A kind of anti-candida albicans polypeptide and its application
A technology of Candida albicans and antibacterial drugs, applied in the field of biomedicine, can solve the problems of increased treatment difficulty and increased drug resistance rate of Candida albicans, and achieve rapid and efficient killing, good antibacterial activity, and high-efficiency killing effects
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Embodiment 1
[0021] The acquisition of anti-Candida albicans polypeptide (hereinafter referred to as polypeptide)
[0022] The anti-Candida albicans polypeptide proposed by the present invention is derived from the Chinese medicine slug polypeptide sequence database constructed by us, and its amino acid sequence is shown in SEQ ID No.1;
[0023] SEQ ID NO.1: Arg-Gly-Asp-Ile-Leu-Lys-Arg-Trp-Ala-Gly-His-Phe-Ser-Lys-Leu-Leu.
[0024] According to the amino acid sequence shown, it was synthesized by Jill Biochemical (Shanghai) Co., Ltd. by solid-phase chemical synthesis, purified by reverse-phase high-performance liquid chromatography and identified by electrospray mass spectrometry, and a pure peptide with a purity greater than 95% was obtained.
Embodiment 2
[0026] Antibacterial Activity of Anti-Candida Albicans Peptides
[0027] Prepare the peptide preservation solution with physiological salt as the solvent, and perform two-fold gradient dilution on the peptide preservation solution with physiological salt. Adjust the bacterial solution to be tested in the logarithmic growth phase to about 10 with fresh YPD medium 3 -10 4 cfu / mL. In a sterile 96-well cell culture plate, the above-mentioned diluted polypeptide solution and diluted bacterial solution were added to each well, and 3 parallels were performed for each polypeptide concentration. Incubate at a constant temperature of 35°C for 24 hours. The minimum inhibitory concentration (MIC) of the polypeptide to the tested bacteria is defined as the lowest concentration of the polypeptide at which there is no visible growth of Candida albicans with the naked eye. At the same time, the polypeptide concentration wells without visible Candida albicans growth were sampled and coated ...
Embodiment 3
[0032] Bactericidal effect of anti-Candida albicans polypeptide
[0033] Adjust the concentration of Candida albicans AY93025 bacteria in the logarithmic growth phase to about 10 with fresh YPD medium 4 -10 5 cfu / mL, the final concentrations of antimicrobial peptides added were 100 μg / mL and 200 μg / mL, and no antimicrobial peptides were used as a control. After incubating for 15 minutes, samples were taken and serially diluted and then coated on a YPD solid plate. Incubate at a constant temperature of 35°C and count the number of colonies.
[0034] Table 2 Bactericidal efficiency of anti-Candida albicans polypeptide
[0035]
[0036] The bactericidal effects of the anti-Candida albicans polypeptides are shown in Table 2. It can be seen from Table 2 that 83.78% of Candida albicans can be killed after being treated with 100 μg / mL polypeptide for 15 minutes, and 98.98% of Candida albicans can be killed after being treated with 200 μg / mL polypeptide for 15 minutes. It can ...
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Abstract
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