Modified antibacterial peptide C-CM8 of Chelonia mydas antibacterial peptide and preparation method and application thereof

A C-CM8, antibacterial peptide technology, applied in the field of biomedicine, can solve problems such as drug resistance of pathogenic microorganisms, and achieve the effect of strong anti-inflammatory activity, small molecular weight, broad-spectrum and high-efficiency antibacterial activity

Pending Publication Date: 2021-04-09
宜肌坊(厦门)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the abuse of traditional antibiotics leads to more and more serious drug resistance of pathogenic microorganisms, and to provide a modified antimicrobial peptide C-CM8 of the green sea turtle (Chelonia mydas) antimicrobial peptide Cm-CATH2 and its preparation method and application

Method used

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  • Modified antibacterial peptide C-CM8 of Chelonia mydas antibacterial peptide and preparation method and application thereof
  • Modified antibacterial peptide C-CM8 of Chelonia mydas antibacterial peptide and preparation method and application thereof
  • Modified antibacterial peptide C-CM8 of Chelonia mydas antibacterial peptide and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The chemical synthesis of the modified antimicrobial peptide C-CM8 of the green sea turtle antimicrobial peptide Cm-CATH2 is as follows.

[0022] According to the amino acid sequence and secondary structure of the green sea turtle antimicrobial peptide Cm-CATH2, a fragment of 17 amino acids at the N-terminus was intercepted by using the molecular transformation method of sequence cutting, and the α-helical region of the N-terminus was partially retained, and its C-terminus was completely removed. The modified antimicrobial peptide C-CM5 was designed and obtained without a regular structure, and it was chemically synthesized by the method of peptide solid-phase synthesis. The specific preparation method is as follows:

[0023] 1. According to the amino acid sequence of the modified antimicrobial peptide C-CM8 obtained by design, its complete sequence was synthesized with an automatic peptide synthesizer (433A, Applied Biosystems), and purified by desalting and purifying b...

Embodiment 2

[0030] Determination of antibacterial activity of modified antimicrobial peptide C-CM8:

[0031] Pick the test strains stored on the slant respectively, spread them evenly on the plate of MH solid culture medium (Beijing Suo Laibao Technology Co., Ltd.), place the sterilized 0.5cm diameter filter paper on the surface of the culture medium, add dropwise Dissolve 10 μl of 2 mg / ml antimicrobial peptide C-CM8 sample solution in sterilized deionized water, incubate upside down at 37°C for 18-20 hours, and observe whether the inhibition zone is formed or not.

[0032] If the sample has antibacterial activity, a clear and transparent bacteriostatic zone will be formed around the filter paper, and the larger the bacteriostatic zone, the stronger the antibacterial activity of the sample.

[0033] (2) Determination of Antimicrobial Peptide C-CM8 Minimum Inhibitory Concentration (2-fold dilution method):

[0034] Select the strains with the inhibition zone in the previous experiment to ...

Embodiment 3

[0044] Determination of hemolytic activity of modified antimicrobial peptide C-CM8:

[0045] The collected rabbit blood was mixed with Alfred's solution for anticoagulation, washed twice with normal saline and resuspended into a suspension of 107-108cell / ml.

[0046] The above-mentioned diluted erythrocyte suspension was mixed with the modified antimicrobial peptide C-CM8 sample dissolved in normal saline, incubated at 37°C for 30 minutes, then centrifuged at 1000rpm for 5 minutes, and the supernatant was measured for absorbance at 540nm. Physiological saline was used as the negative control, 10% Triton X-100 was used as the positive control, and the hemolysis percentage was calculated according to the following formula: hemolysis percentage H%=A sample-A negative control / A positive control×100%.

[0047]The results showed that the sample concentration was 100 μg / ml, and the hemolysis percentage of the modified antimicrobial peptide C-CM8 was 1.34%. It shows that the modified...

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Abstract

The invention provides a modified antibacterial peptide C-CM8 of a Chelonia mydas antibacterial peptide and a preparation method and application thereof. The N end of the modified antibacterial peptide is an alpha helix, the C end is a polypeptide of an irregular structure, the modified antibacterial peptide contains 17 amino acid residues, the molecular weight is 1998.37Da, and the isoelectric point is 12.60. The modified antibacterial peptide C-CM8 has broad-spectrum and efficient antibacterial activity and extremely high anti-inflammatory activity, has the advantages of being small in molecular weight, simple in structure, low in hemolytic activity, simple in preparation method and the like, is expected to become a novel efficient antibacterial drug, has wide development and application prospects, and is especially used for preparing antibacterial drugs or compositions, bacteria growth inhibiting drugs or compositions, anti-inflammatory drugs or compositions, preservatives, animal feed additives or cosmetic additives.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a modified antimicrobial peptide C-CM8 of a green sea turtle (Chelonia mydas) antimicrobial peptide Cm-CATH2 and a preparation method and application thereof. Background technique [0002] In recent years, the large-scale abuse of traditional antibiotics has led to more and more serious problems of drug resistance of pathogenic microorganisms, posing a huge threat to human health. The clinical response to drug-resistant microbial infections is to use new or alternative antibiotics that have not been used against drug-resistant microorganisms, so this requires the continuous development of new antimicrobial drugs. [0003] Antimicrobial peptides are natural small molecular polypeptides encoded by the genes of organisms, and are important molecules in the immune system of organisms. They have direct killing effects on bacteria, fungi, viruses and even protozoa. Ant...

Claims

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Application Information

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IPC IPC(8): C07K14/46C07K1/04C07K1/06C07K1/20A61K38/17A61P31/04A61P31/10A61P29/00A61K8/64A61Q19/00A23K20/147
CPCC07K14/46A61P31/04A61P31/10A61P29/00A61K8/64A61Q19/00A23K20/147A61K38/00A61K2800/10
Inventor 孙碧昌王义鹏于海宁欧阳建红
Owner 宜肌坊(厦门)生物科技有限公司
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