatin.
Xun Shi flesh clam gelatin preparation step is as follows:
(1) it cleans: Xun Shi flesh clam meat is separated with shell, with 5 times of quality of water washing of Xun Shi flesh clam meat, after the completion of washing
It is taken out after being impregnated 2 hours with 5 times of quality of Xun Shi flesh clam meat of water again spare；
(2) it pre-processes: the Xun Shi flesh clam meat after cleaning being homogenized with tissue refiner, is homogenized 3 points with 10000rpm revolving speed
The water of 2 times of quality of Xun Shi flesh clam meat quality is added in clock, is heated to 80 DEG C, is kept for 15 minutes, obtains pretreatment fluid；
(3) carbohydrase is degraded: by pretreatment fluid 0.02mol/L hydrochloric acid solution adjusting pH value to 4.2, while adjusting temperature
To 65 DEG C, the glucoamylase enzyme that Xun Shi flesh clam meat quality 3% is added is digested 3 hours, obtains enzymolysis liquid；
(4) enzyme-deactivating: being warming up to 100 DEG C for enzymolysis liquid, keep 10min, is cooled to 40 DEG C, and 4000rpm is centrifuged 10 minutes,
It takes supernatant to cross 200 meshes, removes filter residue, obtain enzyme-deactivating liquid；
(5) gel forming: cross-linking agent solution is mixed with enzyme-deactivating liquid, stands 12 hours at room temperature, and it is dynamic to obtain Mytilidae
Object gelatin；
The volume ratio of the cross-linking agent solution and enzyme-deactivating solution is 1:5.
The NiCl that the cross-linking agent solution is 3:1:36 by molar ratio2: ethylenediamine tetra-acetic acid: water is mixed to get；
Skin emergency repair collagen tripeptide preparation method, steps are as follows:
(1) after Xun Shi flesh clam gelatin being heated 6 minutes at 70 DEG C, with 30wt% sodium hydrate aqueous solution adjust pH value to
6.5, while temperature 60 C is adjusted, papain enzymolysis 2 hours of Xun Shi flesh clam gelatin quality 3% are added, then use 30wt%
Sodium hydrate aqueous solution adjusts pH value to 7.0, while adjusting the temperature to 50 DEG C, and the ginger of Xun Shi flesh clam gelatin quality 3% is added
Protease hydrolyzed 2 hours, obtain glutin hydrolysate；
(2) glutin hydrolysate is warming up to 100 DEG C, keeps 10min, be cooled to 30 DEG C, 5000rpm is centrifuged 5 minutes, is taken
Clear liquid, with 150 DEG C of inlet air temperature, 70 DEG C of leaving air temp spray drying are to get skin emergency repair collagen tripeptide.
Embodiment 2
A kind of skin emergency repair collagen tripeptide is prepared by Xun Shi flesh clam gelatin.
Xun Shi flesh clam gelatin preparation step is as follows:
(1) it cleans: Xun Shi flesh clam meat is separated with shell, with 5 times of quality of water washing of Xun Shi flesh clam meat, after the completion of washing
It is taken out after being impregnated 2 hours with 5 times of quality of Xun Shi flesh clam meat of water again spare；
(2) it pre-processes: the Xun Shi flesh clam meat after cleaning being homogenized with tissue refiner, is homogenized 3 points with 10000rpm revolving speed
The water of 2 times of quality of Xun Shi flesh clam meat quality is added in clock, is heated to 80 DEG C, is kept for 15 minutes, obtains pretreatment fluid；
(3) carbohydrase is degraded: by pretreatment fluid 0.02mol/L hydrochloric acid solution adjusting pH value to 4.2, while adjusting temperature
To 65 DEG C, the carbohydrase that Xun Shi flesh clam meat quality 3% is added is digested 3 hours, obtains enzymolysis liquid；
(4) enzyme-deactivating: being warming up to 100 DEG C for enzymolysis liquid, keep 10min, is cooled to 40 DEG C, and 4000rpm is centrifuged 10 minutes,
200 meshes are crossed, filter residue is removed, obtains enzyme-deactivating liquid；
(5) gel forming: cross-linking agent solution is mixed with enzyme-deactivating liquid, stands 12 hours at room temperature, and it is dynamic to obtain Mytilidae
Object gelatin；
The volume ratio of the cross-linking agent solution and enzyme-deactivating solution is 1:5.
