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Antimicrobial peptide separated from skin of Northeast China brown frog and applications in antibacterials

An antimicrobial peptide, a technology for isolation, applied in the field of biomedicine, can solve the problem of less research on skin active peptides

Inactive Publication Date: 2008-12-31
LIAONING UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The application of amphibian drugs has a long history in my country, but the research on their active ingredients and pharmacological properties mainly focuses on small organic molecules such as alkaloids, and there are few researches on their skin active peptides

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of the antimicrobial peptide dybowskin-1CDYa isolated from the skin of Rana sinensis

[0024] Wash the living Rana urana with water, put it in a glass container with a cover, and stimulate it with 8V direct current. It can be seen that a large amount of foamy substances are secreted from the skin of Rana neraeus. The secretion is collected, and acetic acid is added immediately to make the final concentration 4%. , acidified, freeze-dried, after SephadexA-50 chromatography, collected peak II, concentrated, freeze-dried, lyophilized product was subjected to SephadexA-25 chromatography, collected IV peak, concentrated, freeze-dried, and then reversed-phase high performance liquid phase Chromatography (HPLC) C18 column separation and purification, take the peak IV of PT-HPLC, which is the antibacterial peptide dybowskin-1CDYa isolated from the skin of Northeast Rana.

[0025] The purity was identified by PT-HPLC method. The molecular weight was determ...

Embodiment 2

[0026] Example 2 Preparation of antimicrobial peptide dybowskin-2CDYa isolated from Rana skin

[0027] Wash the living Rana urana with water, put it in a glass container with a cover, and stimulate it with 8V direct current. It can be seen that a large amount of foamy substances are secreted from the skin of Rana neraeus. The secretion is collected, and acetic acid is added immediately to make the final concentration 4%. , acidified, freeze-dried, after SephadexG-50 chromatography, collected peak II, concentrated, freeze-dried, lyophilized product was subjected to SephadexA-25 chromatography, collected IV peak, concentrated, freeze-dried, and then reversed-phase high performance liquid phase Chromatography (HPLC) C18 column separation and purification, take the peak VII of PT-HPLC, which is the antimicrobial peptide dybowskin-2CDYa isolated from the skin of Northeast Rana.

[0028] The purity was identified by PT-HPLC method. The molecular weight was determined by fast atom b...

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PUM

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Abstract

The invention relates to an antimicrobial peptide separated from the skin of the northeast Rana chensinensis and the application of the antimicrobial peptide in antibacterial drugs. The technical proposal is that two antimicrobial peptides are sequence-specific single-chain polypeptides separated from the amphibian skin secretions of the northeast Rana chensinensis; the two molecular weight values are 1459.9 and 2184.2 respectively; the two isoelectric points are 9.70 and 12.60, respectively; the amino acids sequences are SEQ ID NO.1 and SEQ ID NO.2 respectively. The preparation method is as follows: the skin secretions of the northeast Rana chensinensis are collected; after the sedimentation is removed through centrifugation, the skin secretions are frozen and dried, and then separated and purified through G50 and G25 molecular sieve chromatography and reversed-phase high pressure liquid chromatography to get a single composition. The invention can be applied to the preparation of the drugs used in the treatment of microbial infections.

Description

Technical field: [0001] The invention belongs to the field of biomedicine, and in particular relates to the isolation of antibacterial peptides from the skin of Rana sinensis and its application in antibacterial drugs. Background technique: [0002] Since the invention of antibiotics, humans have made great achievements in the control and treatment of microbial infection diseases, but with the continuous use of drugs, microbial resistance has become a major problem in the treatment of clinical microbial infection diseases, so that some pathogenic bacteria There is no first-line drug for clinical treatment. Vancomycin-resistant staphylococci, enterococci, and other Gram-negative infections are currently a worldwide clinical problem. The three types of bacteria that mainly cause meningitis also have strong drug resistance clinically. Penicillin-resistant, chloramphenicol-resistant meningococci, pneumococci, and pneumococci resistant to new cephalosporin antibiotics also widel...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/20C07K1/16C07K1/14A61K38/10A61P31/00
Inventor 王秋雨金莉莉宋树森刘剑利
Owner LIAONING UNIVERSITY
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