CN111662377A - Preparation method of mixed material for rapidly removing wrinkles and rejuvenating skin - Google Patents
Preparation method of mixed material for rapidly removing wrinkles and rejuvenating skin Download PDFInfo
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- CN111662377A CN111662377A CN202010685425.6A CN202010685425A CN111662377A CN 111662377 A CN111662377 A CN 111662377A CN 202010685425 A CN202010685425 A CN 202010685425A CN 111662377 A CN111662377 A CN 111662377A
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- bacterium
- mixed material
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- genetic engineering
- dialysis
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- 239000000463 material Substances 0.000 title claims abstract description 13
- 230000037303 wrinkles Effects 0.000 title claims abstract description 8
- 230000003716 rejuvenation Effects 0.000 title claims abstract description 6
- 238000002360 preparation method Methods 0.000 title abstract description 6
- 241000894006 Bacteria Species 0.000 claims abstract description 21
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 9
- 238000000502 dialysis Methods 0.000 claims abstract description 8
- 238000010353 genetic engineering Methods 0.000 claims abstract description 8
- 239000013612 plasmid Substances 0.000 claims abstract description 8
- 238000000746 purification Methods 0.000 claims abstract description 6
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 5
- 108060003393 Granulin Proteins 0.000 claims abstract description 4
- 239000001110 calcium chloride Substances 0.000 claims abstract description 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims abstract description 4
- 238000010367 cloning Methods 0.000 claims abstract description 4
- 238000012258 culturing Methods 0.000 claims abstract description 4
- 238000000855 fermentation Methods 0.000 claims abstract description 4
- 230000004151 fermentation Effects 0.000 claims abstract description 4
- 239000001963 growth medium Substances 0.000 claims abstract description 4
- 239000012535 impurity Substances 0.000 claims abstract description 4
- 238000001742 protein purification Methods 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 102000012422 Collagen Type I Human genes 0.000 claims description 3
- 108010022452 Collagen Type I Proteins 0.000 claims description 3
- 102000004266 Collagen Type IV Human genes 0.000 claims description 3
- 108010042086 Collagen Type IV Proteins 0.000 claims description 3
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 claims description 3
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 claims description 3
- 102000016359 Fibronectins Human genes 0.000 claims description 3
- 108010067306 Fibronectins Proteins 0.000 claims description 3
- 229940096422 collagen type i Drugs 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 abstract description 3
- 241000588724 Escherichia coli Species 0.000 abstract description 3
- 235000011148 calcium chloride Nutrition 0.000 abstract description 3
- 230000002068 genetic effect Effects 0.000 abstract description 2
- 238000011426 transformation method Methods 0.000 abstract 1
- 230000001131 transforming effect Effects 0.000 abstract 1
- 102000008186 Collagen Human genes 0.000 description 3
- 108010035532 Collagen Proteins 0.000 description 3
- 229920001436 collagen Polymers 0.000 description 3
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- 206010052428 Wound Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 230000000399 orthopedic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000017423 tissue regeneration Effects 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- 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
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Public Health (AREA)
- Biotechnology (AREA)
- Animal Behavior & Ethology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Gerontology & Geriatric Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dermatology (AREA)
- Plant Pathology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a preparation method of a mixed material for rapidly removing wrinkles and rejuvenating skin, which comprises the following steps: cloning and inserting the coding gene of the outer matrix protein into a plasmid pET-32a (+); transforming into competent cell Escherichia coli cell BL21(DE3) by CaCl2 transformation method; obtaining genetically engineered bacteria containing the recombinant plasmid; fermentation and thallus collection: selecting the positive genetic engineering bacterium clone obtained in the step S3, placing the positive genetic engineering bacterium clone in a culture medium for culturing until the bacterium reaches the target concentration, and then collecting the bacterium; purification and dialysis: and (4) removing impurities from the thallus collected in the step S4, performing protein purification operation, and performing protein dialysis operation to obtain the mixed material. The invention has the advantages of clear genetic background, quick propagation, low cost, high expression quantity, easy purification of expression products, good stability, strong pollution resistance and the like.
Description
Technical Field
The invention belongs to the field of recombinant protein expression, and particularly relates to a preparation method of a mixed material for rapidly removing wrinkles and enabling skin to be young.
