Method for preparing antibacterial peptide by inducing fly maggots through salmonella

A Salmonella and antimicrobial peptide technology, applied in the field of biological preparation, can solve problems such as technical difficulties, complex structures, and difficulties in the separation and purification of antimicrobial peptides, and achieve good curative effect and simple and effective preparation methods

Pending Publication Date: 2021-10-22
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the low content of natural antimicrobial peptides in nature or organisms, it brings certain difficulties to the separation and purification of antimicrobial peptides
Most antimicrobial peptides have complex structures, which not only make it difficult to identify their structures, but also lead to technical difficulties and high costs in the chemical synthesis of these substances.

Method used

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  • Method for preparing antibacterial peptide by inducing fly maggots through salmonella
  • Method for preparing antibacterial peptide by inducing fly maggots through salmonella

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Salmonella one kind of antimicrobial peptides induced maggot prepared using a method comprising the steps of:

[0037] Salmonella bacteria at 12000r / min centrifugal 10min, supernatant was discarded, normal saline, 12000r / min and then centrifuged oscillation mixing 10min, repeated twice, and finally with the bacterial pellet oscillation mixing saline, resuspended to obtain salmonella.

[0038] Salmonella taken resuspended fly larvae feeding the fly larvae placed in an environment of 25 deg.] C were fed, cultured parasites maggot obtained after 24h immune defense;

[0039] Surviving larvae maggot using tap water six times, rinsed 4 times with distilled water, water with filter paper parasite surface dry, and then with 75% ethanol for disinfection of the parasite, parasite surface after drying was placed mortar in accordance with the parasite extract by mass volume 1mg: 3mL ratio of added extract (with 0.05mol / L, pH = 5.0 ammonium acetate buffer, 0.35μg / mL of PMSF and ...

Embodiment 2

[0045] A method of preparing antimicrobial peptides using Salmonella induces fly maggot, including the following steps:

[0046] The Salmonella fluid was centrifuged at 12000 r / min for 15 min, and the saline was discarded, and the saline was added. After oscillating mixing, then centrifuged for 10 min, and then repeated 3 times. Finally, the bacteria precipitate and the physiological saline were mixed, and resuspended salmonella.

[0047] Take a resuspended Salmonella feeding fly maggot, place the fly maggot in an environment of 25 ° C for feeding, and cultured after 24 hours of cultivating fly maggot of immunofemics;

[0048] The survived fly maggate was rinsed 5 times, and then rinsed 4 times with distilled water, tank the water surface of the worm body with filter paper, and then disinfect the worms with 75% alcohol. In a mortar, an extract solution was added to the proportion of the insect mass and the extraction liquid volume of 1 mg: 4 ml (a acetate buffer of acetate buffe...

Embodiment 3

[0054] A method of preparing antimicrobial peptides using Salmonella induces fly maggot, including the following steps:

[0055] The Salmonella fluid was centrifuged at 12000 R / min for 12min, abandoned with saline, and then centrifuged at 12000 r / min after mixing. salmonella.

[0056] Take a resuspended Salmonella feeding fly maggot, place the fly maggot in a 24 ° C environment for feeding, 24 hours after cultivating fly maggots produced by immunization;

[0057] The survived flies were rinsed 5 times with tap water, then rinsed with distilled water twice, using the filter paper to take the water surface of the worm, and then disinfected with 75% alcohol, etc. In a mortar, an extract solution was added to the proportion of the extract of 1 mg of extracting liquid (0.05 mol / L, pH = 5.0 acetate buffer, 0.35 μg / ml of PMSF and 0.2% mercaptoethanol. Basedly, sufficiently ground extraction, then centrifuge at 12000R / min high-speed freezing centrifuge, take the supernatant to d...

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Abstract

The invention provides a method for preparing antibacterial peptide by inducing fly maggots through salmonella, and relates to the technical field of biological preparation. A method for preparing antibacterial peptide inducing fly maggots through salmonella comprises the following steps: feeding the fly maggots with resuspended salmonella, culturing to obtain live fly maggots capable of generating immune defense, cleaning and disinfecting, extracting with an extracting solution, centrifuging, taking supernate, carrying out boiling water bath, cooling and centrifuging, concentrating and storing a crude extract, and sequentially eluting and collecting through a gel column G75 and a gel column G25 to obtain the purified maggot antibacterial peptide. According to the invention, salmonella is used for inducing fly maggots and stimulating generation of high-expression antibacterial peptide, the crude extracted antibacterial peptide is separated and purified by a gel chromatography column, the fly maggot antibacterial peptide component with high antibacterial activity is obtained, and the preparation method is simple and easy to operate.

Description

Technical field [0001] The present invention relates to the field of biological preparation technology, in particular, to a Salmonella induced maggot prepared using the method of antimicrobial peptides, antimicrobial peptides maggot prepared by the above method, and the antimicrobial peptides maggot as therapeutic agents in chicken breeding pullorum applications industry. Background technique [0002] In recent years, as people overuse of antibiotics, making the emergence of drug resistance in many pathogenic bacteria, increasing the difficulty of treatment of some diseases, the emergence of super bacteria antibiotics can not kill. Antimicrobial peptides not only have a significant role in the anti-bacterial, anti-viral, and difficult leading to drug resistant strains, making it ideal for a new generation of drugs antimicrobial peptides. Antimicrobial peptides (Antimicrobial peptides, AMP) also known as peptide antibiotics, natural or antibiotics, is a class of small peptide mole...

Claims

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Application Information

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IPC IPC(8): C12P21/00C07K1/16C07K1/14A61K38/17A61P31/04
CPCC12P21/00C07K1/16C07K1/14A61K38/1767A61P31/04Y02A50/30
Inventor 申红高盛杰王俊刚刘彩霞王政力权希闫佳许
Owner SHIHEZI UNIVERSITY
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