Tegillarca granosa hemoglobin alpha helix antibacterial peptide and application thereof
A hemoglobin, alpha helix technology, applied in the directions of hemoglobin/myoglobin, application, protein-containing food ingredients, etc., can solve the problems of unstable alpha helix structure, loss of bacteriostatic effect, and easy damage.
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Embodiment 1
[0025] Embodiment 1: Optimization of ultrasonic conditions
[0026] The diluted bacterial solution was mixed with the peptide 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 and other antibacterial parameters of the antimicrobial peptide TGH2 against Escherichia coli were determined under different conditions. According to the needs of ultrasonic sound perforation mechanism, the ultrasonic power should be controlled at 5.0W / cm as much as possible 2 In the following, the α-helical structure of the antimicrobial peptide TGH2 may remain stable.
Embodiment 2
[0027] Embodiment 2: Screening of antimicrobial peptides
[0028] First log on to the NCBI (National Center for Biotechnology Information) website to find the cockles hemoglobin sequence, and then use the Antibacterial Peptide Prediction Online Server AntiBP to predict the possible antibacterial regions of the hemoglobin sequence, and conduct an investigation on the charge, hydrophobicity and reliability of the antibacterial sequence. analyze. Finally, the amino acid sequence AEFLREKLGDKCTDR was selected and named as TGH2. The hemoglobin sequence of cockles consists of 152 amino acids, and the TGH2 sequence is located at sequence number 119-136. Using the online server APD3 to calculate the hydrophobicity of the TGH2 sequence, the total hydrophobicity ratio is 33%. Finally, the antibacterial activity of TGH2 was verified.
Embodiment 3
[0029] Embodiment 3: Escherichia coli growth curve
[0030] Inoculate 20uL of the activated strain into 20mL nutrient broth liquid medium to reach 10 2-4 CFU / mL concentration, then at 0.5W / cm 2 After ultrasonic treatment for 1 hour, 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, there was no obvious damage to Escherichia coli after low-intensity ultrasonic treatment, and the growth trend was basically consistent with that under the culture conditions.
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