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Litopenaeus vannamei haemocyanin antibacterial peptide and applications thereof

A technology of hemocyanin and antibacterial peptides, applied in the directions of antibacterial drugs, peptide/protein components, peptides, etc., can solve the problems of poor sustainable development prospects, low efficiency of shrimp breeding industry, and insufficient development stamina.

Inactive Publication Date: 2017-12-22
SHANTOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, compared with the international shrimp farming industry, the relative efficiency of the shrimp farming industry in our country and our province is relatively low, the development stamina is insufficient, and the prospect of sustainable development is not optimistic

Method used

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  • Litopenaeus vannamei haemocyanin antibacterial peptide and applications thereof
  • Litopenaeus vannamei haemocyanin antibacterial peptide and applications thereof
  • Litopenaeus vannamei haemocyanin antibacterial peptide and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Prediction of Litopenaeus vannamei hemocyanin antimicrobial peptide

[0019] First, use the three online prediction software of AntiBP Server, CAMP and APD2 to predict the large and small subunit antibacterial peptides of Litopenaeus vannamei hemocyanin. As shown in Table 1 and Table 2, a total of 34 predicted antibacterial activities are obtained. Small molecule fragments, of which 11 polypeptide sequences (in bold and bold) may form an α-helical structure, and can interact with pathogenic bacteria membranes to exert antibacterial effects. These predicted polypeptide molecular weights range from 1.5 to 1.9 kDa. In addition, the prediction results show that there are active fragments in the alpha helix domain at the N-terminus of hemocyanin, the Ig-like domain at the C-terminus, and the conserved copper ion binding domain in the middle part, and the copper ion binding domain and Ig-like domain occupy the main area.

[0020] Table 1 Prediction of antimicrobial pepti...

Embodiment 2

[0028] Example 2 Analysis of the secondary structure content of Litopenaeus vannamei hemocyanin antimicrobial peptide by circular dichroism

[0029] The 11 kinds of peptides described in implementation 1 (the bolded peptides in Table 1 and Table 2) that may form an α-helical structure and can interact with the pathogen membrane to play a bacteriostatic effect are scanned and analyzed by circular dichroism Its structural content.

[0030] 2.1 Experimental equipment

[0031] 2.1.1 Experimental materials

[0032] The antibacterial peptides used in the experiment were synthesized by SciLight Biotechnology, LLC (SciLight Biotechnology, LLC). The peptides were N-terminally acetylated and C-terminally amidated, with a purity of ﹥95%.

[0033] 2.1.2 Experimental instrument

[0034] Ultraviolet spectrophotometer (UV5200) manufacturer: Shanghai Yuanxi Instrument Co., Ltd.

[0035] Circular dichrograph (Jasco J-810) manufacturer: Japan Jasco

[0036] Cuvette: 1mm quartz cuvette

[0037] 2.2 Sample pr...

Embodiment 3

[0073] Example 3 Using plate counting method to verify the antibacterial activity of antimicrobial peptide B14

[0074] 3.1 Operation steps

[0075] The antibacterial peptides used in the experiment were synthesized by SciLight Biotechnology, LLC (SciLight Biotechnology, LLC). The peptides were N-terminally acetylated and C-terminally amidated, with a purity of ﹥95%.

[0076] The bacteriostatic activity analysis of the chemically synthesized antimicrobial peptides from shrimp hemocyanin was carried out by the following method, and the specific process is as follows:

[0077] (1) First, activate a small amount of pathogenic bacteria (Vibrio parahaemolyticus, Streptococcus beta, Escherichia coli, Vibrio alginolyticus, Aeromonas hydrophila, Vibrio river) stored in -80℃ (1mL broth liquid medium + 50μL strain), 37°C constant temperature shaker culture for 6-12h to logarithmic growth phase (or about 24h at 28°C).

[0078] (2) According to the original concentration of the bacterial solution,...

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Abstract

The present invention discloses a litopenaeus vannamei haemocyanin antibacterial peptide, which has the amino acid sequence represented by SEQ ID NO:1 in the sequence table, and has the molecular weight of 1875 Da. The invention further discloses applications of the litopenaeus vannamei haemocyanin antibacterial peptide, particularly applications of the litopenaeus vannamei haemocyanin antibacterial peptide in preparation of drugs for treatment or prevention of Gram-negative bacteria. According to the present invention, the foundation is established for the subsequent further study on the immunologic defence mechanism of litopenaeus vannamei and the development of antibacterial drugs.

Description

Technical field [0001] The invention relates to the technical field of aquaculture, in particular to a Litopenaeus vannamei hemocyanin antibacterial peptide and its application. Background technique [0002] In recent years, with the development of the economy, people's demand for aquatic products has increased day by day. When the catch cannot meet the market demand, the aquaculture industry has developed rapidly. Shrimp is one of the fastest growing products in aquaculture, and my country is the country with the highest annual production of prawns. However, compared with the international shrimp aquaculture industry, the relative efficiency of the shrimp aquaculture industry in our country and our province is relatively low, the development potential is insufficient, and the prospects for sustainable development are not optimistic. The reason is mainly related to the frequent occurrence of diseases in shrimp farming in my country and the repeated trade barriers in shrimp expor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435A61K38/17A61P31/04
CPCA61K38/00C07K14/43509Y02A50/30
Inventor 章跃陵杨燊黄河钟名其王帆
Owner SHANTOU UNIV
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