CN117357427A - Application of recombinant spider silk protein - Google Patents
Application of recombinant spider silk protein Download PDFInfo
- Publication number
- CN117357427A CN117357427A CN202311669459.6A CN202311669459A CN117357427A CN 117357427 A CN117357427 A CN 117357427A CN 202311669459 A CN202311669459 A CN 202311669459A CN 117357427 A CN117357427 A CN 117357427A
- Authority
- CN
- China
- Prior art keywords
- skin
- spider silk
- silk protein
- recombinant spider
- product
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 86
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 81
- 229920001872 Spider silk Polymers 0.000 title claims abstract description 77
- 210000004209 hair Anatomy 0.000 claims abstract description 54
- 201000004624 Dermatitis Diseases 0.000 claims abstract description 22
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 19
- 238000002360 preparation method Methods 0.000 claims abstract description 19
- 230000037303 wrinkles Effects 0.000 claims abstract description 15
- 239000002537 cosmetic Substances 0.000 claims abstract description 10
- 230000001153 anti-wrinkle effect Effects 0.000 claims abstract description 6
- 238000005406 washing Methods 0.000 claims abstract description 6
- 230000000694 effects Effects 0.000 claims description 42
- 108010003272 Hyaluronate lyase Proteins 0.000 claims description 16
- 102000001974 Hyaluronidases Human genes 0.000 claims description 16
- 229960002773 hyaluronidase Drugs 0.000 claims description 16
- 102000016387 Pancreatic elastase Human genes 0.000 claims description 11
- 108010067372 Pancreatic elastase Proteins 0.000 claims description 11
- 239000000463 material Substances 0.000 claims description 10
- 230000002401 inhibitory effect Effects 0.000 claims description 7
- 239000000654 additive Substances 0.000 claims description 6
- 230000000996 additive effect Effects 0.000 claims description 6
- 239000002453 shampoo Substances 0.000 claims description 4
- 239000000490 cosmetic additive Substances 0.000 claims description 2
- 230000002708 enhancing effect Effects 0.000 claims description 2
- 206010040954 Skin wrinkling Diseases 0.000 claims 3
- 238000011160 research Methods 0.000 abstract description 3
- 230000003752 improving hair Effects 0.000 abstract description 2
- 210000003491 skin Anatomy 0.000 description 49
- 239000000047 product Substances 0.000 description 38
- 239000000243 solution Substances 0.000 description 26
- 239000000523 sample Substances 0.000 description 25
- 108010022355 Fibroins Proteins 0.000 description 20
- 230000005764 inhibitory process Effects 0.000 description 13
- 239000007788 liquid Substances 0.000 description 13
- 238000012360 testing method Methods 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- 239000012460 protein solution Substances 0.000 description 10
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 206010020751 Hypersensitivity Diseases 0.000 description 7
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 7
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 239000013641 positive control Substances 0.000 description 7
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 6
- 239000012634 fragment Substances 0.000 description 6
- 241001052560 Thallis Species 0.000 description 5
- 230000007815 allergy Effects 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 238000001976 enzyme digestion Methods 0.000 description 5
- 239000013612 plasmid Substances 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000012795 verification Methods 0.000 description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 208000026935 allergic disease Diseases 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000000502 dialysis Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 229930027917 kanamycin Natural products 0.000 description 4
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 4
- 229960000318 kanamycin Drugs 0.000 description 4
- 229930182823 kanamycin A Natural products 0.000 description 4
- 230000009759 skin aging Effects 0.000 description 4
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 108010088751 Albumins Proteins 0.000 description 3
- 102000009027 Albumins Human genes 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 102000016942 Elastin Human genes 0.000 description 3
- 108010014258 Elastin Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 208000030961 allergic reaction Diseases 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 229940098773 bovine serum albumin Drugs 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 229920002549 elastin Polymers 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 229960001340 histamine Drugs 0.000 description 3
- 229920002674 hyaluronan Polymers 0.000 description 3
- 229960003160 hyaluronic acid Drugs 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 238000007619 statistical method Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 206010015150 Erythema Diseases 0.000 description 2
- 102000003960 Ligases Human genes 0.000 description 2
- 108090000364 Ligases Proteins 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 240000000220 Panda oleosa Species 0.000 description 2
- 235000016496 Panda oleosa Nutrition 0.000 description 2
- 208000003251 Pruritus Diseases 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000000692 Student's t-test Methods 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 230000003266 anti-allergic effect Effects 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 238000002306 biochemical method Methods 0.000 description 2
- 239000012496 blank sample Substances 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 210000004207 dermis Anatomy 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000001742 protein purification Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 238000012353 t test Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- WMBWREPUVVBILR-WIYYLYMNSA-N (-)-Epigallocatechin-3-o-gallate Chemical compound O([C@@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=C(O)C=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-WIYYLYMNSA-N 0.000 description 1
- FKKAGFLIPSSCHT-UHFFFAOYSA-N 1-dodecoxydodecane;sulfuric acid Chemical compound OS(O)(=O)=O.CCCCCCCCCCCCOCCCCCCCCCCCC FKKAGFLIPSSCHT-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- WMBWREPUVVBILR-UHFFFAOYSA-N GCG Natural products C=1C(O)=C(O)C(O)=CC=1C1OC2=CC(O)=CC(O)=C2CC1OC(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-UHFFFAOYSA-N 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
