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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

Active Publication Date: 2007-09-26
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The yield of natural scorpion venom is extremely low, and there are many types of active peptides with very similar structures. It is very difficult to separate and purify single components, which seriously restricts the further development and research of this potential active substance.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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PUM

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Abstract

The invention discloses a structure, function and making method of karsch analgesic antibiotic active peptide, whose amino acid sequence is NGYLLDKYTGCKVWCVINNESCNSECKIRGG-YYGYCYFWK LACFCQGARNSELWNYNTNKCNGKL, wherein the active peptide or derivant or analog can be synthesized through gene engineering technique, which solves the difficulty to separate and purify peptide; the active peptide is mixed with multiple carriers as medical acceptable agent, which is fit for clinical.

Description

Technical field: [0001] The present invention relates to the field of biotechnology. Specifically, it uses biotechnology to screen, separate and purify from scorpion, scorpion tail or scorpion venom, discover and obtain an analgesic and antibacterial active peptide and its preparation method. The active peptide has analgesic and antibacterial biological activities. Background technique: [0002] Scorpions are the oldest surviving arthropods in the world, and their morphology has changed little during the evolutionary process of more than 400 million years. The main means for scorpions to capture prey and defend against natural enemies is toxins secreted by the tail glands, which act on ion channels on the surface of living cell membranes to paralyze or kill enemies. In the long evolutionary process, scorpions rely on scorpion toxins with multiple biological activities. Survival, and scorpion toxin has also become a natural active peptide library rich in a variety of functional pe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/36C12N15/12C12N15/63A61K38/17A61P29/00C07H21/04
CPCY02A50/30
Inventor 张景海邵建华刘岩峰张嵘吴春福
Owner SHENYANG PHARMA UNIVERSITY
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