Background
Collagen is widely distributed in the body of multicellular animals. Collagen has been thought to function only as a supporting tissue protein. In recent years, research on collagen has been increasing, and due to its excellent biological properties and weak antigenicity, it can be used in the medical and health fields of burns, wounds, beauty, orthopedics, tissue repair, wound hemostasis, etc. after being specially treated, and this research has been successful. Therefore, it is necessary to research and develop the synthesized recombinant protein using the related art of genetically engineered bacteria.
Disclosure of Invention
The invention mainly aims to overcome the defects of the prior art and provide a preparation method of a mixed material for rapidly removing wrinkles and rejuvenating skin.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing a mixed material for rapidly removing wrinkles to rejuvenate skin, the method comprising the steps of:
s1, cloning and inserting the coding gene of the extracellular matrix protein into a plasmid pET-32a (+);
s2 was transformed into competent cell E.coli cell BL21(DE3) by CaCl2 transformation;
s3, obtaining a genetically engineered bacterium containing the recombinant plasmid;
s4 fermentation and thallus collection: selecting the positive genetic engineering bacterium clone obtained in the step S3, placing the positive genetic engineering bacterium clone in a culture medium for culturing until the bacterium reaches the target concentration, and then collecting the bacterium;
s5 purification and dialysis: and (4) removing impurities from the thallus collected in the step S4, performing protein purification operation, and performing protein dialysis operation to obtain the mixed material.
Preferably, the matrix proteins include collagen type I, collagen type IV, fibronectin.
The invention also provides application of the prepared mixed material in the field of skin care products.
The invention has the advantages of clear genetic background, quick propagation, low cost, high expression quantity, easy purification of expression products, good stability, strong pollution resistance and the like.
Detailed Description
The present invention will be further described below, and it should be noted that the following examples are provided to illustrate the detailed embodiments and specific procedures based on the technical solution, but the scope of the present invention is not limited to the examples.
The invention relates to a preparation method of a mixed material for rapidly removing wrinkles and rejuvenating skin, which comprises the following steps:
s1, cloning and inserting the coding gene of the extracellular matrix protein into a plasmid pET-32a (+);
s2 was transformed into competent cell E.coli cell BL21(DE3) by CaCl2 transformation;
s3, obtaining a genetically engineered bacterium containing the recombinant plasmid;
s4 fermentation and thallus collection: selecting the positive genetic engineering bacterium clone obtained in the step S3, placing the positive genetic engineering bacterium clone in a culture medium for culturing until the bacterium reaches the target concentration, and then collecting the bacterium;
s5 purification and dialysis: and (4) removing impurities from the thallus collected in the step S4, performing protein purification operation, and performing protein dialysis operation to obtain the mixed material.
Preferably, the matrix proteins include collagen type I, collagen type IV, fibronectin.
The invention also provides application of the prepared mixed material in the field of skin care products.
Various corresponding changes and modifications can be made by those skilled in the art based on the above technical solutions and concepts, and all such changes and modifications should be included in the protection scope of the present invention.
Claims (3)
1. A method for preparing a skin rejuvenating composition for rapid wrinkle removal, said method comprising the steps of:
s1, cloning and inserting the coding gene of the extracellular matrix protein into a plasmid pET-32a (+);
s2 passing through CaCl2Transformation into competent cells by transformationEnterobacter cells BL21(DE 3);
s3, obtaining a genetically engineered bacterium containing the recombinant plasmid;
s4 fermentation and thallus collection: selecting the positive genetic engineering bacterium clone obtained in the step S3, placing the positive genetic engineering bacterium clone in a culture medium for culturing until the bacterium reaches the target concentration, and then collecting the bacterium;
s5 purification and dialysis: and (4) removing impurities from the thallus collected in the step S4, performing protein purification operation, and performing protein dialysis operation to obtain the mixed material.
