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

A technology of immunomodulatory activity 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: 2021-12-17
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 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, anti-inflammatory, endotoxin neutralizing and immunomodulatory activities, its preparation method and application
  • A multifunctional hybrid peptide with antibacterial, anti-inflammatory, endotoxin neutralizing and immunomodulatory activities, its preparation method and application
  • A multifunctional hybrid peptide with antibacterial, anti-inflammatory, endotoxin neutralizing and immunomodulatory activities, its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Obtaining of Multifunctional Hybrid Peptide CLP

[0056] By screening the polypeptides with immune regulation, anti-inflammatory and antibacterial functions, it is determined to immune anti-inflammatory polypeptide LTP (sequence shown in SEQ ID NO.2) and antibacterial and antiviral polypeptide CL (sequence shown in SEQ ID NO.1 ) as the parent peptide. The sequence, structure, and relationship between sequence structure and function of LTP and CL were studied, and multiple candidate hybrid peptides were obtained by using protein molecular design technology to hybridize polypeptides LTP and CL. Among them, CLP has the best immunomodulatory, anti-inflammatory, antibacterial and endotoxin-neutralizing activities, and CLP includes the amino acid sequence shown in SEQ ID NO.1 and the 19th to 29th amino acid sequence in SEQ ID NO.2, The specific amino acid sequence is shown in SEQ ID NO.3. In this embodiment, the hybrid peptides CLP (sequence shown in SEQ ID NO.3),...

Embodiment 2

[0059] The antibacterial effect of embodiment 2 multifunctional hybrid peptide CLP

[0060] The minimum inhibitory concentration (MIC) of multifunctional hybrid peptide CLP 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 CLP 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 CL.

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

[0062]

Embodiment 3

[0063] Example 3 The neutralization effect of hybrid peptide CLP on LPS

[0064] Dissolve and dilute the hybrid peptide CLP and its parent peptides LTP and CL into solutions of different concentrations (0-64 μg / mL) with pyrogen-free endotoxin test water, and take 100 μL of the above-mentioned peptide solutions of each concentration and LPS (1EU / mL) respectively. mL) mixed. After incubation at 37°C for 30 min, the neutralization rate of polypeptides CLP, LTP, and CL to LPS was detected using a chromogenic mechanism Limulus kit, and polymyxin B (PMB) was used as a control. The result is as figure 2 As shown, the polypeptide CLP has a high LPS neutralizing activity, and its LPS neutralizing activity is equivalent to that of polymyxin B. When the concentration is 4 μg / mL, the neutralization rate of the hybrid peptide CLP to LPS is close to 100%, and The neutralizing activity was significantly higher than that of its parent peptides LTP and CL.

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Abstract

The invention relates to the technical field of protein engineering, in particular to a multifunctional hybrid peptide with antibacterial, anti-inflammatory, endotoxin neutralizing and immunoregulatory activities, as well as 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 the 19th sequence of SEQ ID NO.2 ~ 29 shown. The hybrid peptide has the functions of antibacterial, endotoxin neutralization, immune regulation and anti-inflammation, and has the advantages of low cytotoxicity, high safety, convenient preparation and low cost, and can be used as an ideal antibacterial agent and endotoxin neutralizer , immunomodulator and anti-inflammatory agent, widely used in human and animal medicine, food, health care, feed, nutrition and other fields, with good application potential and value.

Description

technical field [0001] The invention relates to the technical field of protein engineering, in particular to a hybrid peptide with antibacterial, anti-inflammatory, endotoxin-neutralizing and immune-modulating activities, 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 (such as endotoxin) and other metabolites, which can induce inflammation and oxidative damage, etc., and damage the immune system in severe cases. 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62A61K38/16A61P31/04A61P31/00A61P31/12A61P29/00A61P37/02A23L33/18A23K20/147
CPCC07K14/00A61P31/04A61P31/00A61P31/12A61P29/00A61P37/02A23L33/18A23K20/147C07K2319/00A61K38/00A23V2002/00A23V2200/324A23V2200/30A23V2250/55
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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