The NiCl that the cross-linking agent solution is 3:1:36 by molar ratio2: ethylenediamine tetra-acetic acid: water is mixed to get；
Skin emergency repair collagen tripeptide preparation method, steps are as follows:
(1) after Xun Shi flesh clam gelatin being heated 6 minutes at 70 DEG C, with 30wt% sodium hydrate aqueous solution adjust pH value to
6.5, while temperature 60 C is adjusted, papain enzymolysis 2 hours of Xun Shi flesh clam gelatin quality 3% are added, then use 30wt%
Sodium hydrate aqueous solution adjusts pH value to 7.0, while adjusting the temperature to 50 DEG C, and the ginger of Xun Shi flesh clam gelatin quality 3% is added
Protease hydrolyzed 2 hours, obtain glutin hydrolysate；
(2) lotus root starch is added into glutin hydrolysate, stands 2 hours, glutin hydrolysate is warming up to 100 DEG C, is kept
10min is cooled to 30 DEG C, and 5000rpm is centrifuged 5 minutes, takes supernatant, and with 150 DEG C of inlet air temperature, 70 DEG C of leaving air temp spraying dry
It is dry to get skin emergency repair collagen tripeptide；
The additive amount of the lotus root starch is the 10% of Xun Shi flesh clam gelatin quality；
Lotus root starch preparation method is after saving lotus root 2 hours at -20 DEG C, and slice is added in refiner, and 3 times of quality of lotus root are added
Methyl phenyl ethers anisole, 10000rpm be homogenized 3 minutes, obtain homogenate, the acetone washing with 5 times of quality of lotus root is primary, cross 1000 meshes, institute
Solid at room temperature natural air drying to get lotus root starch.
Embodiment 3
A kind of skin emergency repair collagen tripeptide is prepared by Xun Shi flesh clam gelatin.
Xun Shi flesh clam gelatin preparation step is as follows:
(1) it cleans: Xun Shi flesh clam meat is separated with shell, with 5 times of quality of water washing of Xun Shi flesh clam meat, after the completion of washing
It is taken out after being impregnated 2 hours with 5 times of quality of Xun Shi flesh clam meat of water again spare；
(2) it pre-processes: the Xun Shi flesh clam meat after cleaning being homogenized with tissue refiner, is homogenized 3 points with 10000rpm revolving speed
The water of 2 times of quality of Xun Shi flesh clam meat quality is added in clock, is heated to 80 DEG C, is kept for 15 minutes, obtains pretreatment fluid；
(3) carbohydrase is degraded: by pretreatment fluid 0.02mol/L hydrochloric acid solution adjusting pH value to 4.2, while adjusting temperature
To 65 DEG C, the carbohydrase that Xun Shi flesh clam meat quality 3% is added is digested 3 hours, obtains enzymolysis liquid；
(4) enzyme-deactivating: being warming up to 100 DEG C for enzymolysis liquid, keep 10min, is cooled to 40 DEG C, and 4000rpm is centrifuged 10 minutes,
200 meshes are crossed, filter residue is removed, obtains enzyme-deactivating liquid；
(5) gel forming: cross-linking agent solution is mixed with enzyme-deactivating liquid, stands 12 hours at room temperature, and it is dynamic to obtain Mytilidae
Object gelatin；
The volume ratio of the cross-linking agent solution and enzyme-deactivating solution is 1:5.
The NiCl that the cross-linking agent solution is 3:1:36 by molar ratio2: ethylenediamine tetra-acetic acid: water is mixed to get；
Skin emergency repair collagen tripeptide preparation method, steps are as follows:
(1) after Xun Shi flesh clam gelatin being heated 6 minutes at 70 DEG C, with 30wt% sodium hydrate aqueous solution adjust pH value to
6.5, while temperature 60 C is adjusted, papain enzymolysis 2 hours of Xun Shi flesh clam gelatin quality 3% are added, then use 30wt%
Sodium hydrate aqueous solution adjusts pH value to 7.0, while adjusting the temperature to 50 DEG C, and the ginger of Xun Shi flesh clam gelatin quality 3% is added
Protease hydrolyzed 2 hours, obtain glutin hydrolysate；
(2) lotus root starch is added into glutin hydrolysate, stands 2 hours, glutin hydrolysate is warming up to 100 DEG C, is kept
10min is cooled to 30 DEG C, and 5000rpm is centrifuged 5 minutes, takes supernatant, and with 150 DEG C of inlet air temperature, 70 DEG C of leaving air temp spraying dry
It is dry to get skin emergency repair collagen tripeptide；
The additive amount of the lotus root starch is the 10% of Xun Shi flesh clam gelatin quality；
Lotus root starch preparation method is after saving lotus root 2 hours at -20 DEG C, and slice is added in refiner, and 3 times of quality of lotus root are added
Methylisobutylketone, 10000rpm be homogenized 3 minutes, obtain homogenate, the acetone washing with 5 times of quality of lotus root is primary, cross 1000 mesh
Sieve, obtained solid at room temperature natural air drying to get lotus root starch.
Embodiment 4
A kind of skin emergency repair collagen tripeptide is prepared by Xun Shi flesh clam gelatin.
Xun Shi flesh clam gelatin preparation step is as follows:
(1) it cleans: Xun Shi flesh clam meat is separated with shell, with 5 times of quality of water washing of Xun Shi flesh clam meat, after the completion of washing
It is taken out after being impregnated 2 hours with 5 times of quality of Xun Shi flesh clam meat of water again spare；
(2) it pre-processes: the Xun Shi flesh clam meat after cleaning being homogenized with tissue refiner, is homogenized 3 points with 10000rpm revolving speed
The water of 2 times of quality of Xun Shi flesh clam meat quality is added in clock, is heated to 80 DEG C, is kept for 15 minutes, obtains pretreatment fluid；
(3) carbohydrase is degraded: pretreatment fluid 0.05mol/L aqueous citric acid solution being adjusted pH value to 4.2, is adjusted simultaneously
To 65 DEG C, the carbohydrase that Xun Shi flesh clam meat quality 3% is added digests 3 hours temperature, obtains enzymolysis liquid；
(4) enzyme-deactivating: being warming up to 100 DEG C for enzymolysis liquid, keep 10min, is cooled to 40 DEG C, and 4000rpm is centrifuged 10 minutes,
200 meshes are crossed, filter residue is removed, obtains enzyme-deactivating liquid；
(5) gel forming: cross-linking agent solution is mixed with enzyme-deactivating liquid, stands 12 hours at room temperature, and it is dynamic to obtain Mytilidae
Object gelatin；
The volume ratio of the cross-linking agent solution and enzyme-deactivating solution is 1:5.
The NiCl that the cross-linking agent solution is 3:1:36 by molar ratio2: ethylenediamine tetra-acetic acid: water is mixed to get；
Skin emergency repair collagen tripeptide preparation method, steps are as follows:
(1) after Xun Shi flesh clam gelatin being heated 6 minutes at 70 DEG C, with 30wt% sodium hydrate aqueous solution adjust pH value to
6.5, while temperature 60 C is adjusted, papain enzymolysis 2 hours of Xun Shi flesh clam gelatin quality 3% are added, then use 30wt%
Sodium hydrate aqueous solution adjusts pH value to 7.0, while adjusting the temperature to 50 DEG C, and the ginger of Xun Shi flesh clam gelatin quality 3% is added
Protease hydrolyzed 2 hours, obtain glutin hydrolysate；
(2) lotus root starch is added into enzymolysis liquid, stands 2 hours, glutin hydrolysate is warming up to 100 DEG C, keeps 10min, it is cold
But to 30 DEG C, 5000rpm is centrifuged 5 minutes, takes supernatant, with 150 DEG C of inlet air temperature, 70 DEG C of leaving air temp spray drying to get
Skin emergency repair collagen tripeptide；
The additive amount of the lotus root starch is the 10% of Xun Shi flesh clam gelatin quality；Lotus root starch preparation method be by lotus root at -20 DEG C
After saving 2 hours, slice is added in refiner, and the methylisobutylketone of 3 times of quality of lotus root is added, and 10000rpm is homogenized 3 minutes, obtains
Homogenate, the acetone washing with 5 times of quality of lotus root is primary, cross 1000 meshes, obtained solid at room temperature natural air drying to get lotus root starch.
Embodiment 5
A kind of skin emergency repair collagen tripeptide is prepared by Xun Shi flesh clam gelatin.
Xun Shi flesh clam gelatin preparation step is as follows:
(1) it cleans: Xun Shi flesh clam meat is separated with shell, with 5 times of quality of water washing of Xun Shi flesh clam meat, after the completion of washing
It is taken out after being impregnated 2 hours with 5 times of quality of Xun Shi flesh clam meat of water again spare；
(2) it pre-processes: the Xun Shi flesh clam meat after cleaning being homogenized with tissue refiner, is homogenized 3 points with 10000rpm revolving speed
The water of 2 times of quality of Xun Shi flesh clam meat quality is added in clock, is heated to 80 DEG C, is kept for 15 minutes, obtains pretreatment fluid；
(3) carbohydrase is degraded: pretreatment fluid 0.05mol/L aqueous citric acid solution being adjusted pH value to 4.2, is adjusted simultaneously
To 65 DEG C, the carbohydrase that Xun Shi flesh clam meat quality 3% is added digests 3 hours temperature, obtains enzymolysis liquid；
(4) enzyme-deactivating: being warming up to 100 DEG C for enzymolysis liquid, keep 10min, is cooled to 40 DEG C, and 4000rpm is centrifuged 10 minutes,
200 meshes are crossed, filter residue is removed, obtains enzyme-deactivating liquid；
(5) gel forming: cross-linking agent solution is mixed with enzyme-deactivating liquid, stands 12 hours at room temperature, and it is dynamic to obtain Mytilidae
Object gelatin；
The volume ratio of the cross-linking agent solution and enzyme-deactivating solution is 1:5.
The FeCl that the cross-linking agent solution is 3:1:36 by molar ratio3: ethylenediamine tetra-acetic acid: water is mixed to get；
Skin emergency repair collagen tripeptide preparation method, steps are as follows:
(1) after Xun Shi flesh clam gelatin being heated 6 minutes at 70 DEG C, with 30wt% sodium hydrate aqueous solution adjust pH value to
6.5, while temperature 60 C is adjusted, papain enzymolysis 2 hours of Xun Shi flesh clam gelatin quality 3% are added, then use 30wt%
Sodium hydrate aqueous solution adjusts pH value to 7.0, while adjusting the temperature to 50 DEG C, and the ginger of Xun Shi flesh clam gelatin quality 3% is added
Protease hydrolyzed 2 hours, obtain glutin hydrolysate；
(2) lotus root starch is added into glutin hydrolysate, stands 2 hours, glutin hydrolysate is warming up to 100 DEG C, is kept
10min is cooled to 30 DEG C, and 5000rpm is centrifuged 5 minutes, takes supernatant, and with 150 DEG C of inlet air temperature, 70 DEG C of leaving air temp spraying dry
It is dry to get skin emergency repair collagen tripeptide；
The additive amount of the lotus root starch is the 10% of Xun Shi flesh clam gelatin quality；
Lotus root starch preparation method is after saving lotus root 2 hours at -20 DEG C, and slice is added in refiner, and 3 times of quality of lotus root are added
Methylisobutylketone, 10000rpm be homogenized 3 minutes, obtain homogenate, the acetone washing with 5 times of quality of lotus root is primary, cross 1000 mesh
Sieve, obtained solid at room temperature natural air drying to get lotus root starch.
