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A kind of recombinant protein and its construction method and use

A technology of recombinant protein and fibronectin, which is applied in the field of recombinant protein, can solve the problems of poor transdermal performance of protein-polypeptide complexes, poor stability of fibronectin, and restrictions on the application of protein and polypeptide, so as to improve transdermal performance, The preparation is simple and low cost, and the effect of expanding application

Active Publication Date: 2021-11-23
MEIMU (BEIJING) TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the prior art, the above-mentioned small molecule polypeptide and fibronectin are generally compounded by physical mixing method in the cosmetic formula. The stability of fibronectin itself is not good, and the small molecule polypeptide is added again, and the challenge of stable performance is even greater, and the preparation cost also higher
At the same time, because there is a physical barrier on the surface of the skin, most of the macromolecular active ingredients are prevented from acting through the skin, because the transdermal performance of the protein-peptide complex is often poor, so that the originally highly active polypeptide cannot fully exert its due effect. effect, which further limits the application of protein peptides in cosmetics

Method used

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  • A kind of recombinant protein and its construction method and use
  • A kind of recombinant protein and its construction method and use
  • A kind of recombinant protein and its construction method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The research and development of embodiment 1 transdermal fibronectin mutant

[0039] 1. Mutant research and development ideas:

[0040] There are surfactants, thickeners, essences, polyols and other ingredients in cosmetic formulations, which will affect the stability of protein peptides in cosmetics. At the same time, cosmetics may encounter high temperature conditions during transportation and storage, which will accelerate the degradation of protein peptides.

[0041] According to the three-dimensional structural characteristics of the recombinant protein after recombining the transdermal peptide and fibronectin, the original sequence of the transdermal peptide fibronectin is shown in SEQ ID No.6, and amino acid point mutations are carried out on its surface, which can significantly improve the recombinant protein The stability of the protein, the mutation point selection effectively avoids the active site, and does not have any impact on the function of the recombi...

Embodiment 2

[0048] Example 2 Construction of Transdermal Fibronectin Mutant Recombinant Protein Particles

[0049] In the embodiment of the present invention, the amino acid sequence of the recombinant transdermal fibronectin mutant (A-5) is shown in SEQ ID No.1, wherein ACSPPHSKSHC is a transdermal peptide sequence, which can enhance the transdermal absorption performance of the protein, GGGGS For the link between the transdermal peptide and fibronectin.

[0050] Build method:

[0051] (1) According to the position design of the relevant sequence of the commercial vector pPICZaA, the enzyme cutting sites EcoR I and SalI were selected; the DNA sequence encoding the transdermal fibronectin mutant recombinant protein is shown as SEQ ID No.2;

[0052] (2) There is a restriction endonuclease site at the 5' and 3' ends of the synthetic recombinant protein DNA sequence, corresponding to EcoR I and SalI respectively;

[0053] (3) Insert the target fragment of the recombinant protein into the r...

Embodiment 3

[0054] Example 3 Construction of Transdermal Fibronectin Mutant-Elastase Inhibiting Peptide Recombinant Protein Particles

[0055] In the embodiment of the present invention, the transdermal fibronectin mutant-elastase inhibitory peptide recombinant protein is the flexible end of the recombinant transdermal fibronectin mutant prepared in Example 2, which is connected to the elastase inhibitory peptide through a connecting peptide. The amino acid sequence is shown in SEQ ID No.4.

[0056] Build method:

[0057] (1) According to the position design of the relevant sequence of the commercial vector pPICZaA, the enzyme cleavage sites EcoR I and SalI were selected; the DNA sequence encoding the transdermal peptide-fibronectin mutant-elastase inhibitory peptide recombinant protein is shown in SEQ ID No.5;

[0058] (2) There is a restriction endonuclease site at the 5' and 3' ends of the synthetic recombinant protein DNA sequence, corresponding to EcoR I and SalI respectively;

[0...

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Abstract

The invention discloses a recombinant protein, which consists of a transdermal peptide connected to fibronectin through a connecting peptide to form a recombinant transdermal fibronectin, and then carries out E174D and P292G site mutations on the amino acid sequence of the recombinant transdermal fibronectin to obtain a recombinant transdermal fibronectin. A fibronectin mutant, the amino acid sequence of the recombinant transdermal fibronectin mutant is shown in SEQ ID No.1, and the flexible end of the recombinant transdermal fibronectin mutant is connected to an elastase inhibitor by a connecting peptide Peptides are constructed. The recombinant protein provided by the invention has better stability and guaranteed efficacy, is easy to prepare and has low cost, and greatly expands the application of fibronectin and collagenase inhibitory peptides in cosmetics.

Description

technical field [0001] The invention relates to a recombinant protein, in particular to a highly stable recombinant protein used for cosmetic preparation or addition and its construction method. Background technique [0002] Fibronectin (also known as Fn) is a high molecular weight glycoprotein in the extracellular matrix. FN is produced by liver and vascular endothelial cells and widely exists in animal tissues and interstitial fluids. It is a macromolecular glycoprotein with the biological function of cell regeneration and repair. It has been used in clinical medicine, including the treatment of spinal cord. Sickness, burns, etc. Experience has proved that fibronectin has a repairing effect on sensitive skin, red blood streaks, acne, sun damage and other skin problems. [0003] Although fibronectin has excellent skin care effects, its thermal stability and chemical stability are not good. Poor thermal stability is mainly reflected in the precipitation, precipitation or ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/81A61K8/64A61Q19/00A61Q19/08A61P17/10
CPCC07K14/78C07K14/811C12N15/815A61K8/64A61Q19/004A61Q19/005A61Q19/08A61P17/10C07K2319/00
Inventor 李宇涵王娟
Owner MEIMU (BEIJING) TECH CO LTD
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