Yangtze alligator antimicrobial peptide Alligatorin 6 and application thereof

An antibacterial peptide, Chinese alligator technology, applied in the field of biomedicine, can solve the problems of cell death, dissolution of cell contents, destruction of cell membrane integrity, etc.

Active Publication Date: 2014-12-03
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

The bactericidal mechanism of most antimicrobial peptides is by acting on the phospholipid bilayer on the bacterial cell membrane, destroying the int

Method used

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  • Yangtze alligator antimicrobial peptide Alligatorin 6 and application thereof
  • Yangtze alligator antimicrobial peptide Alligatorin 6 and application thereof
  • Yangtze alligator antimicrobial peptide Alligatorin 6 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Discovery of alligator alligator antimicrobial peptide Alligatorin6

[0013] Through retrieval, a gene sequence (XP_006026474.1) with a length of 459bp 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 152 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 (...

Embodiment 2

[0018] Chemical Synthesis of Antimicrobial Peptide Alligatorin6 from Chinese Alligator

[0019] (1) Synthesize the full sequence of Alligatorin6 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 purified Alligatorin6 was identified by high performance liquid chromatography (HPLC) with a purity >95%, its molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF, 3044.5Da), and its isoelectric point was 12.48 by isoelectric focusing electrophoresis , using an automatic amino acid sequencer to determine its amino acid sequence structure is consistent with the natural Chinese alligator antimicrobial peptide Alligatorin6.

Embodiment 3

[0021] Detection of Antibacterial Activity of Chinese Alligator Antimicrobial Peptide Alligatorin6

[0022] (1) Pick the test strains preserved on the slant (Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853 were purchased from the China Common Microorganism Culture Collection Management Center, and other strains were collected from the local hospital) and spread evenly on the MH solid medium (purchased from Qingdao Haibo Biotechnology Co., Ltd.) plate, place a sterilized filter paper sheet with a diameter of 0.5 cm on the surface of the medium, add dropwise 10 μl of Alligatorin6 sample solution of 2 mg / ml 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. ...

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Abstract

The invention discloses a Yangtze alligator antimicrobial peptide Alligatorin 6 as well as an amino acid sequence thereof and an application of the Yangtze alligator antimicrobial peptide Alligatorin 6 in antibiosis. The antimicrobial peptide Alligatorin 6 is a mature peptide sequence of the antimicrobial peptide which is obtained by searching a Yangtze alligator gene database by virtue of screening and analyzing, wherein an amino acid sequence of the mature peptide sequence is as shown in a sequence table SEQ ID NO.1. By virtue of analysis and comparison, the Yangtze alligator antimicrobial peptide Alligatorin 6 has obvious difference from the amino acid sequences of all of the existing known antimicrobial peptides, and belongs to a novel antimicrobial peptide. According to an antimicrobial test, the Yangtze alligator antimicrobial peptide Alligatorin 6 has very strong antimicrobial activity on gram positive bacteria and gram negative bacteria, is broad-spectrum and efficient in antimicrobial action. Besides, the Yangtze alligator antimicrobial peptide Alligatorin 6 has the characteristics of low hemolytic activity, good salt tolerance, good heat tolerance and good heat stability, and can be applied to the fields of medicines, cosmetics, food fresh keeping and breeding industry.

Description

technical field [0001] The invention relates to a natural antibacterial peptide and its application, more specifically, to an antibacterial peptide Alligatorin6 derived from Chinese alligator and its application, belonging 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 i...

Claims

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Application Information

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IPC IPC(8): C07K14/46C07K1/20A61K38/17A61K8/64A61P31/04A23K1/165A23L3/3526
CPCA23L3/3544A61K8/64A61K38/00A23K20/147C07K14/46
Inventor 王义鹏于海宁蔡莎莎高久香
Owner 宜肌坊(厦门)生物科技有限公司
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