Proline-rich antibacterial peptide Pyr-2 targeting gram-negative bacteria as well as preparation method and application thereof

A Gram-negative bacteria, proline-rich technology, applied in the preparation method of peptides, chemical instruments and methods, medical preparations containing active ingredients, etc., can solve the complex synthesis process, high synthesis cost, peptide hemolytic activity Advanced problems, to achieve the effect of high application value, good biocompatibility, and simple technical route

Active Publication Date: 2021-10-26
NORTHEAST AGRICULTURAL UNIVERSITY
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The natural antimicrobial peptide Pyrrhocoricin is a proline-rich antimicrobial peptide that exists in insects. It targets bacterial ribosomes and only has antibacterial activity against Gram-negative bacteria. How

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Proline-rich antibacterial peptide Pyr-2 targeting gram-negative bacteria as well as preparation method and application thereof
  • Proline-rich antibacterial peptide Pyr-2 targeting gram-negative bacteria as well as preparation method and application thereof
  • Proline-rich antibacterial peptide Pyr-2 targeting gram-negative bacteria as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0017] Example 1

[0018] Natural antibacterial peptide pyrhocoricin from insects is replaced by amino acid, and the proline is replaced with the fine-hydrophilic amino acid in the sequence, and the number of proline is increased to increase the antibacterial peptide hydrophilicity; and use fine The amine is replaced with threonine in the 11th position and increases the positive charge of the antibacterial peptide. The sequence and physical and chemical parameters of PYR-2 are shown in Table 1.

[0019] Table 1 peptide amino acid sequence and physical and chemical parameters

[0020]

Example Embodiment

[0021] Example 2

[0022] Synthesis of antimicrobial peptides in solid phase chemical synthesis

[0023] 1. Preparation of antimicrobial peptides is carried out from the C-terminal to N port, and is accomplished by a polypeptide synthesizer. First, FMOC-X (X is the first amino acid of each antimicrobial peptide) to the WANG resin, then remove the Fmoc group to obtain x-wang resin; FMOC-Y-TRT-OH (9 - The methyl, y, y is the second amino acid of each antimicrobial peptide C-terminal; according to this procedure, it is synthesized from the C-terminal to the N-terminal until the synthesis is completed, and the side of the FMOC group is obtained. Chain protection resin;

[0024] 2, in the above-mentioned peptide resin, a cutting reagent is added to 20 ° C. 2H, filter; filtration; precipitated TFA (trifluoroacetic acid) washing, mixing the lotion with the above filtrate, rotary evaporator concentrate, then add 10 times The left and right volume of predherent water-free ether, -20 ° C pr...

Example Embodiment

[0027] Example 3

[0028] Determination of antibacterial peptide antibacterial activity

[0029] 1, minimum antibacterial concentration assay: the minimum antibacterial concentration (MIC) of the peptide was determined using a standard micro-dilution dilution method. The bacteria incubated at 220 rpm to incubate overnight at 37 ° C, then transferred to new MHB until the growth of the alignment period, and dilute the bacteria to 10 5 CFU / ml. 50 μl of different concentrations of peptide (peptide had a final concentration of 1-128 μm) and 50 μl of bacterial suspension were added to each well of 96-well plates. The negative control is an immigaptic MHB medium, a positive control as a MHB medium of gegrotia, and the 96-well plate is placed in a 37 ° C constant temperature incubator for 18 to 20 hours. The antibacterial peptide concentration of microbial growth was observed under the naked eye and spectrophotometry. Three independent repetition tests, each repeating two parallel. The ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a proline-rich antibacterial peptide Pyr-2 targeting gram-negative bacteria as well as a preparation method and application thereof. The sequence of the antibacterial peptide is as shown in SEQ ID No. 1. By taking amino acids at the 2nd to 10th sites in a natural antibacterial peptide pyrrhocoricin as a core sequence, glycine and serine at the 4th and 5th sites in the sequence are respectively replaced by utilizing proline, so as to increase the quantity of proline and improve the hydrophobicity; and threonine at the 11th site is replaced by arginine, so that the positive charge quantity of the polypeptide is increased, and the antibacterial activity of the polypeptide is improved. The antibacterial peptide Pyr-2 is applied in preparation of drugs for treating infectious diseases caused by gram-negative bacteria. The antibacterial peptide Pyr-2 provided by the invention almost has no hemolysis on red blood cells, and has good biocompatibility. In conclusion, the Pyr-2 disclosed by the invention has the potential of becoming an antibacterial drug for specifically resisting gram-negative bacterium infection, and has a very high application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a proline-rich antimicrobial peptide Pyr-2 targeting Gram-negative bacteria, a preparation method and application thereof. Background technique [0002] In the past few decades, the abuse of antibiotics has led to the continuous strengthening of bacterial resistance, and the number of resistant bacteria has also increased. In order to prevent the spread and infection of these bacteria, it has become a priority to develop new drugs that can replace antibiotics. It is a problem that people urgently need to solve in recent years. Antimicrobial peptides (AMPs) are a class of polypeptide substances with broad-spectrum antibacterial properties, and are important components of the innate immune system; they exist in a wide range and are found in animals and plants. Compared with antibiotics, antimicrobial peptides have the advantages of non-toxic side effects (only a small part...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C07K14/435C07K1/04C07K1/20C07K1/14A61K38/10A61P31/04
CPCC07K14/43563A61P31/04A61K38/00Y02A50/30
Inventor 吕银凤谭美姝张丽聪马清泉陈婷婷单安山杨城义
Owner NORTHEAST AGRICULTURAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap