Producing method of antimicrobial peptide with enhanced adhesion and uses thereof
An antimicrobial peptide and antimicrobial technology, applied in botany equipment and methods, chemicals for biological control, medical preparations of non-effective ingredients, etc., can solve the problem of chemical toxicity, silver and copper do not have antimicrobial properties Activity, complex immobilization methods and other issues, to achieve the effect of no cytotoxicity and excellent antimicrobial activity
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Embodiment 1
[0059] Example 1: Preparation of Adhesive Antimicrobial Peptides
Embodiment 1-1
[0060] Example 1-1: Adhesiveness imparted to antimicrobial peptides
[0061] In order to utilize NKC, a peptide with strong antibacterial activity against various microorganisms, as an antimicrobial coating agent, the antimicrobial peptide was endowed with adhesion by conjugating 3,4-dihydroxyphenylalanine (DOPA) to the end of NKC properties, the aforementioned DOPA is known as an unnatural amino acid and plays an important role in providing the adhesion strength of mussel adhesion protein, which is biocompatible and exhibits strong adhesion regardless of the surface sex. In this paper, to determine the optimal number of DOPA, 1, 3, 5 or 7 DOPA moieties were conjugated to the end of NKC, and then GGGGS was added as a flexible linker sequence between NKC and DOPA sequences to prevent NKC and DOPA affect each other's activities. Amino acid sequence information for conjugated NKC and DOPA is shown in Table 1 below.
[0062] Table 1
[0063]
[0064] *DOPA is abbreviated as...
Embodiment 1-2
[0065] Examples 1-2: Identification of antimicrobial activity of adhesive antimicrobial peptides
[0066] In order to confirm the antimicrobial activity of the adhesive antimicrobial peptide prepared in Experimental Example 1-1, the minimum inhibitory concentration for inhibiting the growth of Escherichia coli was identified by using the method disclosed in Nature Protocols.2008; 3(2):163-175 (MIC). Briefly, media in which individual colonies were grown for 16 h was subcultured and grown until OD 600 0.4 to 0.6 is reached, which corresponds to logarithmic growth. Thereafter, calculate the cell number based on the OD value, and set the cell number as 10 6 cells / mL. When loading M-H Broth (MHB) into the polypropylene 96-well plate, add 90 µL to the first row of the plate and 100 µL to the control wells and 50 µL to the remaining wells. Thereafter, prepare a peptide solution with a concentration of 640 μM and add 10 μL of it to each well of the first row. Mix the peptide sol...
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