- 208000002720 Malnutrition Diseases 0.000 description 1
- -1 N-succinyl-alanine-p-nitroaniline Chemical compound 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 208000012641 Pigmentation disease Diseases 0.000 description 1
- 206010039897 Sedation Diseases 0.000 description 1
- 206010040880 Skin irritation Diseases 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002385 Sodium hyaluronate Polymers 0.000 description 1
- 206010042674 Swelling Diseases 0.000 description 1
- 206010044625 Trichorrhexis Diseases 0.000 description 1
- 206010000496 acne Diseases 0.000 description 1
- 239000013566 allergen Substances 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 231100000321 erythema Toxicity 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000003648 hair appearance Effects 0.000 description 1
- 230000003806 hair structure Effects 0.000 description 1
- 230000003699 hair surface Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 230000007803 itching Effects 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 150000002617 leukotrienes Chemical class 0.000 description 1
- 230000001071 malnutrition Effects 0.000 description 1
- 235000000824 malnutrition Nutrition 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 208000015380 nutritional deficiency disease Diseases 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 238000013492 plasmid preparation Methods 0.000 description 1
- 206010036067 polydipsia Diseases 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000011506 response to oxidative stress Effects 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 231100000475 skin irritation Toxicity 0.000 description 1
- 230000036556 skin irritation Effects 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 229940010747 sodium hyaluronate Drugs 0.000 description 1
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000009864 tensile test Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- 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
- A61Q19/005—Preparations for sensitive 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q5/00—Preparations for care of the hair
- A61Q5/002—Preparations for repairing the hair, e.g. hair cure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q5/00—Preparations for care of the hair
- A61Q5/12—Preparations containing hair conditioners
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Dermatology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to the field of cosmetics, in particular to application of recombinant spider silk protein. The recombinant spider silk protein provided by the invention has the application of any one of the following: (1) Use in the preparation of a product for soothing skin, anti-skin allergies; (2) Use in the preparation of a product for anti-wrinkle and tightening skin; (3) Use in the preparation of a product for improving the toughness of hair strands; the amino acid sequence of the recombinant spider silk protein comprises any one of the following: a. has an amino acid sequence shown as SEQ ID NO. 1; b. an amino acid sequence with the homology of more than or equal to 90 percent with the amino acid sequence shown in SEQ ID NO. 1; the research of the invention shows that the recombinant spider silk protein has the functions of relieving skin, resisting skin allergy, resisting skin wrinkle, tightening skin and improving hair bundle toughness, and can be used for preparing skin care products, cosmetics, hair washing products and hair care products.
Description
Technical Field
The invention relates to the field of cosmetics, in particular to application of recombinant spider silk protein.
Background
The skin serves as a first defense barrier for the human body, and has a function of protecting the organs in the body from the temperature and humidity changes and the external environment such as ultraviolet rays, nuisance substances and the like. Due to factors such as age, environment, irregular lifestyle, etc., the skin will change due to internal and external factors such as occurrence of wrinkles, age relaxation, dullness, pigmentation, itching, allergy, etc.
Skin aging is mainly affected by both intrinsic and extrinsic factors. Endogenous aging of the skin is determined by genetic background factors of the individual. External aging is caused by external factors such as smoking, excessive drinking, malnutrition and long-term exposure to sunlight. Externally, the skin is the outermost layer of a living body, and plays a role in protection. Thus, cumulative damage caused by environmental factors is considered to be a major cause of skin aging. In particular, ultraviolet light is considered to be the most serious environmental factor. Under the stimulation of ultraviolet rays, the expression and activity of Matrix Metalloproteinases (MMPs) in the skin are continuously improved, and MMPs are enzymes capable of degrading ECM components such as collagen, elastin and the like. The ultraviolet rays can stimulate the expression of MMPs, accelerate the damage to the integrity of the dermis layer and generate wrinkles. In addition, oxidative stress and inflammatory reaction of epidermis caused by ultraviolet rays can induce keratinocytes to synthesize MMPs and secrete the MMPs into dermis to act on collagen and elastin, thereby aggravating skin aging degree. The conventional skin care products have single protection effect on users and have insufficient report on crease-resistant effect.
Skin allergy is an allergic reaction, after an allergen enters a body, the body is promoted to generate corresponding antibodies, antigen-antibody reaction is induced, inflammatory substances such as histamine, leukotriene and the like are released, and symptoms such as erythema, pimple, pruritus, redness, swelling, dry scraps, blisters and the like are caused. With the pollution of air, environment and food and the excessive use of cosmetics, the human body has poorer resistance to the stimulation of foreign substances, and the phenomena of skin irritation, allergy and the like are more frequent. Hyaluronidase is a participant of type I allergic reaction, has strong correlation with inflammation and allergy, and researches report that various medicines capable of releasing histamine by fat large cells can regulate the activity of the hyaluronidase, and some anti-allergic medicines have strong inhibition of the activity of the hyaluronidase, so that active ingredients with the effect of inhibiting the activity of the hyaluronidase can be used for resisting skin allergy.
