Scorpion analgesic antibacterial active peptide and preparation method thereof
An antibacterial activity and pH value technology, applied in the field of analgesic antibacterial active peptides and their preparation methods, can solve the problems of difficulty in single-component separation and purification, restricting the development and research of active substances, low yield of natural scorpion venom, and the like, and achieving a simple preparation method. Effect
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Embodiment 1
[0024] Example 1: Purification and preparation of scorpion analgesic and antibacterial active peptides using scorpion venom as raw material
[0025] After dissolving 2.0 g of scorpion venom in 20 ml of distilled water, the supernatant was separated at 15,000 rpm for 15 minutes, and the pH of the supernatant was adjusted to 2 with 0.01M hydrochloric acid or phosphoric acid solution. 15,000 rpm was separated for 15 minutes, and the supernatant was taken with sodium phosphate The pH of the solution was adjusted to 13, 15,000 revolutions per centimeter for 15 minutes. Take the supernatant and adjust the pH 4-9 with aqueous hydrochloric acid or phosphoric acid, and load the sample on a gel filtration chromatography column (1.6cm×66cm) that has been pre-equilibrated with 20mM sodium phosphate buffer (pH under physiological conditions, containing 150mM NaCl) , Such as: Sephacryl S-100HR or Superose 12 prep grade or Superdex 30 prep grade or Superdex75pre-p grade or Sephadex G-50 or Sepha...
Embodiment 2
[0026] Example 2: Purification and preparation of scorpion analgesic and antibacterial active peptides using scorpion tail as raw material
[0027]After 100 g of scorpion tail was homogenized with 0.5 liter of distilled water, the supernatant was collected at 4500 revolutions / centre for 20 minutes. After the precipitation was homogenized with 0.5 liter of distilled water, the supernatant was collected at 4500 revolutions / centre for 20 minutes, and the precipitation was added 0.5 After homogenizing liters of distilled water, 4,500 revolutions / centre separated for 20 minutes, the supernatant was collected, the above three extracts were combined, and the pH was adjusted to 2, with 0.01M hydrochloric acid or phosphoric acid solution, 5000 revolutions / detached 15 minutes, and the supernatant was taken Adjust the pH to 13,5000 rpm with sodium phosphate solution for 15 minutes. Other operations are the same as in Example 1.
Embodiment 3
[0028] Example 3: Purification and preparation of scorpion analgesic and antibacterial active peptides using whole scorpion as raw material
[0029] After homogenizing 500 g of scorpion with 2.5 liters of distilled water, 4,500 revolutions / centre separated for 20 minutes, and the supernatant was collected. After the precipitation was homogenized with 1.5 liters of distilled water, the supernatant was collected at 4500 revolutions / separated for 20 minutes, and the supernatant was collected and 1.0 was added for precipitation. After homogenizing liters of distilled water, 4,500 revolutions / centre separated for 20 minutes, the supernatant was collected, the above three extracts were combined, and the pH was adjusted to 2, with 0.01M hydrochloric acid or phosphoric acid solution, 5000 revolutions / detached 15 minutes, and the supernatant was taken Adjust the pH to 13,5000 rpm with sodium phosphate solution for 15 minutes. Other operations are the same as in Example 1.
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