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Antimicrobial peptide TgHbpe and mutant thereof

An antibacterial peptide and antibacterial drug technology, applied in the field of applied marine biology, can solve the problems of super bacteria, drug resistance of organisms, and great damage to the human body.

Active Publication Date: 2021-03-09
ZHEJIANG WANLI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if antibiotics are used for a long time, the organisms will develop drug resistance, super bacteria will be produced, and the damage to the human body will be greater, which will directly destroy the body's immune system and reduce the body's ability to resist diseases.

Method used

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  • Antimicrobial peptide TgHbpe and mutant thereof
  • Antimicrobial peptide TgHbpe and mutant thereof
  • Antimicrobial peptide TgHbpe and mutant thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Prediction of antimicrobial peptides in clam hemoglobin

[0021] The two subunit sequences of HbI and HbII of cockle hemoglobin were predicted and screened by three online software AntiBP Server, CAMP, and APD2, and six antimicrobial peptides with the highest probability were obtained, and their sequences are shown in Table 1.

[0022] Table 1 Prediction of antimicrobial peptides derived from clam hemoglobin

[0023]

[0024]

Embodiment 2

[0025] Embodiment 2: the selection of antimicrobial peptide

[0026] The antimicrobial peptides used in the experiment (corresponding to the 6 species in Table 1) were synthesized by Hefei Guopeptide Biotechnology Co., Ltd. The N-terminus of the peptide was acetylated and the C-terminus was amidated. The purity was >95%.

[0027] 2.1 Operation steps for selection of antimicrobial peptides

[0028] (1) Take a little bacterial solution of two pathogenic bacteria (Escherichia coli, Vibrio alginolyticus) stored in a refrigerator at -80°C, streak on an LB plate for activation, and select a single colony to inoculate in 15 mL of LB liquid medium , placed on a constant temperature shaker at 37°C for 4-6h until the logarithmic growth phase;

[0029] (2) Take 100 μL of the activated bacterial solution in a 1.5mL centrifuge tube, then add 900 μL of sterilized PBS to dilute, that is, dilute 10 times, and dilute sequentially until it is diluted to 10 6-7 CFU / mL;

[0030] (3) The synthe...

Embodiment 3

[0033] Embodiment 3: Study the secondary and tertiary structure of antimicrobial peptide TgHbpe

[0034] (1) 3D model prediction of antimicrobial peptide TgHbpe

[0035] Use the online software SWISS-MODE to predict the three-dimensional structure of the amino acid sequence of clam hemoglobin HbIIB, select the three-dimensional structure diagram with high similarity, download the pdb file, open it in PyMol software for viewing and editing, and mark the sequence of the antimicrobial peptide TgHbpe , get the 3D model of the antibacterial peptide TgHbpe as shown in figure 1 shown.

[0036] (2) Prediction of the secondary structure of the antimicrobial peptide TgHbpe

[0037] The secondary structure of the antimicrobial peptide TgHbpe was predicted by the online software APD2 and SOPMA, and it was found that the antimicrobial peptide may have an α-helical structure.

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PUM

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Abstract

The invention discloses an antimicrobial peptide TgHbpe and a mutant thereof. A section of antimicrobial peptide TgHbpe with an antibacterial property is separated from a hemoglobin subunit of tegillarca granosa, and site-specific mutagenesis is carried out on the antimicrobial peptide TgHbpe to obtain a mutant antimicrobial peptide mTgHbpe; Through functional analysis and identification, it is found that the two antimicrobial peptides have a relatively strong bactericidal effect on escherichia coli and vibrio alginolyticus, and the antimicrobial activity of the mutated and screened mTgHbpe isobviously higher than that of TgHbpe; the MIC of the antimicrobial peptide TgHbpe to escherichia coli is about 120 [mu] M, and the MIC of the antimicrobial peptide TgHbpe to vibrio alginolyticus is about 100 [mu] M; the MIC of the antimicrobial peptide mTgHbpe to escherichia coli is about 20 [mu] M, and the MIC of the antimicrobial peptide mTgHbpe to vibrio alginolyticus is about 7 [mu] M; and therefore, a foundation is laid for development and utilization of the hemoglobin source antimicrobial peptides of the tegillarca granosa.

Description

technical field [0001] The invention belongs to the technical field of applied marine organisms, and in particular relates to an antibacterial peptide TgHbpe and a mutant thereof. Background technique [0002] Shellfish is an important object of mariculture in my country, an important part of mariculture, and has an extremely important role in promoting my country's economy. With the continuous increase of people's demand for shellfish, shellfish farming has developed rapidly. Mud clam (Tegillarca granosa), belonging to the mollusk phylum, bivalves, order Dendonta, cockle family, cockle genus, is a traditional Chinese cultured shellfish. Mud clam meat is delicious, and the shell can be used as medicine, which has the effect of eliminating blood clots and reducing phlegm. Because of its affordable price, it is loved by people in many coastal cities. [0003] Hemoglobin (Hb) is a special protein that transports oxygen in red blood cells and makes blood red. All vertebrates ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/805A61K38/42A61P31/04
CPCC07K14/805A61P31/04A61K38/00Y02A50/30
Inventor 包永波郑晓莹张扬
Owner ZHEJIANG WANLI UNIV