Embodiment 6
A kind of skin emergency repair collagen tripeptide is prepared by Xun Shi flesh clam gelatin.
Xun Shi flesh clam gelatin preparation step is as follows:
(1) it cleans: Xun Shi flesh clam meat is separated with shell, with 5 times of quality of water washing of Xun Shi flesh clam meat, after the completion of washing
It is taken out after being impregnated 2 hours with 5 times of quality of Xun Shi flesh clam meat of water again spare；
(2) it pre-processes: the Xun Shi flesh clam meat after cleaning being homogenized with tissue refiner, is homogenized 3 points with 10000rpm revolving speed
The water of 2 times of quality of Xun Shi flesh clam meat quality is added in clock, is heated to 80 DEG C, is kept for 15 minutes, obtains pretreatment fluid；
(3) carbohydrase is degraded: pretreatment fluid 0.05mol/L aqueous citric acid solution being adjusted pH value to 4.2, is adjusted simultaneously
To 65 DEG C, the carbohydrase that Xun Shi flesh clam meat quality 3% is added digests 3 hours temperature, obtains enzymolysis liquid；
(4) enzyme-deactivating: being warming up to 100 DEG C for enzymolysis liquid, keep 10min, is cooled to 40 DEG C, and 4000rpm is centrifuged 10 minutes,
200 meshes are crossed, filter residue is removed, obtains enzyme-deactivating liquid；
(5) gel forming: cross-linking agent solution is mixed with enzyme-deactivating liquid, stands 12 hours at room temperature, and it is dynamic to obtain Mytilidae
Object gelatin；
The volume ratio of the cross-linking agent solution and enzyme-deactivating solution is 1:5.
The FeCl that the cross-linking agent solution is 1.5:1.5:1:36 by molar ratio3: NiCl2: ethylenediamine tetra-acetic acid: water mixing
It obtains；
Skin emergency repair collagen tripeptide preparation method, steps are as follows:
(1) after Xun Shi flesh clam gelatin being heated 6 minutes at 70 DEG C, with 30wt% sodium hydrate aqueous solution adjust pH value to
6.5, while temperature 60 C is adjusted, papain enzymolysis 2 hours of Xun Shi flesh clam gelatin quality 3% are added, then use 30wt%
Sodium hydrate aqueous solution adjusts pH value to 7.0, while adjusting the temperature to 50 DEG C, and the ginger of Xun Shi flesh clam gelatin quality 3% is added
Protease hydrolyzed 2 hours, obtain glutin hydrolysate；
(2) lotus root starch is added into glutin hydrolysate, stands 2 hours, glutin hydrolysate is warming up to 100 DEG C, is kept
10min is cooled to 30 DEG C, and 5000rpm is centrifuged 5 minutes, takes supernatant, and with 150 DEG C of inlet air temperature, 70 DEG C of leaving air temp spraying dry
It is dry to get skin emergency repair collagen tripeptide；
The additive amount of the lotus root starch is the 10% of Xun Shi flesh clam gelatin quality；
Lotus root starch preparation method is after saving lotus root 2 hours at -20 DEG C, and slice is added in refiner, and 3 times of quality of lotus root are added
Methylisobutylketone, 10000rpm be homogenized 3 minutes, obtain homogenate, the acetone washing with 5 times of quality of lotus root is primary, cross 1000 mesh
Sieve, obtained solid at room temperature natural air drying to get lotus root starch.
Test case 1
Transdermal absorption test
Due to only could really be absorbed by the body simultaneously after collagen peptide reaches skin corium through keratoderma, epidermis
It can reflect for repairing skin, therefore adding up transit dose by sample unit area in the Transdermal absorption measurements determination unit time
Its fixed skin emergency repair function.
