Halostachys caspica metallothionein HcMT compound and application thereof in sunscreen cream

A technology of metallothionein and salt ear wood, applied in the direction of metallothionein, hybrid peptide, fusion polypeptide, etc., can solve the problems of difficult absorption of coating materials, reduce the anti-ultraviolet ability of nanoparticles, and achieve high total antioxidant capacity , low cost and simple preparation process

Pending Publication Date: 2022-01-21
SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For TiO 2 In the application of nanoparticles in sunscreen, there are reactive oxygen species (ROS) produced by cells and coating materials are not easily absorbed to reduce TiO 2 The problem of anti-ultraviolet ability of nanoparticles, and the oxidation resistance of Zn-HcMT and its combination with TiO have not been reported in literature and patents. 2 Application of nanoparticle complex in sunscreen

Method used

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  • Halostachys caspica metallothionein HcMT compound and application thereof in sunscreen cream
  • Halostachys caspica metallothionein HcMT compound and application thereof in sunscreen cream
  • Halostachys caspica metallothionein HcMT compound and application thereof in sunscreen cream

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Expression and Purification of Zn-Salacia metallothionein HcMT fusion protein

[0032] Transform the expression vector pET-32a-HcMT into competent cells BL21, screen the positive single clones, inoculate the positive single clones in 10 mL of LB medium (Amp final concentration 50 mM) and culture at 37°C, 220 rpm shaker for 8~ 12h, then inoculated into 500mL culture medium according to 1:100(v / v), cultured on a shaker at 37°C and 220rpm for 4h, until the bacterial concentration was OD 600 When reaching 0.6-0.8, add inducer IPTG (final concentration is 0.6mM) and ZnSO 4 (The final concentration is 200 μM) continue to culture for 6h. Centrifuge at 12000rpm for 1min to collect the induced cells, add protein loading buffer, bath in boiling water for 15min, ice bath for 2min, detect protein expression by 15% SDS-PAGE, see attached for specific results figure 1 shown.

[0033] Depend on figure 1 It can be seen that the expressed fusion protein was identified by ...

Embodiment 2

[0035] Embodiment 2: Antioxidative activity test of recombinant protein Zn-HcMT

[0036] (1) Recombinant protein Zn-HcMT scavenging hydroxyl radical test

[0037] Take 8 10mL test tubes, add 9mMFeSO 4 1mL, 9mM salicylic acid-ethanol 1mL, add 1mL of Zn-HcMT solution of different concentrations (0.3114, 0.2803, 0.2491, 0.2180, 0.1868, 0.1557, 0.1246, 0.0934mg / mL), mix well and add the concentration of 8.8mM h 2 o 2 Start the reaction with 1 mL, heat the reaction in a water bath at 37°C for 0.5 h, and measure the absorbance at 510 nm with distilled water as a reference. Set the experimental group (Ax), background light absorption group (Ax0) and blank control group (A0), hydroxyl radical scavenging rate=[A0-(Ax-Ax0)] / A0×100%, each concentration carries out three groups of parallel For the test results, please refer to the appendix image 3 .

[0038] Depend on image 3 The data shows that there is an obvious concentration-dependent relationship between the hydroxyl radic...

Embodiment 3

[0059] Embodiment 3: Zn-Salacia metallothionein HcMT complex

[0060] A Zn-salia metallothionein HcMT complex applied in sunscreen, the complex includes Zn-salia metallothionein HcMT and TiO nanoparticles, Zn-salia metallothionein HcMT complex specific Prepared by following steps:

[0061] (1) Identify the expressed fusion protein by SDS-PAGE, transform the expression vector pET-32a-HcMT into competent cells BL21, screen positive single clones, inoculate positive single clones in 10 mL of LB medium, and incubate at 37°C , cultured in a shaker at 220rpm for 8-12h, then inserted into 500mL culture medium at a volume ratio of 1:100, cultured on a shaker at 220rpm at 37°C for 4h, until the OD600 of the bacterial solution reached 0.6-0.8, and the final concentration was 0.6mM The inducer IPTG and ZnSO at a final concentration of 200 μM 4 Solution, continue to cultivate for 6h, centrifuge at 12000rpm for 1min to collect the induced cells, add protein loading buffer, bath in boilin...

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Abstract

The invention discloses a Zn-halostachys caspica metallothionein (HcMT) compound applied to sunscreen cream, which comprises Zn-halostachys caspica metallothionein (HcMT) and TiO2 nanoparticles. The TiO2-Zn-HcMT compound powder is obtained through SDS-PAGE identification and expression of fusion protein and optimization of a purification process and a preparation process, the oxidation resistance of Zn-HcMT crude protein is investigated, the Zn-HcMT crude protein has high scavenging activity on free radicals such as (.OH), (O2 <->.) and DPPH, the total oxidation resistance is excellent, meanwhile, the Zn-HcMT crude protein has certain reducing power, and by means of the excellent oxidation resistance of the Zn-HcMT crude protein, SEM and FTIR are adopted for characterizing Zn-HcMT / TiO2, the anti-ultraviolet capacity of the Zn-HcMT / TiO2 compound in the sunscreen cream is remarkably improved than that of TiO2 coated with an organic material, the capacity of removing hydroxyl radicals, superoxide anions and DPPH free radicals and the total antioxidant capacity are remarkably improved, it can be seen that compounding of the Zn-HcMT and the TiO2 nanoparticles has the synergistic interaction effect, the problem of application of TiO2 nanoparticles in the sunscreen cream is solved, the application potential in the field of cosmetics is huge, and the compound is an efficient and safe cosmetic active ingredient.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, in particular, the invention relates to the technical field of a salt ear wood metallothionein complex and its application. [0002] technical background [0003] Metallothionein (MT) is a class of cysteine-rich low molecular weight (6-7Kd) multifunctional inducible proteins that are ubiquitous in organisms. Metallothionein is highly conserved in evolution. According to the metal ion content and amino acid composition, it can be divided into four subtypes: MT-I, MT-II, MT-III and MT-IV. MT participates in various important physiological activities in the body, such as detoxification of heavy metals, scavenging free radicals, regulating The concentration of trace elements in the body, and plays an important role in anti-tumor, anti-apoptosis, hormone regulation, cell metabolism, proliferation and differentiation. Since American scholar Mackay first discovered and isolated MT from horse kidne...

Claims

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Application Information

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IPC IPC(8): C12N15/70C12N15/62C12P21/02C07K19/00C07K1/14C07K1/22A61K8/64A61K8/29A61Q17/04
CPCC12N15/70C07K14/825A61K8/645A61K8/29A61Q17/04C07K2319/00
Inventor 刘忠渊刘阳彭翠毛新芳吴彦辰邓夕莞张凤邹培瑜王洁萍
Owner SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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