The hair contains a large amount of proteins, and various chemical bonds exist between protein chains, so that the strength and the shape of the hair structure are maintained. The daily processes of combing, blowing and scalding hair often cause the chemical bonds of the bearing structure in the hair to be broken, reduced or even broken, so that the hair quality is damaged and the strength and toughness of the hair are obviously deteriorated. However, most of the current hair care products focus on repairing the hair surface, and it is still a very challenging task to toughen the hair.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide the application of the recombinant spider silk protein, wherein the recombinant spider silk protein has the effects of relieving skin and resisting skin allergy, can be used for preparing products for relieving skin and resisting skin allergy, has the effects of resisting skin wrinkle and tightening skin, can be used for preparing products for resisting skin wrinkle and tightening skin, and has the effect of improving the toughness of hair bundles, and can be used for preparing products for improving the toughness of hair bundles.
For this purpose, the invention provides the following technical scheme:
a recombinant spider silk protein having the use of any one of:
(1) Use in the preparation of a product for soothing skin, anti-skin allergies;
(2) Use in the preparation of a product for anti-wrinkle and tightening skin;
(3) Use in the preparation of a product for improving the toughness of hair strands;
the amino acid sequence of the recombinant spider silk protein has the amino acid sequence shown as SEQ ID NO. 1.
Optionally, the use in the manufacture of a product for inhibiting hyaluronidase activity to relieve skin, anti-skin allergy.
Alternatively, use in the manufacture of a product for inhibiting elastase activity to wrinkle resistant and tighten skin.
Optionally, in the manufacture of a product for enhancing the tensile load and the total work to break of a hair strand to improve the toughness of the hair strand.
Optionally, the product comprises a cosmetic, skin care, hair wash or hair care product.
Optionally, the product comprises a cosmetic additive, a skin care additive, a shampoo additive or a hair care additive.
Optionally, in the product, the concentration of the recombinant spider silk protein is more than 0% v/v and less than or equal to 100% v/v.
Alternatively, the concentration of the recombinant spidroin solution in the product may be 0.01% v/v, 0.1% v/v, 1% v/v, 5% v/v, 6.25%, 10% v/v, 12.5%, 20% v/v, 25% v/v, 30% v/v, 40% v/v, 50% v/v, 60% v/v, 70% v/v, 80% v/v, 90% v/v, or 100% v/v.
Alternatively, the concentration of the recombinant spider silk protein in the product is 1% v/v to 50% v/v.
A cosmetic for soothing skin, preventing skin allergy, reducing skin wrinkles and/or tightening skin comprises the recombinant spider silk protein and acceptable auxiliary materials; the concentration of the recombinant spider silk protein is more than 0% v/v and less than or equal to 100% v/v.
A skin care product for soothing skin, resisting skin allergy, reducing skin wrinkles and/or tightening skin comprises the recombinant spider silk protein and acceptable auxiliary materials; the concentration of the recombinant spider silk protein is more than 0% v/v and less than or equal to 100% v/v.
A shampoo or hair care product for improving hair bundle toughness comprises the recombinant spider silk protein and acceptable auxiliary materials; the concentration of the recombinant spider silk protein is more than 0% v/v and less than or equal to 100% v/v.
The cosmetic for relieving skin, resisting skin allergy, resisting skin wrinkle and/or tightening skin, the skin care product for relieving skin, resisting skin allergy, resisting skin wrinkle and/or tightening skin or the hair washing product or hair care product for improving the toughness of hair bundles, wherein the concentration of the recombinant spider silk protein is 0.01% v/v, 0.1% v/v, 1% v/v, 5% v/v, 6.25% v/v, 10% v/v, 12.5% v/v, 20% v/v, 25% v/v, 30% v/v, 40% v/v, 50% v/v, 60% v/v, 70% v/v, 80% v/v, 90% v/v or 100% v/v.
The technical scheme of the invention has the following advantages:
1. the recombinant spider silk protein provided by the invention has the application of any one of the following: (1) Use in the preparation of a product for soothing skin, anti-skin allergies; (2) Use in the preparation of a product for anti-wrinkle and tightening skin; (3) Use in the preparation of a product for improving the toughness of hair strands; the amino acid sequence of the recombinant spider silk protein comprises any one of the following: a. has an amino acid sequence shown as SEQ ID NO. 1; b. an amino acid sequence with the homology of more than or equal to 90 percent with the amino acid sequence shown in SEQ ID NO. 1; the research of the invention shows that the recombinant spider silk protein has the effects of relieving skin and resisting skin allergy, so that the recombinant spider silk protein can be used for preparing products for relieving skin and resisting skin allergy; the recombinant spider silk protein also has the effects of resisting skin wrinkle and tightening skin, so that the recombinant spider silk protein can be used for preparing products for resisting skin wrinkle and tightening skin; in addition, the recombinant spider silk protein has the function of improving the toughness of hair bundles, and can be used for preparing products for improving the toughness of hair bundles.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the results of the inhibition of hyaluronidase activity by recombinant spider silk proteins at concentration gradients in example 2 of the present invention;
FIG. 2 is a graph showing the result of the inhibition of elastase activity by recombinant spider silk protein at concentration gradient in example 3 of the present invention;
FIG. 3 is a tensile load comparison of the recombinant spidroin protein of example 4 of the invention after application to hair, with a control group;
FIG. 4 is a comparison of total work to break in tension after application of the recombinant spidroin protein to hair in example 4 of the invention, compared to a control;
in the above figures, ""means p<0.05；“/> "means p<0.01；“/> "means p<0.001。
Detailed Description
The following examples are provided for a better understanding of the present invention and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the invention, any product which is the same or similar to the present invention, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present invention.