Test method reference " the Transdermal absorption performance research of jellyfish, ox bone and gadus collagen peptide " (daily chemical industry,
2017 (3): 164-167.) collagen peptide absorption test in a text.With the embodiment 1-6 skin emergency repair being prepared
The aqueous solution that collagen tripeptide is configured to 10g/L respectively carries out Transdermal absorption test, and test result is as shown in table 1.
1 Transdermal absorption test result of table
As seen from the above table, emergency repair collagen tripeptide provided by the invention has preferable Transdermal absorption effect, when making
It is added to lotus root starch (embodiment 2) when during standby, unit area can be effectively promoted and add up transmitance, furthermore Xun Shi flesh clam gelatin
The crosslinking agent added in preparation process finally also will affect Transdermal absorption efficiency, the FeCl for being 1.5:1.5:1 by molar ratio3:
NiCl2: the gelatin preparation of cross-linking agent solution preparation is prepared in the water that 5 times of metal salt qualities are dissolved in after ethylenediamine tetra-acetic acid mixing
Made of collagen tripeptide (embodiment 6) have best Transdermal absorption effect.
Test case 2
Antioxidation in vitro test
Examination on experimental operation is as follows: preparing the DPPH titer of 250 μm of ol/L, is kept in dark place at 4 DEG C, matching while using is matched
Collagen tripeptide aqueous solution prepared by the embodiment 1-6 that concentration processed is 12mg/mL, takes 2mL to mix with isometric DPPH titer,
It is protected from light 30 minutes, absorbance A is measured at 515nm1, collagen tripeptide aqueous solution is replaced with ethyl alcohol, measures and inhales at 515nm
Luminosity A2, three peptide solutions (2mL collagen tripeptide aqueous solution+2mL ethyl alcohol) 515nm go out measure absorbance A3.Free radical scavenging ability
It is calculated as follows:
Test result is as shown in table 2.
2 DPPH free radical scavenging activity of table
As seen from the above table, collagen tripeptide provided by the invention has higher free radical scavenging activity, and addition lotus root starch changes
Acid solution and change crosslinking agent when prepared by gelatin can all influence free radical scavenging activity, wherein by the preparation of 6 method of embodiment
Collagen tripeptide has optimal 30 minutes free radical scavenging activities, up to 86%.
Claims (10)
1. a kind of skin emergency repair collagen tripeptide, which is characterized in that be prepared by Mytilidae animal gelatin.
2. skin emergency repair collagen tripeptide according to claim 1, which is characterized in that the Mytilidae animal is
Xun Shi flesh clam.
3. skin emergency repair collagen tripeptide according to claim 1, which is characterized in that the Mytilidae animal gelatin
Preparation step is as follows:
(1) it cleans: Mytilidae animal flesh is separated with shell, with 4-5 times of meat quality of water washing of Mytilidae animal, washing is completed
It is taken out after being impregnated 1-2 hours with 4-5 times of meat quality of Mytilidae animal of water again afterwards spare；
(2) it pre-processes: the Mytilidae animal flesh after cleaning being homogenized with tissue refiner, Mytilidae animal meat quality 2-3 is added
The water of times quality is heated to 80-100 DEG C, is kept for 10-30 minutes, obtains pretreatment fluid；
(3) carbohydrase is degraded: by pretreatment fluid acid solution adjusting pH value to 4.0-4.5, while adjusting the temperature to 60-65
DEG C, carbohydrase is added and digests 2-3 hours, obtains enzymolysis liquid；
(4) enzyme-deactivating: being warming up to 95-100 DEG C for enzymolysis liquid, kept for 5-15 minutes, is cooled to 30-40 DEG C, and centrifugation, filtering remove
Filter residue is removed, enzyme-deactivating liquid is obtained；
(5) gel forming: cross-linking agent solution is mixed with enzyme-deactivating liquid, is stood 10-12 hours at room temperature, is obtained Mytilidae animal
Gelatin；
The volume ratio of the cross-linking agent solution and enzyme-deactivating solution is 1:(4-5).
