Reptile antibacterial peptide Alligatorin5 and applications thereof

A technology of reptiles and antimicrobial peptides, applied in the field of biomedicine, can solve problems such as cell death, damage to cell membrane integrity, and dissolution of cell contents

Active Publication Date: 2014-11-26
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The bactericidal mechanism of most antimicrobial peptides is by acting on the phospholipid bilayer on the bacterial cell membrane, destroying the integrity of the cell membrane and forming a transmembrane channel on the cell membrane, resulting in the dissolution of the cell content and cell death

Method used

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  • Reptile antibacterial peptide Alligatorin5 and applications thereof
  • Reptile antibacterial peptide Alligatorin5 and applications thereof
  • Reptile antibacterial peptide Alligatorin5 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Discovery of reptile antimicrobial peptide Alligatorin5

[0013] Through retrieval, a gene sequence (XP_006026473.1) with a length of 477bp was found from the Chinese alligator genome data on the website of the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov) , the encoded polypeptide precursor consists of 158 amino acid residues. The gene and its encoded polypeptide precursor are respectively shown in SEQ ID NO: 2 and SEQ ID NO: 3:

[0014]

[0015]

[0016] The blastX software (http: / / blast.st-va.ncbi.nlm.nih.gov / Blast.cgi?PROGRAM=blastx&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome) of the gene sequence on the NCBI website was used for sequence comparison and analysis, and the results showed that the The encoded product of the gene may be the precursor of antimicrobial peptides of the alligator cathelicidins family. The polypeptide precursor sequence encoded by the gene was further sequenced using the protein blast software (http: / / blast...

Embodiment 2

[0018] Chemical Synthesis of Antimicrobial Peptide Alligatorin5

[0019] (1) Synthesize the full sequence of Alligatorin5 with an automatic peptide synthesizer (433A, Applied Biosystems), and pass HPLC C 18 Desalting and purification by reverse phase column chromatography. (2) The molecular weight was determined by conventional matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). (3) The purity of the purified Alligatorin5 was identified by high performance liquid chromatography (HPLC)>95%, the molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF, 2557.1 Da), and the isoelectric point was 12.03 by isoelectric focusing electrophoresis , using an automatic amino acid sequencer to determine its amino acid sequence structure is consistent with the natural Alligatorin5.

Embodiment 3

[0021] Detection of antibacterial activity of antimicrobial peptide Alligatorin5

[0022] (1) Pick the test strains preserved on the slant (Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853 and Staphylococcus aureus ATCC27853 were purchased from China General Microorganism Culture Collection Management Center, and other strains were collected from local hospitals) and spread evenly Spread on the MH solid medium (purchased from Qingdao Haibo Biotechnology Co., Ltd.) plate, place a sterilized filter paper piece with a diameter of 0.5 cm on the surface of the medium, and add 2 mg / ml dissolved in sterilized deionized water dropwise. Alligatorin5 sample solution 10μl, incubate upside down at 37°C for 18-20 hours, 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...

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Abstract

The invention discloses a reptile-derived antibacterial peptide Alligatorin5, an amino acid sequence thereof and applications of the antibacterial peptide in the antibacterial field. The antibacterial peptide Alligatorin5 is derived from Chinese alligator. By searching a Chinese alligator gene database, screening and analyzing, a mature peptide sequence of the antibacterial peptide is obtained. The amino acid sequence of the antibacterial peptide Alligatorin5 is shown as SEQ ID NO.1 in a sequence table. By comparative analysis, the antibacterial peptide Alligatorin5 is obviously different from amino acid sequences of antibacterial peptides known at present, and belongs to a novel antibacterial pepetide. Antibacterial experiments show that: the antibacterial peptide Alligatorin5 has high antibacterial activity for gram positive bacteria, gram negative bacteria and funguses, and is broad in antibacterial spectrum and efficient; the antibacterial peptide Alligatorin5 has characteristics of low hemolytic activity, salt tolerance, heat tolerance, good thermal stability, and the like, and can be used for the fields of medicines, cosmetics, food fresh-keeping and the breeding industry.

Description

technical field [0001] The invention relates to a reptile-derived antimicrobial peptide and its application, more specifically, relates to a reptile-derived alligator antibacterial peptide and its application, and belongs to the technical field of biomedicine. Background technique [0002] Traditional antibiotics have achieved remarkable results in the treatment of bacterial infectious diseases, but in recent years, with the abuse of traditional antibiotics in the fields of medicine and aquaculture, microorganisms have become increasingly resistant to traditional antibiotics, and microbial resistance It has become a difficult problem that seriously threatens human health. The response to microbial resistance has always been to use new or alternative antimicrobial agents that have not been used against drug-resistant microorganisms. Drug resistance, thus requiring the continuous development of new antimicrobial agents. Developing a drug with a new antibacterial mechanism is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46A61K38/17A61K8/64A61P31/04A61P31/10A61Q19/00A23K1/17A23L3/3526A23K20/195
CPCC07K14/46
Inventor 王义鹏于海宁陈燕卫林
Owner SUZHOU UNIV
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