The specific experimental procedures or conditions are not noted in the examples and may be followed by the operations or conditions of conventional experimental procedures described in the literature in this field. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.
EXAMPLE 1 preparation of recombinant spider silk proteins
1. Construction of recombinant bacteria for recombinant spider silk proteins
1.1, synthesizing a target sequence: the gene sequence of the recombinant spider silk protein is synthesized by a entrusting gene synthesis company, two ends of the gene sequence are connected with EcoRI/HindIII enzyme digestion site sequences (5 '-cgaattc-3',5 '-caagcttg-3') respectively, and the sequences are shown as SEQ ID NO. 2.
1.2, inserting the recombinant spider silk protein sequence into a vector plasmid pET28a by an enzyme digestion connection assembly method, wherein the detailed steps are as follows:
a. insert preparation:
i. the synthesized fragment was digested with EcoRI/HindIII, and the correct fraction was recovered by cleavage with the following cleavage reaction system:
TABLE 1 configuration of cleavage reaction System (50. Mu.L)
Note that: the DNA fragment was subjected to 500 ng of PCR product.
ii. Enzyme digestion reaction: after the enzyme digestion system in the PCR tube is evenly mixed, the PCR tube is put into a PCR instrument for enzyme digestion at 37 ℃ (optimal reaction temperature) of 1h.
iii, fragment recovery procedure was performed at TIANgel Midi Purification Kit (commercially available).
iv, detecting the concentration of the recovered fragments in an enzyme-labeled instrument.
b. Vector plasmid preparation:
i. after 1. Mu.g of pET28a plasmid was digested with EcoRI/HindIII (the digestion reaction system and the digestion reaction conditions were the same as above), the correct fractions were recovered by digestion, and the vector backbone fragment concentration was examined.
c. Assembling a target plasmid:
i. the connection system is as follows:
TABLE 2 connection System
Note that: the T4 ligase/ligase buffer and BsaI used were both NEB brand.
ii. Connection conditions: mixing well after adding, and collecting the solution to the bottom of the tube by short-speed centrifugation to avoid the residue of the solution on the tube wall. The PCR tube was placed in a PCR apparatus and reacted at 16℃for 1h.
d. Transformation and screening
1) Conversion:
i. BL21 Chemically Competent Cell, a whole gold brand, was used as competent cells, and one was thawed on ice for 5min.
ii. 10 μl of the assembly reaction was added to 50 μl of competent cells, gently mixed, and ice-bathed for 30min.
iii, heat shock is carried out for 90s in a water bath kettle at 42 ℃.
iv immediately ice-bathed for 5min, then 500. Mu.l of LB medium was added, and shaking culture was performed at 37℃and 200rpm for 1h.
v, 200. Mu.l of the bacterial liquid was plated on LB solid plates containing 50mg/L kanamycin Kana, and cultured in an incubator at 37℃for 24 hours.
2) Screening:
i. from the transformation plate, 16 single colonies were picked, resuspended in 1.5ml sterile tube with 10 μl sterile water and numbered.
ii. Mu.l of the sample was subjected to PCR, and 1ml of LB liquid medium containing 50mg/L kanamycin was added to the corresponding sequence number, followed by shaking culture at 37℃and 200rpm for 24 hours.
TABLE 3 PCR verification primers
iii, PCR amplification System-50. Mu.l: the plasmid for synthesizing the target gene is used as a template, and the amplification system is shown in the following table 4:
TABLE 4 amplification System
iv, PCR amplification conditions are shown in Table 5 below:
TABLE 5 amplification conditions
Note that: denaturation-annealing-extension for 30 cycles.
v, gel electrophoresis verification: preparing 1% concentration agar gel; taking 5 μl of PCR product, and taking 5000bp DNA Maker as reference 140V for electrophoresis for 20min; the target band was correct at around 1881 bp.
vi, sequencing and verification: in a 1.5ml tube, 0.4ml of the correct bacterial liquid of the target fragment is taken, added with 0.4ml of 50% (v/v) glycerol, uniformly mixed, stored in a refrigerator at the temperature of minus 80 ℃, and the residual bacterial liquid is sent to Huada genes for verification of primers Ver-F and Ver-R for sequencing verification.
vii, strain preservation: and comparing the sequencing result, retaining a glycerol seed-retaining tube with correct sequencing, and constructing a plasmid marker pET28a-NT_2REP_CT.
1.3, the constructed strain is obtained by streaking a preserved glycerol tube on an LB solid medium plate containing 50mg/L kanamycin, and culturing the strain in a constant temperature incubator at 37 ℃ at 200 rpm. The monoclonal was picked up and shake cultivated overnight at 37℃in LB liquid medium containing 4-5 mL of 50mg/L kanamycin (Kana) at 200 rpm;
1.4 transfer to 400 mL shake flask fermentation at 1%, 200rpm, culture at 37℃2-3 h to OD 600 =0.6；
1.5, adding isopropyl thiogalactoside (IPTG) with the final concentration of 0.3-mM, inducing at 20 ℃ for 12-16 h,9000 g and centrifuging for 5-10 min to collect thalli.
2. Ultrasonic disruption of thallus
(1) Adding 20 mM Tris-HCl into the thalli to re-dissolve to obtain concentrated bacterial liquid, 40mL, and OD=20+.
