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A kind of antimicrobial peptide based on cell-penetrating peptide tat(49-57) and its synthesis method

A synthesis method and technology of antimicrobial peptides, applied in the field of antimicrobial peptides based on cell-penetrating peptide Tat and its synthesis, can solve problems such as unreasonable use, multi-drug resistant bacteria, death, etc., and achieve good inhibition efficiency

Inactive Publication Date: 2020-09-04
ZHENGZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the irrational use of antibiotics leads to the subsequent emergence of multi-drug resistant bacteria and the death caused by drug-resistant bacterial infection has become a worldwide public health problem. For this reason, new antibiotics that can resist multi-drug resistant bacteria are urgently needed

Method used

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  • A kind of antimicrobial peptide based on cell-penetrating peptide tat(49-57) and its synthesis method
  • A kind of antimicrobial peptide based on cell-penetrating peptide tat(49-57) and its synthesis method
  • A kind of antimicrobial peptide based on cell-penetrating peptide tat(49-57) and its synthesis method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1--the synthesis of antimicrobial peptide

[0026] 1 Experimental part

[0027] 1.1. Experimental reagents

[0028]

[0029] Preparation of ninhydrin detection solution: Weigh 1 g of ninhydrin and fully dissolve it in 20 mL of absolute ethanol, and the yellow solution formed is the ninhydrin detection solution.

[0030] Preparation of deprotection reagent: Mix 20 mL of piperidine in 80 mL of DMF to make a 20% piperidine DMF solution as the deprotection reagent.

[0031] Preparation of cutting reagent: Mix trifluoroacetic acid, thioanisole, ethanedithiol, water and phenol in a volume ratio of 82.5:5:5:5:2.5, and the mixture is the cutting reagent.

[0032] 1.2 Experimental Instruments

[0033]

[0034] 2. Experimental steps and methods

[0035] 2.1 Solid-phase synthesis of antimicrobial peptides

[0036] The peptide chain sequences of Tat(49-57) and its derived peptides Tat(YG), Tat(YY), Tat(FG), and Tat(FF) are RKKRRQRRR (as shown in SEQ ID No.1), ...

Embodiment 2

[0068] Embodiment 2--antibacterial activity detection of polypeptide

[0069] 1. Experimental Materials and Instruments

[0070] 1.1 Bacteria used in the experiment

[0071] Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Salmonella typhimurium were all commercially available.

[0072] 1.2 Main reagents

[0073]

[0074] 1.3 Main Instruments

[0075]

[0076] 2 Experimental methods

[0077] 2.1 Preparation of main solution

[0078] LB liquid medium: Take 10 g of peptone, 10 g of sodium chloride, and 5 g of yeast powder in a large beaker, add 1000 mL of distilled water, and adjust the pH to 7.2-7.4 with concentrated sodium hydroxide solution after dissolution. The prepared medium was sterilized by moist heat at 121 °C for 20 min, then distributed into 250 mL Erlenmeyer flasks, and stored at room temperature until use.

[0079] MTT test solution: Weigh 100 mg MTT and dissolve it in 20 mL sodium phosphate buffer (pH=7.0), filter through a sterile filte...

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Abstract

The invention belongs to the field of antimicrobial peptides, and discloses an antimicrobial peptide based on a cell penetrating peptide Tat (49-57) and a synthetic method of the antimicrobial peptide. The antimicrobial peptide is Tat (YG), Tat (YY), Tat (FG) or Tat (FF), and a sequence of a peptide chain is successively as shown by SEQ ID No. 2 to No. 5. The antimicrobial peptide is synthesized by adopting Wang resin as a carrier, adopting Fmoc as an amino acid side chain protection base, adopting a DMF solution of piperidine as a deprotection reagent and adopting HBTU, HOBT and DIEA as amino acid condensing agents in a solid-phase synthetic method. The four derivative antimicrobial peptides have good inhibition efficiency for escherichia coli, salmonella typhimurium, Bacillus subtilis and staphylococcus aureus, and the inhibition rate for some bacteria can be higher than that of the cell penetrating peptide Tat (49-57); and moreover, the hemolytic activity of the four derivative microbial peptides is small, and the concentration when the bacteriostatic activity is played cannot reach the lowest hemolytic concentration.

Description

technical field [0001] The invention belongs to the field of antibacterial peptides, and in particular relates to an antibacterial peptide based on cell penetrating peptide Tat (49-57) and a synthesis method thereof. Background technique [0002] Antibiotics play an important role in the prevention and treatment of diseases caused by microbial infections. However, the irrational use of antibiotics leading to the subsequent emergence of multi-drug resistant bacteria and the death caused by drug-resistant bacterial infections has become a worldwide public health problem. For this reason, new antibiotics that can resist multi-drug resistant bacteria are urgently needed. The unique antibacterial mechanism of antimicrobial peptides, the lack of drug resistance, and the non-toxicity to normal cells make them promising to become a new generation of effective antibacterial drugs, and their application prospects and value have attracted widespread attention. Studying the antibacteri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/16C07K1/06C07K1/04C07K1/20A61P31/04
CPCC07K14/005C12N2740/16022Y02A50/30Y02P20/55
Inventor 吕名秀李林璐卢奎孙志杰赵玉芬段冰潮
Owner ZHENGZHOU UNIV
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