CN111995671A - Collagen peptide and application thereof - Google Patents
Collagen peptide and application thereof Download PDFInfo
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- CN111995671A CN111995671A CN202010927077.9A CN202010927077A CN111995671A CN 111995671 A CN111995671 A CN 111995671A CN 202010927077 A CN202010927077 A CN 202010927077A CN 111995671 A CN111995671 A CN 111995671A
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- collagen peptide
- melanoma cells
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- 102000008186 Collagen Human genes 0.000 title claims abstract description 48
- 108010035532 Collagen Proteins 0.000 title claims abstract description 48
- 229920001436 collagen Polymers 0.000 title claims abstract description 48
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 47
- 230000000694 effects Effects 0.000 claims abstract description 28
- 230000008099 melanin synthesis Effects 0.000 claims abstract description 11
- 239000002537 cosmetic Substances 0.000 claims abstract description 10
- 230000002087 whitening effect Effects 0.000 claims abstract description 10
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 239000006071 cream Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 239000002023 wood Substances 0.000 claims description 3
- 239000000516 sunscreening agent Substances 0.000 claims description 2
- 230000006862 enzymatic digestion Effects 0.000 claims 1
- 210000002966 serum Anatomy 0.000 claims 1
- 230000000475 sunscreen effect Effects 0.000 claims 1
- 201000001441 melanoma Diseases 0.000 abstract description 20
- 102000003425 Tyrosinase Human genes 0.000 abstract description 11
- 108060008724 Tyrosinase Proteins 0.000 abstract description 11
- 239000000284 extract Substances 0.000 abstract description 5
- 231100000956 nontoxicity Toxicity 0.000 abstract description 3
- 239000004480 active ingredient Substances 0.000 abstract 1
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 10
- 239000007788 liquid Substances 0.000 description 9
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 8
- 229910002092 carbon dioxide Inorganic materials 0.000 description 8
- 238000012258 culturing Methods 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000012980 RPMI-1640 medium Substances 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 239000001569 carbon dioxide Substances 0.000 description 4
- 238000002791 soaking Methods 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 230000003698 anagen phase Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 2
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 2
- 102000029816 Collagenase Human genes 0.000 description 2
- 108060005980 Collagenase Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000001976 enzyme digestion Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000413 hydrolysate Substances 0.000 description 2
- BJRNKVDFDLYUGJ-RMPHRYRLSA-N hydroquinone O-beta-D-glucopyranoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-RMPHRYRLSA-N 0.000 description 2
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- 238000000926 separation method Methods 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
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- 238000000108 ultra-filtration Methods 0.000 description 2
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- 108091005658 Basic proteases Proteins 0.000 description 1
- 102000016387 Pancreatic elastase Human genes 0.000 description 1
- 108010067372 Pancreatic elastase Proteins 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 229960000271 arbutin Drugs 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 208000027697 autoimmune lymphoproliferative syndrome due to CTLA4 haploinsuffiency Diseases 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
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- 238000005238 degreasing Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000385 effect on melanoma Effects 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- BEJNERDRQOWKJM-UHFFFAOYSA-N kojic acid Chemical compound OCC1=CC(=O)C(O)=CO1 BEJNERDRQOWKJM-UHFFFAOYSA-N 0.000 description 1
- 229960004705 kojic acid Drugs 0.000 description 1
- WZNJWVWKTVETCG-UHFFFAOYSA-N kojic acid Natural products OC(=O)C(N)CN1C=CC(=O)C(O)=C1 WZNJWVWKTVETCG-UHFFFAOYSA-N 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- BJRNKVDFDLYUGJ-UHFFFAOYSA-N p-hydroxyphenyl beta-D-alloside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-UHFFFAOYSA-N 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
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- 230000001954 sterilising effect Effects 0.000 description 1
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- 230000009967 tasteless effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
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- 230000035899 viability Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- 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
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/10—Process efficiency
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
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- Wood Science & Technology (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Veterinary Medicine (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Gastroenterology & Hepatology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dermatology (AREA)
- Cosmetics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a collagen peptide and application thereof. The collagen peptide is a Qinghai yak skin extract, has a molecular weight of less than 1000Da, and has the effect of inhibiting melanin synthesis. The natural collagen peptide has no toxicity, can inhibit the synthesis of melanin by reducing the activity of tyrosinase in melanoma cells, and can be used as an active ingredient for preparing whitening cosmetics.
Description
Technical Field
The invention belongs to the field of cosmetics, and particularly relates to a collagen peptide and application thereof.
Background
The cloud of the ancient times, namely the ugly with one white covering and three clown, shows that the pursuit of white skin of people from the ancient times to the present time is unchanged. Therefore, cosmetics with whitening efficacy are one of the mainstream of skin care products, and research and development of various large manufacturers are hot.
The principle of whitening is to achieve the skin whitening effect by inhibiting the activity of tyrosinase in the body and thus reducing the generation of melanin. At present, the whitening cosmetics added in the market comprise one or more of arbutin, kojic acid, ascorbic acid, cell growth factors, plant natural extracts and other raw materials, and although the effects of inhibiting tyrosinase activity can be achieved, certain side effects exist, certain damage is caused to skin, products are not stable and easy to inactivate, and the currently popular plant natural extracts also have the problems of low efficiency, high cost and the like. Therefore, the invention discloses a natural whitening product which has strong tyrosinase inhibitory activity, can reduce the generation of melanin, is easy to be absorbed by human bodies and has no toxic or side effect.
Disclosure of Invention
The invention aims to provide a collagen peptide and application thereof, wherein the collagen peptide has the effect of inhibiting melanin synthesis, has no toxicity, is beneficial to human body absorption, and has a good application prospect in the field of whitening cosmetics.
The purpose of the invention is realized by the following technical scheme.
A collagen peptide is characterized in that the collagen peptide is extracted from the skin of Qinghai Chaaida wood basin yak, and the molecular weight is less than 1000 Da; the collagen peptide has the effect of inhibiting melanin synthesis.
As a preferred technical scheme, the collagen peptide is extracted by a denaturation combined compound directional enzyme digestion technology.
As a preferred technical scheme, the yak is in a pure natural ecological environment, is cultivated at low temperature, low oxygen and strong light, and extracts high-quality collagen peptide from unique raw materials.
An application of the collagen peptide in the field of whitening cosmetics.
As a preferable technical scheme, the beauty cosmetics comprise essence, eye cream, sun cream and facial masks.
The invention has the beneficial effects that: the collagen peptide is naturally extracted from yaks, has high homology with collagen in human bodies, is beneficial to absorption of the human bodies and has no toxicity. Particularly, at the level of melanoma cells, the composition can obviously reduce the activity of tyrosinase, inhibit the generation of melanin, and has the effectiveness and safety of collagen peptide, so that the composition can be completely used for preparing whitening cosmetics.
Drawings
FIG. 1 is a graph showing the effect of the collagen peptides of the present invention on the viability of melanoma cells.
FIG. 2 is a graph showing the effect of the collagen peptide of the present invention on the synthesis of melanin by melanoma cells.
FIG. 3 is a graph showing the effect of the collagen peptide of the present invention on tyrosinase activity in melanoma cells.
Detailed Description
The present invention will be further described with reference to the following detailed description, which should be construed as illustrative only, and not limiting the scope of the invention, which is to be given the full breadth of the appended claims, and all changes that can be made by those skilled in the art and which are, therefore, intended to be embraced therein.
Example 1
The collagen peptide is extracted from the skin of Qinghai chai-da wood basin yak which is bred in a pure natural ecological environment with low temperature, low oxygen and strong light as a raw material by a composite directional enzyme digestion technology, has the effect of inhibiting melanin synthesis, and is prepared by the following specific steps:
(1) pretreatment: weighing fresh yak skin, unhairing, cutting into blocks of 1 × 1cm, adding soda with mass concentration of 5% into the blocks according to the mass ratio of 1:5kg/kg of liquid at room temperature, soaking for 8 hours, washing the yak skin with clear water to be neutral after soaking, then adding the yak skin into salt with mass concentration of 5% according to the mass ratio of 1:5kg/kg of liquid, soaking for 8 hours again, and washing the yak skin with clear water to be neutral after soaking to achieve the purposes of degreasing and impurity removal;
(2) homogenizing: fully homogenizing the pretreated yak skin by using a colloid mill to prepare yak skin homogenate;
(3) denaturation: adding 1:5kg/kg of pure water in parts by mass into the yak skin slurry obtained in the step (2), and carrying out heat preservation treatment at the temperature of 60-80 ℃ for 4-6 hours to obtain a yak skin denatured liquid;
(4) ultra-fine treatment: sieving the denatured liquid with a 200-mesh sieve, collecting the undersize liquid, and performing ultramicro refinement treatment by using a nano collider to ensure that the average particle size of the treated undersize liquid is less than 500nm to obtain a nano enzymatic hydrolysate;
(5) enzymolysis: adjusting the pH value of the nano enzymatic hydrolysate to 9 by adopting NaOH, adding 0.3g of alkaline protease with 20 ten thousand of enzyme activity and 0.03g of elastase with 10 ten thousand of enzyme activity, and carrying out enzymolysis at 50 ℃ for 4-6 h;
(6) centrifugal separation: and (3) boiling the enzymolysis liquid obtained in the step (5) at high temperature for 30min for enzyme deactivation and sterilization, centrifuging at 10000rpm to remove residues, and performing microfiltration to obtain the collagen peptidase enzymolysis liquid.
