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Antimicrobial peptide HP10, preparing method thereof and application thereof

An antibacterial peptide and a synthesis method are applied in the field of antibacterial peptide HP10 and its preparation, which can solve the problems of restricting the popularization and application of antibacterial peptides, toxic and side effects of animal cells, and low antibacterial activity, so as to enhance the immunity of the body, improve the growth performance of piglets, Strong bactericidal effect

Active Publication Date: 2015-12-16
GUANGZHOU AONONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the antimicrobial peptides from natural sources have defects such as large molecular weight, immunogenicity, low antibacterial activity, toxic side effects on animal cells, or hemolysis, which to a certain extent limits the popularization and application of antimicrobial peptides in pig breeding

Method used

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  • Antimicrobial peptide HP10, preparing method thereof and application thereof
  • Antimicrobial peptide HP10, preparing method thereof and application thereof
  • Antimicrobial peptide HP10, preparing method thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Preparation of antimicrobial peptide HP10:

[0021] The antibacterial peptide HP10 is intercepted and modified on the basis of the natural antibacterial peptide PMAP36 (GRFRRLRKKTRKRLKKIGKVLKWIPPIVGSIPLGCG). Since histidine is an essential amino acid for juvenile animals, and its antibacterial activity is stronger than that of arginine and lysine, so Replace arginine and lysine with histidine.

[0022] The chemical synthesis method of antimicrobial peptide HP10 comprises the following steps:

[0023] (1) According to the amino acid sequence HHHLHHIHKP of the antimicrobial peptide HP10, the antimicrobial peptide HP10 was activated and cross-linked one by one from the C-terminal to the N-terminal by the FMOC protection synthesis method, which was synthesized by Shanghai Gil (GL) Biochemical Co., Ltd. Synthesize the full sequence of antimicrobial peptides through a fully automatic peptide synthesizer (ABI433);

[0024] (2), the product of step (1) is desalte...

experiment example 1

[0029] Experimental example 1: Antibacterial peptide HP10 antibacterial test:

[0030] (1), Test strains: Escherichia coli, Salmonella, Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis and Bacillus megaterium.

[0031] (2), polypeptide mother solution (0.2% BSA+0.01% acetic acid): Weigh 0.2g bovine serum albumin (BSA) and add 80mLddH 2 Mix well in O water, then add 10 μL of acetic acid solution, then wash with ddHO 2 Dilute to 100 mL with O water, filter and sterilize with a filter membrane with a pore size of 0.22 μm, and store at 4°C for later use.

[0032] The antibacterial effect was reflected by measuring the minimum inhibitory concentration (MIC) of the antimicrobial peptide HP10, which was the lowest sample concentration at which no bacterial growth could be detected. Cultivate the test bacteria overnight at 37°C for 16 hours, then transfer 100 μL of the overnight cultured bacterial solution to fresh MHB broth medium, and cultivate for 2.5 to 3 h...

experiment example 2

[0036] Experimental example 2: Effects of antimicrobial peptide HP10 on growth performance, intestinal health and immune function of weaned piglets:

[0037] A total of 320 weaned piglets at the age of 21 days were selected, and were randomly divided into 4 treatment groups according to body weight and sex according to the experimental design scheme shown in Table 2. The negative control group was the basal diet, and the HP10- The 50 group was added 50mg / kg antimicrobial peptide HP10 to the basic diet; 10 repetitions, 8 pigs per repetition, 14 days of test period.

[0038] Table 2 Experimental Design Scheme

[0039]

[0040] The effects of antimicrobial peptide HP10 on production performance, diarrhea rate and immunoglobulin of weaned piglets are shown in Table 3, figure 1 and Table 4. As can be seen from the experimental results in Table 3, the antimicrobial peptide HP10 can significantly improve the growth performance of weaned piglets; figure 1 It can be seen from ...

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Abstract

The invention discloses antimicrobial peptide HP10, a preparing method thereof and an application thereof, and belongs to the technical field of synthesis of peptide with 5-20 pieces of amino acid and the technical field of specific curative activity of compounds and medicinal preparations. The amino acid sequence of the antimicrobial peptide HP10 is HHHLHHIHKP, the molecular weight of the antimicrobial peptide HP10 is 1,292.4 Da, and the isoelectric point of the antimicrobial peptide HP10 is 8.79. The preparing method of the antimicrobial peptide HP10 includes the following steps that 1, the antimicrobial peptide HP10 with the amino acid sequence of HHHLHHIHKP is activated and crosslinked from the C end to the N end with an FMOC protection synthesis method; 2, chromatography, desalination and purification are carried out on the product of the step 1 through an HPLC reversed-phase column, and the antimicrobial peptide HP10 is obtained. The antimicrobial peptide HP10 has the advantages that the molecular weight is small, artificial synthesis is convenient, heat stability is high, the sterilizing effect is high, and drug resistance is avoided; the antimicrobial peptide HP10 further has the advantages that when the antimicrobial peptide HP10 is applied to piglet feed, the intestinal health of piglets can be remarkably improved, body immunity can be improved, and the growth performance of the piglets can be improved.

Description

technical field [0001] The invention belongs to the technical field of peptide synthesis containing 5-20 amino acids and the technical field of specific therapeutic activity of compounds and pharmaceutical preparations, and specifically relates to antibacterial peptide HP10 and its preparation method and application. Background technique [0002] Feed antibiotics have played a vital role in the development of the global livestock industry. In recent years, in order to pursue greater economic benefits, the abuse of antibiotics in animal production has become increasingly serious, which has not only caused the problem of veterinary drug residues in animal foods, but also led to the emergence of drug-resistant strains, which has threatened the sustainable development of animal husbandry. caused serious trouble. Therefore, developed countries such as the European Union and South Korea have completely banned the use of antibiotics in livestock and poultry feed, and finding alter...

Claims

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Application Information

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IPC IPC(8): C07K7/06A23K1/16A23K1/18C07K1/20C07K1/10C07K1/06
Inventor 姜文周盛昌苏斌朝施建成肖丽萍叶俊标
Owner GUANGZHOU AONONG BIOTECH
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