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Long-acting recombinant metallothionein expressed by saccharomyces cerevisiae and application of long-acting recombinant metallothionein in cosmetics

A metallothionein and Saccharomyces cerevisiae technology, applied in metallothionein, cosmetics, cosmetic preparations, etc., can solve the problem of unclear mechanism of action, and achieve the effects of detoxification, absorption promotion, and free radical scavenging.

Inactive Publication Date: 2021-07-30
WUHU YINGTE FEIER BIOLOGICAL PROD IND RES INST CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Studies have found that MT is related to various functions in organisms, mainly including scavenging free radicals, activating (inactivating) Zn regulatory proteins, etc., among which the most basic and main function is to combine with metal ions to release or reduce biological life. The toxic effects of heavy metals in the environment, however, so far, the mechanism of action of many functions of MT is still unclear

Method used

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  • Long-acting recombinant metallothionein expressed by saccharomyces cerevisiae and application of long-acting recombinant metallothionein in cosmetics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] This example provides a long-acting recombinant metallothionein expressed by Saccharomyces cerevisiae. The long-acting recombinant metallothionein has the advantages of scavenging free radicals, activating (inactivating) Zn regulatory proteins, preventing body aging, and degrading the toxic effects of heavy metals.

[0028] The nucleotide sequence of the long-acting recombinant metallothionein is shown in Seq ID NO.1. The amino acid sequence corresponding to the nucleotide sequence of the long-acting recombinant metallothionein is shown in Seq ID NO.2.

[0029] The nucleotide sequence of long-acting recombinant metallothionein is obtained from the original sequence of metallothionein after yeast codon optimization. The nucleotide sequence of the original sequence of metallothionein after yeast codon optimization includes Not I restriction site, Xba I restriction site, start codon, stop codon and 6×His tag sequence. The nucleotide sequence of the Not I restriction site ...

Embodiment 2

[0037] In order to verify the activity of the long-acting recombinant metallothionein described in Example 1, this example provides the activity detection of the long-acting recombinant metallothionein expressed by Saccharomyces cerevisiae.

[0038] The detection reagents used in the detection include: Hydroxyl radical scavenging activity detection kit (Beijing Legen Biotechnology Co., Ltd., article number TO1131, specification 100T). The detection principle is H 2 o 2 / Fe 2+ Hydroxyl radicals are generated by the Fenton reaction, and the Fe 2 + Oxidized to Fe 3+ , the generated sulfinic acid reacts with azo dyes to generate azo haze, and the absorbance at 420nm is measured with a microplate reader. The color depth is proportional to the generated hydroxyl radicals within a certain range, and can be calculated by analyzing the captured hydroxyl radicals Free radical scavenging rate or scavenging capacity.

[0039]The measuring operation of this detection comprises: first...

Embodiment 3

[0047] This example provides an application of Saccharomyces cerevisiae expressing long-acting recombinant metallothionein in cosmetics, which uses Saccharomyces cerevisiae as described in Example 1 to express long-acting recombinant metallothionein. The long-acting recombinant metallothionein has the advantages of scavenging free radicals, activating (inactivating) Zn regulatory proteins, preventing body aging, degrading toxic effects of heavy metals, and the like. And as described in Example 2, the long-acting recombinant metallothionein in Example 1 has a strong ability to scavenge free radicals and can effectively prevent aging of the body. It can be seen that the long-acting recombinant metallothionein described in Example 1 has broad application prospects in cosmetics.

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Abstract

The invention discloses long-acting recombinant metallothionein expressed by saccharomyces cerevisiae and an application of the long-acting recombinant metallothionein in cosmetics. The nucleotide sequence of the long-acting recombinant metallothionein is as shown in Seq ID NO. 1. The nucleotide sequence of the long-acting recombinant metallothionein is obtained by optimizing an original sequence of the metallothionein through a yeast codon. Compared with metallothionein, the long-acting recombinant metallothionein provided by the invention contains a 6*histidine tags, can realize metal ion affinity chromatography protein purification, and also has the advantages of degrading heavy metal toxicity, scavenging free radicals, activating (inactivating) Zn regulatory protein, preventing body aging and the like.

Description

technical field [0001] The present invention relates to the emerging field of biomedicine, in particular to a long-acting recombinant metallothionein expressed by Saccharomyces cerevisiae, an activity detection method of long-acting recombinant metallothionein, and a long-acting recombinant metallothionein expressed by Saccharomyces cerevisiae in cosmetics Applications. Background technique [0002] Metallothionein (MT) is a low-molecular-weight, cysteine-rich metal-binding protein. The common characteristics of MT are: low molecular weight (6-7kDa); high metal content (7-12 metal ions) ; High cysteine ​​content (20-30% Cys-X-Cys sequence), lack of aromatic amino acids, very little histidine content; composed of α domain and β domain. Animal MT can be divided into four subtypes: MT-1, MT-2, MT-3 and MT-4 according to the difference in isoelectric point and amino acid composition. MT-4 is expressed in the stratified squamous epithelial cells of oral epithelium, esophagus, n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C07K19/00A61K8/64A61Q19/08A61Q19/00G01N21/31
CPCA61K8/64A61Q19/00A61Q19/08C07K14/825C07K2319/21G01N21/31
Inventor 赵俊刘洁许高涛李媛媛杨欣怡夏兵兵张勇
Owner WUHU YINGTE FEIER BIOLOGICAL PROD IND RES INST CO LTD
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