Human alpha-defensin-5 modifier and applications thereof
A defensin, amino acid technology, applied in animal/human peptides, animal/human proteins, medical preparations containing active ingredients, etc., can solve the problems of reducing antibacterial activity, the impact of antibacterial activity, etc. effect, the effect of improving salt tolerance
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Embodiment 1
[0037] The amino acid sequence of HD-5 is shown in SEQ ID NO.1, specifically:
[0038] Ala 1 -Thr 2 -Cys 3 -Tyr 4 -Cys 5 -Arg 6 -Thr 7 -Gly 8 -Arg 9 -Cys 10 -Ala 11 -Thr 12 -Arg 13 -Glu 14 -Ser 15 -Leu 16 -Ser 17 -Gly 18 -Val 19 -Cys 20 -Glu 21 -Ile 22 -Ser 23 -Gly 24 -Arg 25 -Leu 26 -Tyr 27 -Arg 28 -Leu 29 -Cys 30 -Cys 31 -Arg 32 , where Cys 3 and Cys 31 、Cys 5 and Cys 20 、Cys 10 and Cys 30 Intramolecular disulfide bonds are formed between them.
[0039] The modified HD-5Myr and HD-5Lau of the present invention are obtained by modifying the C-terminus of the HD-5 molecule. The amino acid sequences of HD-5Myr and HD-5Lau are shown in SEQ ID NO.2.
[0040] HD-5Myr consists of 34 amino acid residues with a molecular weight of 3976.8Da.
[0041] The amino acid sequence of HD-5Myr is:
[0042] Ala 1 -Thr 2 -Cys 3 -Tyr 4 -Cys 5 -Arg 6 -Thr 7 -Gly 8 -Arg 9 -Cys 10 -Ala 11 -Thr 12 -Arg 13 -Glu 14 -Ser 15 -Leu 16 -Ser 17 -Gly 1...
Embodiment 2
[0050] The HD-5 and HD-5Myr in the above-mentioned Example 1 were synthesized by "Shanghai Gill Polypeptide Co., Ltd.". The HD-5Lau and MyrHD-5 of the above-mentioned Example 1 were synthesized by "Hangzhou Zhongpei Biochemical Co., Ltd.".
Embodiment 3
[0052] The antibacterial activity of the antimicrobial peptide was measured, and the strength of the antibacterial effect of the HD-5 modified body HD-5Myr, HD-5Lau and MyrHD-5 synthesized in Example 2 was compared with that of the natural HD-5.
[0053] The experiment was carried out according to the method for detecting antibacterial peptide antibacterial activity reported by Ericksen et al. in 2005 (Ericksen B, et al., Antibacterial activity and specificity of the six human{alpha}-defensins. Antimicrob Agents Chemother, 2005).
[0054] Bacteria were inoculated on LB agar plates and cultured at a constant temperature of 37°C until colonies grew. Pick a single colony, inoculate it into 5ml LB medium, and incubate at a constant temperature of 37°C for 4-6 hours with shaking. Wash 2 times with sterile water. Measure the absorbance at 600nm on a multifunctional microplate reader, 1OD 600 =4×10 8 CFU / ml, the test bacteria were diluted to 2×10 with sterile water 6 CFU / ml.
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