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A multifunctional hybrid peptide with antibacterial, antiviral, immunomodulatory and anti-inflammatory activities and its preparation method and application

A technology of immune regulation and hybrid peptide, applied in the field of protein engineering, can solve the problems of high cytotoxicity, weak anti-inflammatory ability, weak antibacterial activity, etc., and achieve the effect of low cytotoxicity, convenient preparation and low cost

Active Publication Date: 2021-11-09
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, natural bioactive peptides also have some defects, mainly as follows: (1) Some natural bioactive peptides have relatively low biological activity and single function, such as the problem of weak antibacterial activity of pig lactoferrin peptide ; and although cecropin A has good antibacterial efficacy, its anti-inflammatory ability is relatively weak; (2) Some natural bioactive peptides have high cytotoxicity, which can cause serious damage to the body while inhibiting and killing pathogenic microorganisms (such as melittin and LL-37)

Method used

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  • A multifunctional hybrid peptide with antibacterial, antiviral, immunomodulatory and anti-inflammatory activities and its preparation method and application
  • A multifunctional hybrid peptide with antibacterial, antiviral, immunomodulatory and anti-inflammatory activities and its preparation method and application
  • A multifunctional hybrid peptide with antibacterial, antiviral, immunomodulatory and anti-inflammatory activities and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Preparation of Multifunctional Hybrid Peptide LPML

[0056] By screening polypeptides with immune regulation, anti-inflammatory, antibacterial and antiviral functions, it is determined to immune anti-inflammatory polypeptide LTP (sequence shown in SEQ ID NO.1) and antibacterial and antiviral polypeptide ML (sequence shown in SEQ ID NO. 2) as the parent peptide. The sequence, structure, and relationship between sequence structure and function of LTP and ML were studied, and multiple candidate hybrid peptides were obtained by using protein molecular design technology to hybridize polypeptides LTP and ML. Among them, LPML has the best antibacterial, antiviral, immunomodulatory and anti-inflammatory activities. LPML comprises the amino acid sequence shown in SEQ ID NO.1 and the amino acid sequence shown in SEQ ID NO.2, and the specific amino acid sequence is shown in SEQ ID NO.3. In this embodiment, the hybrid peptides LPML (sequence shown in SEQ ID NO.3), MLP (...

Embodiment 2

[0059] Antibacterial effect of embodiment 2 multifunctional hybrid peptide LPML

[0060] The minimum inhibitory concentration (MIC) of multifunctional hybrid peptide LPML on E.coil CVCC 245, E.coil K88, EHECO157:H7 and Salmonella CVCC 2212 was detected by microdilution method. The results are shown in Table 2. The results showed that the multifunctional hybrid peptide LPML had good antibacterial activity against the four tested Gram-negative bacteria, and its antibacterial activity was significantly higher than that of its parent peptides LTP and ML.

[0061] The minimum inhibitory concentration (MIC) of table 2 multifunctional hybrid peptide

[0062]

Embodiment 3

[0063] Example 3 Antiviral effect of multifunctional hybrid peptide LPML

[0064] VSV, IBV, and ARV were used as indicator viruses to detect the antiviral activity of the polypeptide. The Vero cells were seeded in 96-well cell culture plates at 3×105 cells / mL, and cultured overnight in a 5% CO2 incubator at 37°C to allow the cells to adhere to the wall. Remove the cell culture medium, and inoculate the virus liquid after 10-fold gradient dilution with the culture medium containing the final concentration of 10 μg / mL polypeptide in the cell culture plate, add 100 μL to each well, inoculate 8 wells for each dilution, blank control Add 10-fold gradient dilution of virus solution to the group, and culture in a 37°C carbon dioxide incubator for 3 to 5 days until the cells have no increase in new lesion holes. Use the Reed-Muench method to calculate its influence on the virus TCID50, and the calculation formula is as follows:

[0065] lgTCID50 = difference between distance ratio x ...

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Abstract

The invention relates to the technical field of protein engineering, in particular to a multifunctional hybrid peptide with antibacterial, antiviral, immune regulation and anti-inflammatory activities and its preparation method and application. The hybrid peptide provided by the present invention comprises: peptide I and peptide II fused with peptide I, wherein the amino acid sequence of peptide I is shown in SEQ ID NO.1, and the amino acid sequence of peptide II is shown in SEQ ID NO.2. The hybrid peptide has antibacterial, antiviral, immune regulation and anti-inflammatory functions, and has the advantages of low cytotoxicity, high safety, convenient preparation and low cost, and can be used as an ideal antibacterial agent, antiviral agent and immunomodulator And anti-inflammatory agent, widely used in human and animal medicine, food, health care, feed, nutrition and other fields, has good application potential and value.

Description

technical field [0001] The invention relates to the technical field of protein engineering, in particular to a multifunctional hybrid peptide with antibacterial, antiviral, immune regulation and anti-inflammatory activities and its preparation method and application. Background technique [0002] Bacterial infection is an infectious disease caused by pathogenic bacteria or conditional pathogenic bacteria invading the animal body to grow and reproduce, producing toxins and other metabolites, which can induce inflammation and oxidative damage, etc., and in severe cases damage the immune system and even lead to death. When animals are young, weak, sick, stressed, or infected with pathogens, their immunity is often reduced, and then secondary infections (bacterial infections, etc.) occur, causing inflammatory reactions. For the treatment of infection and inflammation, the traditional control strategy is to use antibiotics and chemical drug agents. The continuous use of antibiot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62A61K38/16A61P31/04A61P31/14A61P37/02A61P29/00A23L33/18A23K20/147
CPCA23V2002/00A61K38/00A23K20/147A23L33/18A61P29/00A61P31/04A61P31/14A61P37/02C07K14/00C07K2319/00A23V2200/324A23V2200/30A23V2250/55
Inventor 张日俊张璐璐卫旭彪黄燕斯大勇王晓冰郭贺楠
Owner CHINA AGRI UNIV
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