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
- Publication number
- 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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- China
- Prior art keywords
- bacterium
- mixed material
- purification
- genetic engineering
- dialysis
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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, cold insoluble globulin [CIG]
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- 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
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- 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
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- 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
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- 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
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)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| 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)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| 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 |
|---|---|
| CN111662377A true CN111662377A (en) | 2020-09-15 |
Family
ID=72392873
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| 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)
| Country | Link |
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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)
| Title |
|---|
| 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》 * |
| 唐云平等: "重组胶原蛋白制备及其应用研究进展", 《食品工业科技》 * |
| 李虹等: "《广东省医疗美容医师培训教材系列丛书 美容技术分册》", 30 November 2015, 人民军医出版社 * |
| 杨晶等: "重组类人Ⅰ型胶原蛋白肽在大肠杆菌中的表达纯化及功能鉴定", 《现代食品科技》 * |
| 王皓等: "类人胶原蛋白基因COL6A2的克隆及其在大肠杆菌中的表达", 《吉林农业大学学报》 * |
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Application publication date: 20200915 |