(2) Ultrasonic 25 kHz, power 70-80%, ultrasonic 3 seconds each time, stopping for 6 seconds, and ultrasonic 30min altogether;
(3) The supernatant was centrifuged at 12000 rpm for 10-15 min.
3. Protein purification
(1) Repeatedly passing the ultrasonic supernatant through the filled nickel column for 2-3 times, and collecting the flow-through liquid;
(2) The column was washed 3-5 times (1-3 column volumes each) with 20 mM Tris-HCl until the dropped solution was clear;
(3) Washing the column material with 1-2 times of Pull Down buffer (commercially available) for at least 3 times, sequentially collecting, and recording as E1, E2 and E3;
(4) Washing the residual Pull down buffer with 5 column volumes of 20 mM Tris-HCl;
(5) If long-term storage is required after the column is used, 20 mM Tris-HCl is replaced by 20% alcohol for storage.
4. SDS-PAGE electrophoresis
(1) Preparing a 10% concentration separating gel according to the specification of an SDS-PAGE gel preparation kit (product number: P0012A);
(2) Running twice the protein glue: observing whether obvious target bands exist before and after the fermentation by first gel running (if no target bands exist, the collected thalli do not continue the protein purification step); the second run was performed by observing the eluted eluents E1, E2, E3, selecting a lot with distinct bands for dialysis, and simultaneously observing the flow-through, e.g., the flow-through still has a more distinct protein of interest, indicating that a second purification can be performed or an overnight incubation can be performed with an increased amount of column material for the protein supernatant of the next same run.
5. Dialysis of protein eluent and protein freeze-drying
(1) Pouring protein eluents E1 and E2 with obvious bands in the gel diagram into the same beaker, and filling into a dialysis bag;
(2) Dialyzing with pure water, immersing the dialysis bag in water, changing water for 2-3 h times, and changing water for 3-4 times;
(3) Quickly freezing the dialyzed protein liquid by liquid nitrogen, then placing the frozen protein liquid in a-80 refrigerator until the protein liquid is completely coagulated, and then placing the frozen protein liquid in a freeze dryer to freeze the protein;
(4) Weighing the freeze-dried protein, and storing the protein under a sealed low-temperature condition. The obtained recombinant spider silk protein is sequenced, the sequencing is correct, and the amino acid sequence is shown as SEQ ID NO. 1.
EXAMPLE 2 skin soothing and anti-skin sensitivity efficacy of recombinant spider silk proteins
It is known that hyaluronidase is a participant in type I allergic reaction, and has strong correlation with inflammation and allergy, and various medicines for releasing histamine by fat large cells are reported to be capable of regulating the activity of the hyaluronidase, and some anti-allergic medicines have strong inhibition of the activity of the hyaluronidase, so that active ingredients with the effect of inhibiting the activity of the hyaluronidase can be used for resisting skin allergy. Thus, the present example demonstrates that recombinant spidroin has the effect of soothing skin sensitivity by measuring the effect of the recombinant spidroin on inhibiting hyaluronidase activity.
(1) Preparing recombinant spider silk protein solution. Dissolving the recombinant spider silk protein prepared in the example 1 in water, preparing recombinant spider silk proteins with concentration gradients of 6.25% v/v, 12.5% v/v, 25% v/v, 50% v/v and 100% v/v (note: 100% of spider silk proteins are strains expressing the spider silk proteins through fermentation, extracting purified spider silk proteins from thalli to obtain a spider silk protein solution with the concentration of 1.1mg/ml, namely 100% (v/v), diluting the spider silk protein solution with 100% (v/v) samples, and refrigerating the spider silk protein solution with other concentrations of 100% (v/v) samples at 2-8 ℃ for later use as a test sample;
(2) 100. Mu.L of hyaluronidase (Sigma, 500U/mL), 100. Mu.L of PBS buffer (Gibco, 100 mmol/L, pH=7, 37 ℃), 100. Mu.L of NaCl (50 mmol/L), 100. Mu.L of BSA (0.01%) were mixed with 50. Mu.L of the test sample as a medium and incubated in a shaker (Henry, THZ-25) at 37℃for 10 min. Wherein the experimental group is concentration gradient recombinant spider silk protein solution in (1), the test sample of the blank control is PBS (pH value 7.0), and the test sample of the positive control is ascorbic acid (Sigma) (concentration is 2% w/v);
(3) Sodium hyaluronate (Hua Xi organism) was dissolved in 300 mmol/L phosphate solution (ph=5.35) to give 0.03% w/v hyaluronic acid solution, 100 μl hyaluronic acid solution was added to (2) as the start of the reaction, and incubated at 37 ℃ for 45 min;
(4) Dissolving 0.1% Bovine Serum Albumin (BSA) in 24 mmol/L sodium acetate (Guozhen) and 79mmol/L acetic acid (Guozhen, pH=3.75) to obtain acid albumin solutions, adding the acid albumin solutions to each group of solutions treated in the step (3), and precipitating the non-hydrolyzed hyaluronic acid with 1.0 mL acid albumin solution;
(5) After allowing the precipitated mixture to stand at room temperature for 10 min, detecting the absorbance of the mixture at 600 nm by using a microplate reader (Tecan, spark);
(6) All reagents should be freshly prepared prior to the enzymatic reaction. PBS group was used as a Blank (BC) and ascorbic acid (concentration 2% w/v) was used as a Positive Control (PC). The percentage calculation formula of the inhibition rate is as follows:
inhibition (%) = (absorbance of sample group/absorbance of blank group) ×100.
Referring to Table 6 and FIG. 1, in the experiment of inhibition of hyaluronidase activity (biochemical method), compared with the blank control group, the recombinant spider silk protein of the sample shows remarkable inhibition effect (p <0.05 or p < 0.01) on the hyaluronidase activity at the concentration of 6.25% -100% (V/V), and has the effects of relieving skin and resisting skin allergy.
TABLE 6 results of inhibition rate of hyaluronidase activity and table of the results
Remarks: when the statistical analysis is carried out by the t-test method, the significance of the sample group and the PC group is compared with the BC groupIt is indicated that p-value <0.05 is indicated as +.>P-value <0.01 is expressed as +.> . The concentration of the spidroin in the 100% (v/v) recombinant spidroin solution was 1.1. 1.1 mg/ml.
Example 3 recombinant spider silk proteins have anti-wrinkle and skin tightening effects
It is well known that elastase is the most important enzyme in the matrix metalloproteinase family, mainly secreted by fibroblasts, and is able to degrade elastin in the skin leading to skin aging. By measuring the effect of the recombinant spider silk protein on inhibiting the elastase activity, the recombinant spider silk protein can be clearly confirmed to have the effects of resisting skin wrinkles and tightening skin.
(1) Preparing recombinant spider silk protein solution. Dissolving the recombinant spider silk protein prepared in the example 1 in water, preparing the recombinant spider silk proteins with concentration gradients of 6.25% v/v, 12.5% v/v, 25% v/v, 50% v/v and 100% v/v (note: 100% spider silk protein is a strain expressing spider silk proteins through fermentation, extracting and purifying spider silk proteins from thalli to obtain a spider silk protein solution with the concentration of 1.1mg/ml, namely 100% (v/v), diluting the spider silk protein solution with 100% (v/v) samples, and refrigerating the spider silk solution with other concentrations of 100% (v/v) samples at 2-8 ℃ for later use as a test sample;
(2) Preparing a reaction solution and a method:
the blank samples were: tris-HCl buffer 140. Mu.L;
the blank control was: 130. Mu.L of Tris-HCl buffer+elastase (1.1. 1.1U/mL) 10. Mu.L;
the sample controls were: tris-HCl buffer 110. Mu.L+recombinant spider silk protein sample 30. Mu.L;
the samples are: 100. Mu.L of Tris-HCl buffer+elastase (1.1. 1.1U/mL) 10. Mu.L of recombinant spidroin sample 30. Mu.L;
positive Control (PC) was: tris-HCl buffer 100. Mu.L+elastase (1.1. 1.1U/mL) 10. Mu.L+EGCG (1 mg/mL) 30. Mu.L.
After the preparation of the above reaction solutions, the mixture was incubated at 25℃for 20min in a shaker, and then 40. Mu.L of AAAPVN (0.25 mg/mL) was added to each reaction solution in a unified manner, followed by incubation at 25℃for 20min in a shaker. The microplate reader measures the absorbance value at 410 nm. The calculation formula is as follows: inhibition ratio (%) = (1- (sample control-sample)/(blank control-blank sample)) ×100; AAAPVN is N-succinyl-alanine-p-nitroaniline.
Referring to Table 7 and FIG. 2, in the elastase activity inhibition (biochemical method) experiment, the recombinant spider silk protein sample showed significant inhibition (p <0.05 or p < 0.01) of elastase activity at a concentration of 6.25% -100% (V/V) compared with the control group, and can be used as one of the evidences for verifying the anti-wrinkle tightening effect.
TABLE 7 results summary of elastase Activity inhibition rate
Remarks: when the statistical analysis is carried out by the t-test method, the significance of the sample group and the PC group is compared with the BC groupIt is indicated that p-value <0.05 is indicated as +.>P-value <0.01 is expressed as +.> . The concentration of the spidroin in the 100% (v/v) recombinant spidroin solution was 1.1. 1.1 mg/ml.
Example 4 recombinant spider silk proteins have the effect of improving the toughness of hair strands
The embodiment examines that the recombinant spider silk protein has the effect of improving the toughness of hair bundles, and the method comprises the following steps:
(1) Obtaining an in-vitro hair bundle of a real person, and the specification is as follows: the length is 15-17 cm, the width is 1.5cm, and the weight is 2.5g;
(2) 2 hair bundles are randomly divided into 2 groups, 1 each of a control group and a sample group, and 1mL of 10% w/v SLES (sodium dodecyl ether sulfate) solution (basic shampoo) is used for basic cleaning before testing;
(3) According to the product usage instruction, the sample group hair bundle is uniformly smeared on the hair bundle by about 0.5g of 100% (v/v) recombined spider silk protein liquid (the mass ratio of the sample to the hair bundle is 1:5), stays for 1 minute, and is cleaned. The control group hair tresses did not use any product and the rest of the treatments were identical to the sample group. Suspending the hair bundle at 26+ -2deg.C, drying in constant temperature and humidity chamber with relative humidity of 60+ -10%, and balancing overnight; 100% (v/v) of a solution of recombinant spidroin at a spidroin concentration of 1.1 mg/ml;
(4) And respectively selecting 30 hairs from the hair bundles of the sample group and the control group for preparing samples, and using a hair multifunctional test system-Single hair tensile test fitting (Single fiber) for testing the tensile mechanical properties of the hairs, wherein the test indexes are the tensile load and the total tensile fracture work.