(7) Purifying with macroporous adsorption resin: passing the collagen peptidase hydrolyzed solution obtained after centrifugal separation in the step (6) through a pre-filled macroporous adsorption resin chromatographic column at a proper flow rate, eluting 1-4 column volumes by using clear water to remove impurities after sample loading is finished, eluting by using 55-80% ethanol water solution, and collecting the eluent to obtain a collagen peptide purified solution;
(8) and (3) concentrating under reduced pressure: concentrating the purified yak bone collagen peptide solution under reduced pressure until the solution has no alcohol taste, and recovering ethanol to obtain concentrated yak bone collagen peptide solution;
(9) and (3) ultrafiltration: performing 1KDa ultrafiltration membrane treatment on the concentrated solution in the step (8);
(10) and (3) drying: and (4) carrying out spray drying on the enzymolysis liquid treated in the last step to prepare the colorless and tasteless collagen peptide.
Example 2
The CCK8 method detects the effect of the collagen peptide on the activity of melanoma cells.
Melanoma cells B16-F10 in RPMI 1640 medium (containing 10% fetal bovine serum) at 5% CO2And culturing in a carbon dioxide incubator at the constant temperature of 37 ℃. Taking melanoma cells in exponential phase growth phase after passage, and adjusting cell density to be 2.5 multiplied by 104one/mL, inoculated in a 96-well plate at 100. mu.l/well at 37 ℃ in 5% CO2After culturing for 24 hours, the collagen peptide sample of the present invention was added (dissolved in RPMI 1640 culture solution and filtered through a 0.22 μm filter) to a final concentration of 0.2, 1.56, 3.12, 6.25, 12.5 mg/mL. After 48 days of culture, 100. mu.l of the culture medium was aspirated10% CCK8, culturing in a carbon dioxide incubator for 2h, detecting absorbance (OD value) at 450nm by using a multifunctional microplate reader, and finally calculating the cell survival rate of each group. Each experimental group was set to 5 replicates and the control group was added to the same volume of RPMI 1640 medium. The cell survival rate (%) was (OD experimental group to OD blank)/(OD control group to OD blank) x 100%, and the results are shown in fig. 1.
As can be seen from FIG. 1, the low concentration of the collagen peptide of the present invention had no significant effect on melanoma cells. Compared with a control group, the collagen peptide has no obvious influence on the activity of melanoma cells at the concentration of less than 6.25mg/mL, and the fact that the collagen peptide has no cytotoxicity is proved.
Example 3
The collagen peptide of the invention has the effect on the synthesis of melanin by melanoma cells.
Melanoma cells B16-F10 in RPMI 1640 medium (containing 10% fetal bovine serum) at 5% CO2And culturing in a carbon dioxide incubator at the constant temperature of 37 ℃. Taking melanoma cells in exponential phase growth phase after passage, and adjusting cell density to be 1 × 106one/mL, 100. mu.l/well in six well plates, 5% CO at 37 ℃2After culturing for 24 hours, the sample (the collagen peptide sample of the present invention was dissolved in DMEM and filtered through a 0.22 μm filter) was added to a final concentration of 0.2mg/mL or 2 mg/mL. After 48h of culture, the digested cells were resuspended in 1mL of medium and the cells counted. After counting, centrifuging, removing the culture medium, adding 1mL of 1mol/L NaOH solution containing 10% DMSO to lyse the cells, ultrasonically crushing for 1min, and carrying out water bath at 90 ℃ for 2 h. And (5) measuring the A value at 450nm of the microplate reader, and calculating the relative content of the melanin in the cells. In this experiment, a medium containing no collagen peptide of the present invention was set as a control, and 3 replicates were set for each group. The relative melanin synthesis content is calculated as follows:
the relative melanin synthesis content/%, which is [ (treated group a value/treated group cell density)/(control group a value/control group cell density) ] × 100%, is shown in fig. 2.
