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Preparation and application of antiglycation oligopeptide-5 (AdP)

An anti-glycation and oligopeptide technology is applied in the preparation methods of peptides, cosmetic preparations, preparations for skin care, etc., to achieve the effects of inhibiting protein glycosylation, low equipment cost and simple operation.

Pending Publication Date: 2022-01-18
守正创新生物科技(天津)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to apply genetic engineering technology to provide a preparation and application of anti-glycosylated oligopeptide-5 (AdP), so as to solve the technical problem of producing oligopeptide-5 (AdP) by traditional means

Method used

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  • Preparation and application of antiglycation oligopeptide-5 (AdP)
  • Preparation and application of antiglycation oligopeptide-5 (AdP)
  • Preparation and application of antiglycation oligopeptide-5 (AdP)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The activated recombinant expression strain ER2566-pET28a-AdP was inoculated in the LB medium containing Kan resistance according to the addition amount of 3%, 27°C, 100rpm, shaker culture, when A600=1.0. Put the seed medium into the fermenter according to the inoculation amount of 10%, set the temperature at 27°C, rotate at 200rpm, adjust the pH with ammonia water, when A600=7.0, add 5mL of 0.5mmol / L IPTG for induction, and induce for 5h Afterwards, the target protein AdP is expressed in the strain.

[0034]The fermentation broth was centrifuged at 8,000 rpm for 30 min at 4°C with a refrigerated centrifuge, the supernatant was discarded, and the precipitate was stored at 4°C for future use. Take an appropriate amount of precipitate, resuspend the dispersed bacteria with 20 times the amount of Tris-Hcl buffer, process it once with a high-pressure homogenizer, and then centrifuge at 8000rpm for 30min at 2°C in a refrigerated centrifuge to collect the supernatant. Use a ...

Embodiment 2

[0036] The activated recombinant expression strain ER2566-pET28a-AdP was inoculated in the LB medium containing Kan resistance according to the addition amount of 1%, 37°C, 200rpm, shaker culture, when A600=1.0~1.5. Put the seed medium into the fermenter according to the inoculation amount of 8%, set the temperature at 37°C, rotate at 300rpm, adjust the pH with ammonia water, when A600=7.8, add 7mL of 0.5mmol / L IPTG for induction, and induce for 3h Afterwards, the target protein AdP is expressed in the strain.

[0037] The fermentation broth was centrifuged at 12,000 rpm for 25 min at 4°C with a refrigerated centrifuge, the supernatant was discarded, and the precipitate was stored at 4°C for future use. Take an appropriate amount of precipitate, resuspend and disperse the bacteria with 20 times the amount of Tris-Hcl buffer, process 3 times with a high-pressure homogenizer, and then use a refrigerated centrifuge at 12,000 rpm for 25 minutes at 4°C to collect the supernatant. ...

Embodiment 3

[0039] The activated recombinant expression strain ER2566-pET28a-AdP was inoculated in the LB medium containing Kan resistance according to the addition amount of 1%, cultured on a shaker at 42°C, 100 rpm, when A600=2.0. Put the seed medium into the fermenter according to the inoculation amount of 6%, set the temperature at 42°C, rotate at 500rpm, adjust the pH with ammonia water, when A600=9.0, add 10mL of 0.5mmol / L IPTG for induction, and induce for 2h Afterwards, the target protein AdP is expressed in the strain.

[0040] The fermentation broth was centrifuged at 15,000 rpm for 10 min at 4°C with a refrigerated centrifuge, the supernatant was discarded, and the precipitate was stored at 4°C for future use. Take an appropriate amount of precipitate, resuspend and disperse the bacteria with 20 times the amount of Tris-Hcl buffer, process 5 times with a high-pressure homogenizer, and then use a refrigerated centrifuge at 10°C at a speed of 15,000 rpm for 10 minutes to collect ...

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Abstract

The invention relates to preparation and application of antiglycation oligopeptide-5 (AdP), AdP is produced by adopting a genetic engineering means, the preparation comprises the following steps: carrier construction and screening, seed solution preparation, induced expression, separation and purification, etc., compared with a traditional biological peptide synthesis method, the preparation has the characteristics of small pollution, low equipment cost and simple operation, and the produced AdP is closer to natural AdP in structure, better in activity and higher in purity; and AdP is mainly prepared from alanine, arginine, aspartic acid, glutamic acid, glycine, histidine, etc., AdP can promote generation of fibroblasts and keep skin vitality, has a good anti-aging effect, and can inhibit melanogenesis, it is found for the first time that AdP can protect macromolecular protein and effectively prevent the occurrence of protein glycosylation, and AdP has multiple effects of moisturizing, repairing, skin pigment removing and brightening, wrinkle removing and glycation resisting.

Description

technical field [0001] The invention belongs to the field of cosmetics, and in particular relates to an oligopeptide-5 (AdP) with anti-glycation effect and a preparation method thereof. Background technique [0002] Oligopeptide-5 (AdP) is an oligopeptide derived from aminoacyl-tRNA synthetase-interacting multi-functional protein 1 (AIMP1). [0003] AIMP1, also known as p43, MSC p43 or proEMAPII, is a fairly conserved protein in mammals, expressed in almost all cells, and was first discovered in a huge aminoacyl-tRNA synthetase (aaRS ) center of the complex, as a non-enzymatic auxiliary protein, it participates in the formation of the complex and plays a key role in maintaining the structure of the complex. AIMP1 is widely distributed in the nucleus, cytoplasm, and endoplasmic reticulum. Different subcellular locations have different functions, such as participating in protein synthesis, regulating autoimmune responses, etc., and can also be secreted as a cytokine to partic...

Claims

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Application Information

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IPC IPC(8): C12P21/02C07K14/435C07K1/34C07K1/18C07K1/36A61K8/64A61Q7/00A61Q17/04A61Q19/00A61Q19/02A61Q19/08C12R1/19
CPCC12P21/02C07K14/435A61K8/64A61Q19/00A61Q19/02A61Q19/08A61Q19/005A61Q17/04A61Q19/008A61Q7/00
Inventor 杨秀芬王培培郑春阳
Owner 守正创新生物科技(天津)有限公司
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