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Natural antibacterial peptide and application thereof

A natural antibacterial and antimicrobial peptide technology, applied in the field of biomedicine, can solve problems such as antibiotic resistance of pathogenic microorganisms, threats to human health, etc., and achieve the effects of good stability, small molecular weight, and wide application prospects

Active Publication Date: 2015-04-08
王义鹏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] In recent years, the large-scale abuse of traditional antibiotics has led to more and more serious drug resistance of pathogenic microorganisms, posing a huge threat to human health.

Method used

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  • Natural antibacterial peptide and application thereof
  • Natural antibacterial peptide and application thereof
  • Natural antibacterial peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Cloning of natural antimicrobial peptide QHA coding gene:

[0016] 1) Extraction of total RNA from the venom glands of Qinghuan sea snakes:

[0017] ①Take 250 mg of venomous tissue of sea snake qinghuan, put it into a mortar and add liquid nitrogen to grind it into powder, transfer it to an EP tube, add 1 ml of total RNA extraction reagent (Trizol, product of Invitrogen, USA), mix well, and then incubate at 4°C , centrifuged at 12000rpm for 10min.

[0018] ② Centrifuge to take the supernatant, add 0.2ml chloroform solution, mix vigorously, let stand at room temperature for 10 minutes, then centrifuge at 4°C, 12000rpm for 10 minutes, and discard the precipitate.

[0019] ③ Add an equal volume of isopropanol to the supernatant, place it at room temperature for 10 minutes, centrifuge at 4°C, 12000rpm for 10 minutes, collect the precipitate and wash it once with 75% (V / V) ethanol, dry it, and the precipitate at the bottom of the tube is the blue ring Sea snake venom gland...

Embodiment 2

[0035] Chemical synthesis of antimicrobial peptide QHA:

[0036]Ⅰ. The chemical synthesis method of the antimicrobial peptide QHA: According to the amino acid sequence of the mature peptide deduced from the gene, its complete sequence was synthesized with an automatic peptide synthesizer (433A, Applied Biosystems), and desalted by HPLC reverse-phase column chromatography.

[0037] Ⅱ. Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0038] Ⅲ. The purity of the purified antibacterial peptide QHA was identified by high performance liquid chromatography (HPLC), the molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), the isoelectric point was determined by isoelectric focusing electrophoresis, and an automatic amino acid sequencer was used to determine Amino acid sequence structure.

[0039] The antimicrobial peptide QHA is a linear...

Embodiment 3

[0042] Detection of antibacterial peptide QHA antibacterial activity:

[0043] (1) Pick the test strains stored on the slant and evenly spread them on the MH solid medium (purchased from Qingdao Haibo Biotechnology Co., Ltd.) plate, and place the sterilized 0.5cm diameter filter paper on the medium On the surface, drop 10 μl of 2 mg / ml antimicrobial peptide QHA sample solution dissolved in sterilized deionized water, incubate upside down at 37°C for 18-20 hours, and observe whether the inhibition zone is formed or not. 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.

[0044] (2) Antimicrobial peptide QHA minimum inhibitory concentration (Minimum Inhibitory Concentration) determination (2-fold dilution method):

[0045] Select the strains with the inhibition zone in the previous experiment to carry out the ...

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Abstract

Belonging to the technical field of biomedicine, the invention in particular relates to a natural antibacterial peptide QHA and application thereof. The natural antibacterial peptide QHA is composed of 30 amino acid residues, is straight chain polypeptide, and has a molecular weight of 3628.5Da and isoelectric point of 12.61. The antibacterial peptide QHA provided by the invention has the beneficial characteristics of small molecular weight, simple chemical synthesis method, efficient, broad-spectrum and efficient antibacterial action, and good stability, etc., thus having broad application prospects. In addition, the invention also relates to a coding gene of antibacterial peptide QHA and application thereof.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a natural antibacterial peptide QHA and its application. 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. Antimicrobial peptides have the advantages of small molecular weight, simple structure, strong antibacterial activity...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46C12N15/12A61K38/17A61K8/64A61P31/10A61P31/04A23K1/17A01N47/44A01P1/00A01P3/00A23K20/195
CPCA61K8/64A61K38/00C07K14/46
Inventor 王义鹏
Owner 王义鹏
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