4. skin emergency repair collagen tripeptide according to claim 3, which is characterized in that the acid solution is
0.01-0.5mol/L acetic acid aqueous solution, 0.01-0.5mol/L aqueous formic acid, 0.01-1.0mol/L benzoic acid aqueous solution,
Any one in 0.01-0.1mol/L aqueous citric acid solution.
5. skin emergency repair collagen tripeptide according to claim 3, which is characterized in that the cross-linking agent solution by mole
Than the transition metal salt for (2-3): 1:(30-40): chelating agent: water is mixed to get；
The chelating agent is in ethylenediamine, ethylenediamine tetra-acetic acid, triethylenetetraaminehexaacetic acid and cyclohexanediamine tetraacetic acid sodium
Any one.
6. skin emergency repair collagen tripeptide according to claim 5, which is characterized in that the transition metal salt is Fe3 +、Co2+And Ni2+Water-soluble inorganic salt in any one or more.
7. skin emergency repair collagen tripeptide preparation method as claimed in any one of claims 1 to 6, which is characterized in that preparation
Steps are as follows:
(1) after heating Mytilidae animal gelatin 5-10 minutes at 70-80 DEG C, with hydrochloric acid solution or sodium hydrate aqueous solution tune
PH value is saved to the optimum pH value of protease A, while adjusting the temperature to the optimum temperature of protease A, protease A enzymatic hydrolysis is added
2-3 hours, then pH value is adjusted to Cathepsin B optimum with 10-20wt% hydrochloric acid solution or 20-40wt% sodium hydrate aqueous solution
PH value, while adjusting the temperature to the optimum temperature of Cathepsin B, Cathepsin B be added and digests 2-3 hour, obtain gelatin and digest
Liquid；
The protease A and Cathepsin B is plant rennet, and protease A and Cathepsin B be not identical；
(2) glutin hydrolysate is warming up to 95-100 DEG C, is kept for 5-15 minutes, is cooled to 30-40 DEG C, centrifugal filtration takes supernatant
Liquid is spray-dried to get skin emergency repair collagen tripeptide.
8. skin emergency repair collagen tripeptide preparation method as claimed in any one of claims 1 to 6, which is characterized in that preparation
Steps are as follows:
(1) after heating Mytilidae animal gelatin 5-10 minutes at 70-80 DEG C, with hydrochloric acid solution or sodium hydrate aqueous solution tune
PH value is saved to the optimum pH value of protease A, while adjusting the temperature to the optimum temperature of protease A, protease A enzymatic hydrolysis is added
2-3 hours, then pH value is adjusted to the optimum pH value of Cathepsin B with hydrochloric acid solution or sodium hydrate aqueous solution, while adjusting temperature
Degree is added Cathepsin B and digests 2-3 hours, obtain glutin hydrolysate to the optimum temperature of Cathepsin B；
The protease A and Cathepsin B is plant rennet, and protease A and Cathepsin B be not identical；
(2) lotus root starch is added into glutin hydrolysate, stands 1-2 hours, is warming up to 95-100 DEG C, is kept for 3-5 hours, is cooled to
30-40 DEG C, centrifugation takes supernatant, is spray-dried to get skin emergency repair collagen tripeptide；
The additive amount of the lotus root starch is the 5-20% of Mytilidae animal gelatin quality.
9. according to claim 7 or 8 state skin emergency repair collagen tripeptide preparation method, which is characterized in that described
Plant rennet be bromelain, papain, ficin or Ginger Protease.
