Synthetic antibacterial peptide and preparation method and application thereof
An antibacterial peptide, chemical synthesis technology, applied in the preparation methods of peptides, antibacterial drugs, antifungal agents, etc., can solve the problems of low antibacterial activity and low clinical value, and achieve strong bactericidal activity and broad-spectrum killing activity. Effect
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Embodiment 1
[0059] Example 1 Preparation and separation and purification of antimicrobial peptides
[0060] In this example, the solid-phase chemical synthesis method was adopted, and the instrument used was the Pioneer polypeptide synthesizer produced by Applied Biosystems, Inc. of the United States. The synthesized peptide was sheared by high-concentration TFA, then purified by reverse column, and the purified peptide was identified by mass spectrometry. The specific test steps are as follows:
[0061] 1. Preparation of antimicrobial peptides (taking the preparation of 0.1 mmol amount as an example)
[0062] All the following reagents for the preparation of antimicrobial peptides were purchased from Applied Biosystems.
[0063] 1.1 Preparation of antimicrobial peptide BQ
[0064] The BQ polypeptide sequence is SEQ ID NO: 2 in the sequence listing:
[0065] BQ: Lys Trp Lys Ser Phe Ile Lys Lys Leu Thr Ser Lys Phe Leu His Ser Ala Lys Lys Lys Phe
[0066] The preparation of the antimic...
Embodiment 2
[0082] Example 2 Detection of Bactericidal Activity and Hemolytic Activity in Vitro of Synthetic Antimicrobial Peptides
[0083] The strains Pseudomonas aeruginosa CMCC10104 and Staphylococcus aureus CMCC26003 used in the following examples were purchased from China Institute for the Control of Biological Products.
[0084] The bactericidal activity of the synthetic antimicrobial peptides was detected by the 96-well plate method, and the antibacterial peptides cecropin A1 and buforin II synthesized by the solid-phase chemical method were used as controls to evaluate the bactericidal activity of various antimicrobial peptides in the present invention.
[0085] Measure the bactericidal activity of antimicrobial peptides according to the following steps:
[0086] Recover the strains, inoculate the slant and cultivate overnight at 37°C, pick the bacteria in ordinary LB medium, cultivate overnight at 37°C, dilute the bacterial solution to make the bacterial concentration 10 4 -10 ...
Embodiment 3
[0093] Example 3 Detection of Bactericidal Activity of Cyclic Derivatives of Antimicrobial Peptides
[0094] The modified products after cyclization of polypeptides such as BQ were synthesized by solid phase synthesis. For the synthesis method, see the operating guide for the pioneer peptide synthesizer of Applied Biosystems. After the synthetic product was separated and purified, the minimum inhibitory concentration of the derivative against Pseudomonas aeruginosa was tested, and the bactericidal activity was tested as in Example 2, and the test results are shown in Table 3.
[0095] Table 3 Detection of bactericidal activity of cyclized derivatives of antimicrobial peptides
[0096]
[0097]The results show that the antibacterial peptide of the present invention still maintains good antibacterial activity after cyclization.
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