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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 polypeptides, so as to improve transdermal properties, Low cost, easy preparation and low effect

Active Publication Date: 2021-12-17
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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  • Recombinant protein and its construction method and use
  • Recombinant protein and its construction method and use
  • Recombinant protein and its construction method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The research and development of embodiment 1 transdermal fibronectin mutant

[0041] 1. Mutant research and development ideas:

[0042] 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.

[0043] In the present invention, the transdermal peptide and fibronectin are recombined, the sequence of the recombinant transdermal fibronectin is shown in SEQ ID No.6, and then according to the three-dimensional structural characteristics of the recombinant transdermal fibronectin, amino acid point mutations are carried out on the surface of the recombinant transdermal fibronectin, It can significantly improve the stability of recombinant transdermal fibronectin. The selection of mutation points should not ...

Embodiment 2

[0050] Example 2 Construction of Transdermal Fibronectin Mutant Recombinant Protein Particles

[0051] In the embodiment of the present invention, the amino acid sequence of the recombinant transdermal fibronectin mutant (A-7) is shown in SEQ ID No.1, and the gene sequence encoding SEQ ID No.1 is shown in SEQ ID No.2.

[0052] Build method:

[0053] (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 in SEQ ID No.2;

[0054] (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;

[0055] (3) Insert the target fragment of the recombinant protein into the restriction site of the vector to obtain the expression plasmid encoding the above recombinant protein. The structure of the vecto...

Embodiment 3

[0056] Example 3 Construction of Transdermal Fibronectin Mutant-Elastase Inhibiting Peptide Recombinant Protein Particles

[0057] 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 connected to the elastase inhibitory peptide through a connecting peptide, the recombinant protein The amino acid sequence of is shown in SEQ ID No.4.

[0058] Build method:

[0059] (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 fibronectin mutant-elastase inhibitory peptide recombinant protein is shown in SEQ ID No.5;

[0060] (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 respective...

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Abstract

The invention discloses a recombinant protein, which consists of transdermal fibronectin connected by connecting peptides to form recombinant transdermal fibronectin, and then N138Q, E174D, G230V and P292G site mutations are performed on the amino acid sequence of the recombinant transdermal fibronectin A recombinant transdermal fibronectin mutant is constructed, the amino acid sequence of the recombinant transdermal fibronectin mutant is shown in SEQ ID No.1, and the elastase inhibitory peptide is connected to the flexible end of the recombinant transdermal fibronectin mutant through a connecting peptide Build gets. The recombinant protein provided by the invention has better stability and guaranteed efficacy, and is easy to prepare and lower in cost, and greatly expands the application of fibronectin and elastase inhibitory peptide 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 王娟李宇涵
Owner MEIMU (BEIJING) TECH CO LTD