10. according to claim 8 state skin emergency repair collagen tripeptide preparation method, which is characterized in that the lotus root
Powder, preparation method thereof is after saving lotus root 0.5-3 hours at -20 DEG C, to be added in refiner, and the organic molten of 3-5 times of quality of lotus root is added
Agent is homogenized 2-3 minutes, obtains homogenate, and the acetone washing with 5 times of lotus root starch quality is primary, filtering, and obtained solid is natural at room temperature
It air-dries to get lotus root starch；
The organic solvent is methyl phenyl ethers anisole, three fourth Methyl ethers or methylisobutylketone.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100004183A1 (en) * | 2005-10-13 | 2010-01-07 | Gu Jennifer L | Mineral collagen chelates and methods of making and using same |
| CN101870995A (en) * | 2009-04-21 | 2010-10-27 | 陈栋梁 | Process for preparing collagen tripeptide |
| CN103243144B (en) * | 2013-05-27 | 2014-05-28 | 天津益丽康生物科技有限公司 | Collagen powder rich in collagen tripeptide and preparation method thereof |
| CN104540960A (en) * | 2012-07-25 | 2015-04-22 | 株式会社日皮 | Collagen peptide composition production method, DPP-4 inhibitor, and antihyperglycemic agent |
| CN106752975A (en) * | 2016-12-06 | 2017-05-31 | 集美大学 | The preparation method of fishskin gelatin |
| CN107815483A (en) * | 2017-12-21 | 2018-03-20 | 大连深蓝肽科技研发有限公司 | The method that mussel oligopeptide is prepared using mussel fresh meat |
-
2018
- 2018-08-02 CN CN201810869989.8A patent/CN108929381B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100004183A1 (en) * | 2005-10-13 | 2010-01-07 | Gu Jennifer L | Mineral collagen chelates and methods of making and using same |
| CN101870995A (en) * | 2009-04-21 | 2010-10-27 | 陈栋梁 | Process for preparing collagen tripeptide |
| CN104540960A (en) * | 2012-07-25 | 2015-04-22 | 株式会社日皮 | Collagen peptide composition production method, DPP-4 inhibitor, and antihyperglycemic agent |
| CN103243144B (en) * | 2013-05-27 | 2014-05-28 | 天津益丽康生物科技有限公司 | Collagen powder rich in collagen tripeptide and preparation method thereof |
| CN106752975A (en) * | 2016-12-06 | 2017-05-31 | 集美大学 | The preparation method of fishskin gelatin |
| CN107815483A (en) * | 2017-12-21 | 2018-03-20 | 大连深蓝肽科技研发有限公司 | The method that mussel oligopeptide is prepared using mussel fresh meat |
Non-Patent Citations (11)
| Title |
|---|
| F. RODRÍGUEZ等: "Collagen extraction from mussel byssus: a new marine collagen source with physicochemical properties of industrial interest", 《JOURNAL OF FOOD SCIENCE AND TECHNOLOGY》 * |
| YOUNG CHAN CHOI等: "Human gelatin tissue-adhesive hydrogels prepared by enzyme-mediated biosynthesis of DOPA and Fe3+ ion", 《JOURNAL OF MATERIALS CHEMISTRY B》 * |
| 宋芹等: "一种胶原蛋白寡肽体外透皮吸收性能研究", 《成都大学学报》 * |
| 常映明等: "《抗衰老漫谈》", 30 November 2017, 中国医药科技出版社 * |
| 李亚楠等: "胶原三肽生物活性及其应用研究进展", 《食品工业科技》 * |
| 桑楚: "《优雅女人养成手册》", 30 April 2015, 中国华侨出版社 * |
| 罗光乾: "《驻颜革命》", 31 January 2009, 京华出版社 * |
| 胡建平: "《鱼胶原蛋白的开发与利用》", 31 March 2014, 四川大学出版社 * |
| 范继业等: "《生物化学》", 31 August 2017, 中国轻工业出版社 * |
| 陈健等: "《食品化学原理》", 28 February 2015, 华南理工大学出版社 * |
| 食物蛋白质控制酶解技术: "《食物蛋白质控制酶解技术》", 30 June 2018, 中国轻工业出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111494233A (en) * | 2020-06-02 | 2020-08-07 | 康珞生物科技（武汉）有限公司 | Essence with anti-aging effect |
| CN111494233B (en) * | 2020-06-02 | 2023-07-14 | 康珞生物科技（武汉）有限公司 | Preparation method and application of collagen short peptide liposome |
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