2. The method of claim 1, wherein the matrix proteins include collagen type I, collagen type IV, and fibronectin.
3. Use of the mixture prepared according to claim 1 in the field of skin care products.
Priority Applications (1)
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CN202010685425.6A CN111662377A (en) | 2020-07-16 | 2020-07-16 | Preparation method of mixed material for rapidly removing wrinkles and rejuvenating skin |
Applications Claiming Priority (1)
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CN202010685425.6A CN111662377A (en) | 2020-07-16 | 2020-07-16 | Preparation method of mixed material for rapidly removing wrinkles and rejuvenating skin |
Publications (1)
Publication Number | Publication Date |
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CN111662377A true CN111662377A (en) | 2020-09-15 |
Family
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Family Applications (1)
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CN202010685425.6A Pending CN111662377A (en) | 2020-07-16 | 2020-07-16 | Preparation method of mixed material for rapidly removing wrinkles and rejuvenating skin |
Country Status (1)
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CN (1) | CN111662377A (en) |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0330396B1 (en) * | 1988-02-26 | 1995-06-21 | Biogen, Inc. | DNA sequences, recombinant DNA molecules and processes for producing lipocortins III, IV, V & VI |
US20080014613A1 (en) * | 2003-04-23 | 2008-01-17 | Magnus Hook | Prokaryotic collagen-like proteins and uses thereof |
CN102061296A (en) * | 2010-08-20 | 2011-05-18 | 吉林农业大学 | Preparation method of water-soluble human collagen VI polypeptide |
CN103122027A (en) * | 2012-11-26 | 2013-05-29 | 杨霞 | Recombinant human collagen and production method thereof |
CN110003324A (en) * | 2019-03-19 | 2019-07-12 | 江苏悦智生物医药有限公司 | Recombination human source collagen and its application |
CN110194795A (en) * | 2019-06-13 | 2019-09-03 | 郭伟 | A kind of recombination human source collagen and its application |
CN110903383A (en) * | 2019-11-21 | 2020-03-24 | 郭伟 | Recombinant human type I collagen, encoding gene, engineering bacterium and application thereof |
CN111087464A (en) * | 2019-12-28 | 2020-05-01 | 河北纳科生物科技有限公司 | Recombinant human III-type collagen with functional structure and expression method thereof |
-
2020
- 2020-07-16 CN CN202010685425.6A patent/CN111662377A/en active Pending
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0330396B1 (en) * | 1988-02-26 | 1995-06-21 | Biogen, Inc. | DNA sequences, recombinant DNA molecules and processes for producing lipocortins III, IV, V & VI |
US20080014613A1 (en) * | 2003-04-23 | 2008-01-17 | Magnus Hook | Prokaryotic collagen-like proteins and uses thereof |
CN102061296A (en) * | 2010-08-20 | 2011-05-18 | 吉林农业大学 | Preparation method of water-soluble human collagen VI polypeptide |
CN103122027A (en) * | 2012-11-26 | 2013-05-29 | 杨霞 | Recombinant human collagen and production method thereof |
CN110003324A (en) * | 2019-03-19 | 2019-07-12 | 江苏悦智生物医药有限公司 | Recombination human source collagen and its application |
CN110194795A (en) * | 2019-06-13 | 2019-09-03 | 郭伟 | A kind of recombination human source collagen and its application |
CN110903383A (en) * | 2019-11-21 | 2020-03-24 | 郭伟 | Recombinant human type I collagen, encoding gene, engineering bacterium and application thereof |
CN111087464A (en) * | 2019-12-28 | 2020-05-01 | 河北纳科生物科技有限公司 | Recombinant human III-type collagen with functional structure and expression method thereof |
Non-Patent Citations (8)
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JINGJING SHI等: "Hydroxylation of Human Type III Collagen Alpha Chain by Recombinant Coexpression with a Viral Prolyl 4-Hydroxylase in Escherichia coli", 《THE PROTEIN JOURNAL》 * |
NAVIT OGEN‐SHTERN等: "Increased pro‐collagen 1, elastin, and TGF‐β1 expression by copper ions in an ex‐vivo human skin model", 《JOURNAL OF COSMETIC DERMATOLOGY》 * |
YUNPING TANG等: "Efficient Production of Hydroxylated Human-Like Collagen Via the Co-Expression of Three Key Genes in Escherichia coli Origami (DE3)", 《APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY》 * |
吴铭等: "胶原蛋白在化妆品中的应用及研究进展", 《日用化学品科学》 * |
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Application publication date: 20200915 |