Referring to table 8, fig. 3 and 4, the tensile load and the total work at tensile break are shown for the control group and the sample group, respectively. It can be seen that the tensile load and the total work at tensile break result are both increased in the sample group compared to the control group. The SPSS software is used for carrying out statistical analysis on the hair tensile mechanical properties of the sample group and the control group, the test level alpha=0.05, and the tensile load and the total tensile breaking work of the sample group are obviously different from those of the control group (the tensile load p is less than 0.001, and the total tensile breaking work p is less than 0.001), so that the test sample has the effect of improving the strength and toughness of the hair bundle. In summary, the test specimens have an anti-breakage (improving the strength and toughness of the hair) effect.
Table 8, tensile load and tensile total work at break comparison
Application example 1
The application example provides a skin care product or cosmetic for relieving allergy and/or wrinkle and tightening skin, which contains the recombinant spider silk protein solution prepared in the example 1, and the balance of deionized water or conventional auxiliary materials; the concentration of the recombinant spidroin solution is > 0% v/v and less than or equal to 100% v/v, further, the concentration of the recombinant spidroin solution may be 0.01% v/v, 0.1% v/v, 1% v/v, 5% v/v, 6.25% v/v, 10% v/v, 12.5% v/v, 20% v/v, 25% v/v, 30% v/v, 40% v/v, 50% v/v, 60% v/v, 70% v/v, 80% v/v, 90% v/v, or 100% v/v.100% (v/v) of the recombinant spidroin solution was 1.1mg/ml of spidroin.
Application example 2
The application example provides a hair washing product or hair care product for improving the strength and toughness of hair bundles and preventing hair breakage, which comprises the recombinant spider silk protein solution prepared in the example 1, and deionized water or conventional auxiliary materials in balance; the concentration of the recombinant spidroin solution is > 0% v/v and less than or equal to 100% v/v, further, the concentration of the recombinant spidroin solution may be 0.01% v/v, 0.1% v/v, 1% v/v, 5% v/v, 6.25% v/v, 10% v/v, 12.5% v/v, 20% v/v, 25% v/v, 30% v/v, 40% v/v, 50% v/v, 60% v/v, 70% v/v, 80% v/v, 90% v/v, or 100% v/v.100% (v/v) of the recombinant spidroin solution was 1.1mg/ml of spidroin.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. And obvious variations or modifications thereof are contemplated as falling within the scope of the present invention.
Claims (10)
1. A recombinant spider silk protein having the use of any one of:
(1) Use in the preparation of a product for soothing skin, anti-skin allergies;
(2) Use in the preparation of a product for anti-wrinkle and tightening skin;
(3) Use in the preparation of a product for improving the toughness of hair strands;
the amino acid sequence of the recombinant spider silk protein has the amino acid sequence shown as SEQ ID NO. 1.
2. Use according to claim 1, characterized in that it is used for the preparation of a product that inhibits hyaluronidase activity to relieve skin, anti-skin allergies.
3. Use according to claim 1, characterized in that it is used for the preparation of a product for inhibiting elastase activity to wrinkle-resistant, skin-tightening the skin.
4. Use according to claim 1, characterized in that it is used in the preparation of products for enhancing the tensile load and the total work of tensile break of the hair strand to improve the toughness of the hair strand.
5. The use according to any one of claims 1 to 4, wherein the product comprises a cosmetic, skin care, hair wash or hair care product.
6. The use according to claim 5, wherein the product comprises a cosmetic additive, a skin care additive, a shampoo additive or a hair care additive.
7. The use according to claim 5, wherein the concentration of the recombinant spider silk protein in the product is 6.25% v/v, 12.5% v/v, 25% v/v, 50% v/v or 100% v/v.
8. A cosmetic for soothing skin, anti-skin allergy, anti-skin wrinkling and/or tightening skin, characterized by comprising recombinant spider silk proteins and acceptable auxiliary materials; the concentration of the recombinant spider silk protein is 6.25% v/v, 12.5% v/v, 25% v/v, 50% v/v or 100% v/v; the amino acid sequence of the recombinant spider silk protein has the amino acid sequence shown as SEQ ID NO. 1.
9. A skin care product for soothing skin, anti-skin allergy, anti-skin wrinkling and/or tightening skin, characterized by comprising recombinant spider silk proteins and acceptable auxiliary materials; the concentration of the recombinant spider silk protein is 6.25% v/v, 12.5% v/v, 25% v/v, 50% v/v or 100% v/v; the amino acid sequence of the recombinant spider silk protein has the amino acid sequence shown as SEQ ID NO. 1.