As shown in FIG. 2, the collagen peptide of the present invention significantly inhibited melanin synthesis in melanoma cells. Compared with a control group, the collagen peptide of the invention with the concentration of 0.2mg/mL and 2mg/mL respectively reduces the relative content of melanin in melanoma cells to 60.12 percent and 63.06 percent.
Example 4
Effect of the collagen peptides of the invention on tyrosinase activity in melanoma cells.
The invention relates to a tyrosinase activity detection kit (Beijing Solebao scientific and technological Co., Ltd.) for determining the tyrosinase activity in melanoma cells by using collagen peptide.
Melanoma cells B16-F10 in RPMI 1640 medium (containing 10% fetal bovine serum) at 5% CO2And culturing in a carbon dioxide incubator at the constant temperature of 37 ℃. Taking melanoma cells in exponential phase growth phase after passage, and adjusting cell density to be 1 × 106one/mL, 100. mu.l/well in six well plates, 5% CO at 37 ℃2After culturing for 24 hours, the samples (the collagen peptide samples of the present invention were dissolved in DMEM medium and filtered through a 0.22 μm filter) were added to give final concentrations of 0.05mg/mL, 0.2mg/mL, and 2 mg/mL. After 48h of culture, the digested cells were resuspended in 1mL of medium and the cells counted. And after counting, removing the culture medium by centrifugation, adding 1mL of kit extract, carrying out ultrasonic crushing, 12000g, centrifuging for 20 minutes at 4 ℃, taking the supernatant, and placing on ice for testing. In this experiment, a medium containing no collagen peptide of the present invention was set as a control, and 3 replicates were set for each group. The procedure was carried out as described and the results are shown in FIG. 3.
As can be seen from FIG. 3, the collagen peptide of the present invention significantly reduced tyrosinase activity in melanoma cells. Compared with a control group, the collagen peptide of the invention of 0.05mg/mL, 0.2mg/mL and 2mg/mL respectively reduces the relative activity of tyrosinase in melanoma cells to 62.16%, 61.70% and 76.29% under different concentrations.
Claims (5)
1. A collagen peptide is characterized in that the collagen peptide is extracted from the skin of Qinghai Chaaida wood basin yak, and the molecular weight is less than 1000 Da; the collagen peptide has the effect of inhibiting melanin synthesis.
2. The collagen peptide according to claim 1, wherein said collagen peptide is extracted by denaturing combined with complex directional enzymatic digestion.
3. The collagen peptide according to claim 1, wherein said Qinghai-Tibet plateau yak is a pure natural ecological environment, and is cultivated in low temperature, low oxygen and strong light.
4. Use of the collagen peptide according to claim 1 in the field of whitening cosmetics.
5. The use according to claim 4, wherein the cosmetic composition comprises a serum, an eye cream, a sunscreen cream, a mask.
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| CN202010927077.9A CN111995671B (en) | 2020-09-07 | 2020-09-07 | Collagen peptide and application thereof |
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| CN202010927077.9A CN111995671B (en) | 2020-09-07 | 2020-09-07 | Collagen peptide and application thereof |
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| CN111995671A true CN111995671A (en) | 2020-11-27 |
| CN111995671B CN111995671B (en) | 2023-06-20 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112574294A (en) * | 2020-12-30 | 2021-03-30 | 青海瑞肽生物科技有限公司 | Collagen peptide and preparation method and application thereof |
| CN113527428A (en) * | 2021-09-17 | 2021-10-22 | 渤海水产育苗(山东)有限公司 | Sea cucumber peptide and application and product thereof |
| CN114129712A (en) * | 2021-12-16 | 2022-03-04 | 天津知了大健康产业发展有限公司 | Whitening collagen peptide freeze-dried powder and preparation method thereof |
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| CN101353380A (en) * | 2008-05-20 | 2009-01-28 | 张丽丽 | Jellyfish origin immune activity enhanced collagen peptide, preparation and use thereof |
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| CN114129712A (en) * | 2021-12-16 | 2022-03-04 | 天津知了大健康产业发展有限公司 | Whitening collagen peptide freeze-dried powder and preparation method thereof |
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