A kind of mud cockles hemoglobin alpha helix antimicrobial peptide and its application
A technology of hemoglobin and antimicrobial peptides, applied in hemoglobin/myoglobin, application, protein-containing food ingredients, etc., can solve problems such as loss of antibacterial effect, easy to be destroyed, unstable α helix structure, etc.
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Embodiment 1
[0025] Example 1: Optimization of ultrasonic conditions
[0026] The diluted bacterial solution and peptide were mixed and placed in an ultrasonic water bath at a temperature of 37°C. Using ultrasonic frequency, ultrasonic power and ultrasonic time as parameters, the antibacterial activity of antimicrobial peptide TGH2 against Escherichia coli and other antibacterial parameters were determined under different conditions. According to the needs of the ultrasonic sonoporation mechanism, the ultrasonic power should be controlled at 5.0W / cm as far as possible. 2 In the following, the α-helical structure of the antimicrobial peptide TGH2 may remain stable.
Embodiment 2
[0027] Example 2: Screening of antimicrobial peptides
[0028] First, log on to the website of NCBI (National Center for Biotechnology Information) to find the hemoglobin sequence of the slug, and then use the antibacterial peptide prediction online server AntiBP to predict the possible antibacterial regions of the hemoglobin sequence, and conduct the charge, hydrophobicity and reliability of the antibacterial sequence. analyze. The amino acid sequence AEFLREKLGDKCTDR was finally screened and named as TGH2. The hemoglobin sequence of the slug is composed of 152 amino acids, and the TGH2 sequence is located in the sequence number 119-136. Using the online server APD3 to calculate the hydrophobicity of the TGH2 sequence, the total hydrophobicity ratio was 33%. Finally, the antibacterial activity of TGH2 was verified.
Embodiment 3
[0029] Example 3: E. coli growth curve
[0030] Inoculate 20uL of the activated strain into 20mL of nutrient broth liquid medium to 10 2-4 CFU / mL concentration, then at 0.5W / cm 2 Under ultrasonic treatment for 1h, take 20ul (0, 1, 2, 3, 4, 5, 6, 7, 8 hours) samples for plate counting and observe the growth curve. like figure 1 As shown, the E. coli treated with low-intensity sonication did not have obvious damage, and the growth trend was basically the same as that under the culture conditions.
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