Antibacterial peptide and use thereof

Abstract

The invention relates to the field of molecular microbiology, in particular to antibacterial peptide and use thereof. The antibacterial peptide T1-17 is represented by an amino acid sequence SEQ ID No.1 in a sequence table. The antibacterial peptide T1-17 has a bacterial inhibition function. The antibacterial peptide has a high-efficiency bacterial inhibition function and can effectively inhibit various kinds of fish pathogenic bacteria. Thus, the antibacterial peptide is expected to be used for preventing and controlling fish diseases.

Description

technical field

[0001] The invention relates to the field of molecular microbiology, in particular to an antibacterial peptide and its application. Background technique

[0002] With the continuous improvement of fishery intensive farming, the ecological environment of the water area has been seriously damaged, resulting in an increasing number of aquatic animal diseases, and the disease problem has become a bottleneck for the sustainable development of aquaculture. Traditional antibiotic treatment not only leads to the emergence of bacterial resistance, but also causes drug residues, which seriously affects the quality of aquatic products and even threatens human health. Therefore, it is very urgent to find new disease-resistant substances to replace antibiotics. Antimicrobial peptides are a class of small molecule polypeptides that widely exist in microorganisms, animals and plants. They are usually composed of more than 30 amino acid residues and have a molecular weight ...

Examples

Embodiment 1

[0012] The antimicrobial peptide T1-17 of the present invention has the amino acid sequence in SEQ ID No.1 of the sequence listing.

[0013] The sequence listing SEQ ID No.1 is:

[0014] KCRRRKVHGPMIRIRKK

[0015] (a) Sequence features:

[0016] ●Length: 17

[0017] ●Type: amino acid sequence

[0018] ●Chain type: single chain

[0019] ●Topological structure: linear

[0020] (b) Molecule type: protein

[0021] (c) Assumption: No

[0022] (d) Antisense: no

[0023] (e) Original source: Synthetic

[0024] The antimicrobial peptide T1-17 synthesized in this example was synthesized according to the BOC solid-phase peptide synthesis method, and the antimicrobial peptide T1-17 with a purity greater than 90% was finally obtained. During the synthesis, a Boc-amino acid derivative is covalently cross-linked to the resin (such as an insoluble polymer resin, namely chloromethyl polymer resin), the Boc is removed with TFA, and the free amino terminal is neutralized with triethyla...

Embodiment 2

[0027] Application of Antimicrobial Peptide T1-17

[0028] 1) Preparation of Edwardsiella tarda and Vibrio anguillarum. Cultivate Edwardsiella tarda and Vibrio anguillaris to OD in LB medium 600 0.5, and then centrifuged (5000g, 4°C) for 10min. Collect the cells and suspend them in PBS to a final concentration of 1x10 6 cfu / ml.

[0029] The PBS composition is by weight percentage: 0.8% NaCl, 0.02% KCl, 0.358% Na 2 HPO 4 .12H 2 O, 0.024% NaH 2 PO 4 .

[0030] 2) Bactericidal effect of antimicrobial peptide T1-17. Mix 10 ul of the bacteria solution in step 1) above with 30 nMT1-17 or PBS (control), and incubate at 25° C. for 3-5 hours. Then the mixture was spread on LB solid plates and cultured at 28°C for 32-48h. Colony counts found that compared with PBS, T1-17 treatment resulted in a reduction of about 90% of the number of Edwardsiella tarda, and Vibrio anguillaris was completely non-viable, that is, about 90% of Edwardsiella tarda and 100% of Vibrio anguillarum we...

technical field

[0001] The invention relates to the field of molecular microbiology, in particular to an antibacterial peptide and its application. Background technique

[0002] With the continuous improvement of fishery intensive farming, the ecological environment of the water area has been seriously damaged, resulting in an increasing number of aquatic animal diseases, and the disease problem has become a bottleneck for the sustainable development of aquaculture. Traditional antibiotic treatment not only leads to the emergence of bacterial resistance, but also causes drug residues, which seriously affects the quality of aquatic products and even threatens human health. Therefore, it is very urgent to find new disease-resistant substances to replace antibiotics. Antimicrobial peptides are a class of small molecule polypeptides that widely exist in microorganisms, animals and plants. They are usually composed of more than 30 amino acid residues and have a molecular weight ...

Examples

Embodiment 1

[0012] The antimicrobial peptide T1-17 of the present invention has the amino acid sequence in SEQ ID No.1 of the sequence listing.

[0013] The sequence listing SEQ ID No.1 is:

[0014] KCRRRKVHGPMIRIRKK

[0015] (a) Sequence features:

[0016] ●Length: 17

[0017] ●Type: amino acid sequence

[0018] ●Chain type: single chain

[0019] ●Topological structure: linear

[0020] (b) Molecule type: protein

[0021] (c) Assumption: No

[0022] (d) Antisense: no

[0023] (e) Original source: Synthetic

[0024] The antimicrobial peptide T1-17 synthesized in this example was synthesized according to the BOC solid-phase peptide synthesis method, and the antimicrobial peptide T1-17 with a purity greater than 90% was finally obtained. During the synthesis, a Boc-amino acid derivative is covalently cross-linked to the resin (such as an insoluble polymer resin, namely chloromethyl polymer resin), the Boc is removed with TFA, and the free amino terminal is neutralized with triethyla...

Embodiment 2

[0027] Application of Antimicrobial Peptide T1-17

[0028] 1) Preparation of Edwardsiella tarda and Vibrio anguillarum. Cultivate Edwardsiella tarda and Vibrio anguillaris to OD in LB medium 600 0.5, and then centrifuged (5000g, 4°C) for 10min. Collect the cells and suspend them in PBS to a final concentration of 1x10 6 cfu / ml.

[0029] The PBS composition is by weight percentage: 0.8% NaCl, 0.02% KCl, 0.358% Na 2 HPO 4 .12H 2 O, 0.024% NaH 2 PO 4 .

[0030] 2) Bactericidal effect of antimicrobial peptide T1-17. Mix 10 ul of the bacteria solution in step 1) above with 30 nMT1-17 or PBS (control), and incubate at 25° C. for 3-5 hours. Then the mixture was spread on LB solid plates and cultured at 28°C for 32-48h. Colony counts found that compared with PBS, T1-17 treatment resulted in a reduction of about 90% of the number of Edwardsiella tarda, and Vibrio anguillaris was completely non-viable, that is, about 90% of Edwardsiella tarda and 100% of Vibrio anguillarum we...

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