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Antibacterial peptide DC and its preparing process and use

A technology of antimicrobial peptides and engineering bacteria, applied in the field of antimicrobial peptides, which can solve the problems of high cost, unsuitable industrial production, low extraction yield, and low content of natural antimicrobial peptides

Inactive Publication Date: 2007-06-20
GUANGDONG COOWAY BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the low content of natural antimicrobial peptides, the extraction rate is low (0.005%), and the cost is high, so it cannot be industrialized.

Method used

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  • Antibacterial peptide DC and its preparing process and use
  • Antibacterial peptide DC and its preparing process and use
  • Antibacterial peptide DC and its preparing process and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] 1. Design and synthesis of antimicrobial peptide DC gene

[0101] According to the structure of the bactericidal activity of natural antimicrobial peptides, the antimicrobial peptide DC gene was designed and synthesized. The gene is characterized by two α-helices, the 23rd amino acid in the first position constitutes the first hydrophilic helix, and the 27th to 39th amino acids constitute the second hydrophobic helix. Between two α-helices, there is an A-G-P (alanine-glycine-proline) connection, forming a β-sheet. The 39th amino acid residue at the C-terminus is asparagine (N), which is similar to the lysinamide at the end of the natural antimicrobial peptide.

[0102] In the present invention, the designed antimicrobial peptide DC gene is synthesized with a DNA synthesizer for the main chain and auxiliary chain at one time, and annealed to form a 144bp antimicrobial peptide DC gene, in which 120bp of the antimicrobial peptide DC is completely consistent with the desig...

Embodiment 2

[0112] Embodiment 2 antimicrobial peptide DC application effect

[0113] 1. The effect of antimicrobial peptides in the treatment of drug-resistant Pseudomonas aeruginosa on upper respiratory tract infection

[0114] The hospital admitted 5 patients with upper respiratory tract infection, complaining of pharyngitis, cough, and no fever. Pseudomonas aeruginosa was isolated from the pharynx and cough sputum, which was Gram-negative.

[0115] Antibiotic resistance testing of isolated Pseudomonas aeruginosa. The bacteria isolated from 3 of the patients were resistant to the following antibiotics. The results are shown in Table 5:

[0116] Table 5 Resistance test of drug-resistant Pseudomonas aeruginosa to various antibiotics

[0117] antibiotics

[0118] Decided to use antimicrobial peptide preparation 2500 units / ml for spray treatment. Use SY-II type nebulizer, temperature 60 ℃, treat antimicrobial peptide liquid as oral spray treatment, about 100ml each time, three...

Embodiment 3

[0127] The safety evaluation of embodiment 3 antimicrobial peptides

[0128] The bactericidal activity of antibacterial peptide DC fermentation broth purified by ion exchange was 50,000 units / g. Acute and long-term toxicity tests and "three-to-one" tests are carried out respectively to confirm its safety.

[0129] 1. Acute toxicity test

[0130] Experimental animals 50 white mice of insect species were administered orally according to different doses of antimicrobial peptides, and the results of poisoning within one week of investigation are shown in Table 7.

[0131] Table 7 Antimicrobial Peptides Oral Acute Toxicity Test to Mice (7 days)

[0132] Dose (g / kg)

[0133] Tests have shown that antimicrobial peptides are not toxic to animals during oral administration, and the lethal dose (LD50) cannot be detected. The maximum tolerated dose is 5g / kg level.

[0134] 2. Antimicrobial peptide long-term toxicity test

[0135] Experimental animals 60 white rats were ran...

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Abstract

The invention discloses an antibiotic peptide and its preparation and application. According to sequence of natural antibiotic peptide, gene of antibiotic peptide is designed and synthesized in DNA synthesizer by choosing genetic code preferred by yeast. The invention selects dispense of culture medium and optimizes condition of fermentation. The antibiotic peptide can kill many pathogenic microorganisms such as Gram-positive bacteria, Gram-negative bacterium and so on.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an antibacterial peptide. Background technique [0002] Cecropin is a bactericidal polypeptide induced by injecting Escherichia coli into tussah silkworm pupae. In 1981, Huang Ziran et al. first reported the isolation of antibacterial peptides from tussah silkworm, which has multiple members such as B and D, and has a broad-spectrum bactericidal function. Natural antimicrobial peptides can be used as raw materials for medicines. For example, in 1996, it was used as a new drug "Shenganning" for the treatment of nephritis and hepatitis. The approval number is Yuewei Yaozhunzi (1996) No. 107388, and it has been put into production. In 1997, tussahin (containing antimicrobial peptide) capsules as a second-class western medicine for the treatment of hepatitis B has entered into clinical observation, with the approval number (97) XL-29. Due to the low content of natural antimicrobial p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/63C12P21/02A61K38/17A61P11/04A61P11/02A61P15/02A61P31/04
Inventor 黄自然陈松彬黄永彤
Owner GUANGDONG COOWAY BIOTECH CO LTD
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