10. A hair washing or caring product for improving the toughness of hair bundles is characterized by comprising recombinant spider silk protein and acceptable auxiliary materials; the concentration of the recombinant spider silk protein is 6.25% v/v, 12.5% v/v, 25% v/v, 50% v/v or 100% v/v; the amino acid sequence of the recombinant spider silk protein has the amino acid sequence shown as SEQ ID NO. 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311669459.6A CN117357427A (en) | 2023-12-07 | 2023-12-07 | Application of recombinant spider silk protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311669459.6A CN117357427A (en) | 2023-12-07 | 2023-12-07 | Application of recombinant spider silk protein |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117357427A true CN117357427A (en) | 2024-01-09 |
Family
ID=89396917
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311669459.6A Pending CN117357427A (en) | 2023-12-07 | 2023-12-07 | Application of recombinant spider silk protein |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117357427A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103126918A (en) * | 2013-02-18 | 2013-06-05 | 崇州市地龙海龙生物制品开发研究所 | Preparation method for anti-wrinkle cream |
| CN103800273A (en) * | 2012-11-12 | 2014-05-21 | 烟台大洋制药有限公司 | Whitening and moistening cosmetics |
| KR101508614B1 (en) * | 2014-08-25 | 2015-04-08 | 한솔바이오 주식회사 | Anti-aging cosmetic composition comprising the poison complex of spider silk protein, leech extracts, bee venom extracts and snake venom peptide |
| CN109689680A (en) * | 2016-07-01 | 2019-04-26 | 思百博技术股份公司 | It is engineered spider silk fibroin and application thereof |
| WO2021083616A1 (en) * | 2019-10-31 | 2021-05-06 | Givaudan Sa | Hair care composition |
| KR102346006B1 (en) * | 2021-05-21 | 2022-01-03 | (주)파이온텍 | Pentapeptide derived from spider web and composition for anti-aging containing the same |
-
2023
- 2023-12-07 CN CN202311669459.6A patent/CN117357427A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103800273A (en) * | 2012-11-12 | 2014-05-21 | 烟台大洋制药有限公司 | Whitening and moistening cosmetics |
| CN103126918A (en) * | 2013-02-18 | 2013-06-05 | 崇州市地龙海龙生物制品开发研究所 | Preparation method for anti-wrinkle cream |
| KR101508614B1 (en) * | 2014-08-25 | 2015-04-08 | 한솔바이오 주식회사 | Anti-aging cosmetic composition comprising the poison complex of spider silk protein, leech extracts, bee venom extracts and snake venom peptide |
| CN109689680A (en) * | 2016-07-01 | 2019-04-26 | 思百博技术股份公司 | It is engineered spider silk fibroin and application thereof |
| WO2021083616A1 (en) * | 2019-10-31 | 2021-05-06 | Givaudan Sa | Hair care composition |
| KR102346006B1 (en) * | 2021-05-21 | 2022-01-03 | (주)파이온텍 | Pentapeptide derived from spider web and composition for anti-aging containing the same |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lancefield et al. | Preparation and properties of type-specific M antigen isolated from a group A, type 1 hemolytic streptococcus | |
| Chaim et al. | Brown spider dermonecrotic toxin directly induces nephrotoxicity | |
| US6077518A (en) | Cloning of mite allergens | |
| Lavaud et al. | Allergy to latex, avocado pear, and banana: evidence for a 30 kd antigen in immunoblotting | |
| da Silveira et al. | Molecular cloning and functional characterization of two isoforms of dermonecrotic toxin from Loxosceles intermedia (brown spider) venom gland | |
| CN109593127B (en) | Gene recombinant collagen peptide MJ L GG-34 and preparation method and application thereof | |
| da Silveira et al. | Two novel dermonecrotic toxins LiRecDT4 and LiRecDT5 from brown spider (Loxosceles intermedia) venom: from cloning to functional characterization | |
| CN103102407A (en) | Genetic recombinant human-like collagen | |
| CN114106202B (en) | Conus toxin and collagen fusion protein and preparation method and application thereof | |
| CN106093435B (en) | The application of Echinococcus granulosus Glutaredoxin 1 | |
| CN111840530B (en) | Preparation of recombinant polypeptide vaccine VNQS of Eimeria tenella and application method of recombinant polypeptide vaccine VNQS in resisting chicken coccidiosis | |
| CN101775072A (en) | Glial cell line-Derived Neurotrophic Factor (GDNF) fusing penetrating peptides | |
| CN117357427A (en) | Application of recombinant spider silk protein | |
| CN116970071B (en) | Recombinant elastin with anti-aging activity and preparation method and application thereof | |
| CN109939227A (en) | A kind of ragweed pollen allergen extract, its immersion liquid and preparation method thereof | |
| JP7024004B2 (en) | Screening method for preventive / ameliorating agents for keratin plug formation | |
| Bungy et al. | Mapping of T cell epitopes of the major fraction of rye grass using peripheral blood mononuclear cells from atopics and non‐atopics. II. Isoallergen clone 5A of Loliuum perenne group I (Lol p I) | |
| CN112876569B (en) | rhTSG6-FN III1-C fusion protein, application thereof in skin care composition and preparation method thereof | |
| Parkkinen et al. | Homology of a bovine allergen and the oligomycin sensitivity–conferring protein of the mitochondrial adenosine triphosphate synthase complex | |
| CN109265553B (en) | Fusion protein of cytoglobin and sipunculus nudus plasmin | |
| CN108103048B (en) | Low-temperature matrix metalloproteinase and coding gene and application thereof | |
| CN110064052B (en) | England phoenix tree pollen allergen extract, its extract and preparation method thereof | |
| CN101570760A (en) | Recombinant mouse beta-alexin 3 polypeptide, preparation and use thereof | |
| CN112423769A (en) | Extract for treating sensitive skin and dermatological composition containing the same | |
| CN117247442B (en) | Frog genus anion active peptide and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |