Preparation method of snakehead epidermal mucus antibacterial peptide

An antibacterial peptide and mucus technology is applied in the field of preparation of the mucus antibacterial peptide of the snakehead skin, which can solve the problems of high cost of the antibacterial peptide, ineffective utilization of the mucus, waste of biological resources, etc., and achieves excellent antibacterial activity, promotes sustainable development, and is good for The effect of suppressing the effect

Pending Publication Date: 2021-05-14
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chemical synthesis is currently a relatively common method in the production of antimicrobial peptides, but the high cost of artificially synthesized antimicrobial peptides makes it difficult to use them in batches in low-cost production such as living and breeding
Commonly known as snakehead, snakehead snakehead is a rare freshwater fish with high economic value and nutritional and medical value. With the development of snakehead aquaculture, a large amount of mucus produced during the processing of snakehead snakehead has not been effective...

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  • Preparation method of snakehead epidermal mucus antibacterial peptide
  • Preparation method of snakehead epidermal mucus antibacterial peptide
  • Preparation method of snakehead epidermal mucus antibacterial peptide

Examples

Experimental program
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Effect test

Embodiment 1

[0035] (1) Pre-treatment of snakehead: Injure the back of fresh snakehead, gently scrape the back with a plastic scraper, and collect the mucus every 2 hours for later use;

[0036] (2) Heat treatment: the collected snakehead skin mucus was immediately placed in hot water and heated for 5 minutes;

[0037] (3) Centrifugation: Cool the mucus heated with hot water in step (2) to below 5°C, perform high-speed centrifugation on the heat-treated mucus at a speed of 8000r / min, and centrifuge for 20 minutes, and collect the supernatant;

[0038](4) Freeze-dried storage: put the supernatant collected in step (3) into a -40°C refrigerator for pre-freezing for 6 hours, and then use a fully automatic vacuum freeze dryer to dry and make powder to obtain the lyophilized powder of snakehead skin mucus, namely for the crude extract;

[0039] (5) Use the Sephadex G-100 gel column to purify the freeze-dried powder in step (4), dissolve the freeze-dried powder in distilled water, the column si...

Embodiment 2

[0043] (1) Pre-treatment of snakehead: Injure the back of fresh snakehead, gently scrape the back with a plastic scraper, and collect the mucus every 2 hours for later use;

[0044] (2) Heat treatment: immediately place the collected snakehead skin mucus in hot water and heat for 10 minutes;

[0045] (3) Centrifugation: Cool the mucus heated with hot water in step (2) to below 5°C, perform high-speed centrifugation on the heat-treated mucus at a speed of 6000r / min, centrifuge for 40min, and collect the supernatant;

[0046] (4) Freeze-dried storage: put the supernatant collected in step (3) into a -40°C refrigerator for 6 hours, and then use a fully automatic vacuum freeze dryer to dry and make powder, and then obtain the lyophilized powder of snakehead skin mucus. is the crude extract;

[0047] (5) Use the Sephadex G-100 gel column to purify the freeze-dried powder in step (4), dissolve the freeze-dried powder in distilled water, the column size is 1.2cm×70cm, and the sample...

Embodiment 3

[0058] (1) Pre-treatment of snakehead: Injure the back of fresh snakehead, gently scrape the back with a plastic scraper, and collect the mucus every 2 hours for later use;

[0059] (2) Heat treatment: the collected snakehead skin mucus was immediately placed in hot water and heated for 5 minutes;

[0060] (3) Centrifugation: Cool the mucus heated with hot water in step (2) to below 5°C, perform high-speed centrifugation on the heat-treated mucus at a speed of 7000r / min, centrifuge for 30min, and collect the supernatant;

[0061] (4) Freeze-dried storage: put the supernatant collected in step (3) into a -40°C refrigerator for pre-freezing for 6 hours, and then use a fully automatic vacuum freeze dryer to dry and make powder to obtain the lyophilized powder of snakehead skin mucus, namely for the crude extract;

[0062] (5) Use the Sephadex G-100 gel column to purify the freeze-dried powder in step (4), dissolve the freeze-dried powder in distilled water, the column size is 1....

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Abstract

The invention belongs to the technical field of biological extraction, and relates to a preparation method of a snakehead epidermal mucus antibacterial peptide. The preparation method comprises the following steps: collecting mucus of the back epidermis of snakeheads, heating in a water bath, cooling, centrifuging and collecting supernate; after pre-freezing, freeze-drying the supernate to obtain freeze-dried powder of the snakehead epidermal mucus; purifying the freeze-dried powder by adopting a Sephadex G-100 gel column, then collecting a product, detecting a light absorption value at 280 nm, collecting a component, and freeze-drying to obtain a freeze-dried component; and further purifying by adopting a cation exchange column, collecting a product, detecting a light absorption value at 280 nm, finally collecting a component, dialyzing, desalting and freeze-drying to obtain the snakehead epidermal mucus antibacterial peptide. The method is simple in process and relatively low in cost, has a good inhibition effect on gram-positive bacteria and gram-negative bacteria, has broad-spectrum antibacterial activity, can be used for preparing a fish preservative or a feed additive, and is beneficial to realizing high-value utilization of snakehead processing byproducts.

Description

technical field [0001] The invention belongs to the technical field of biological extraction, and in particular relates to a method for preparing an antibacterial peptide from snakehead epidermis mucus. Background technique [0002] Antimicrobial peptides (AMPs) are a class of polypeptides with antibacterial activity widely distributed in mammals, amphibians, insects or other invertebrates, also known as host defense peptides, and are important components of the innate immune system part. AMPs typically range in length from 10 to 50 amino acids, have a net positive charge of 2 to 8, and exhibit broad activity against a variety of microorganisms, including Gram-positive and Gram-negative bacteria, fungi, viruses, and parasites, and are highly toxic Little or no toxicity. Antimicrobial peptides have the advantages of thermostability, broad-spectrum antibacterial, and resistance to drug resistance. Studies have found that they have the potential to replace traditional antibio...

Claims

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Application Information

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IPC IPC(8): C07K2/00C07K1/14C07K1/16C07K1/18A23K20/147A23K20/195A61K38/02A61P31/04
CPCA61K38/00A23K20/147A23K20/195A61P31/04C07K2/00
Inventor 高瑞昌伍晓云石彤储倩李欣袁丽
Owner JIANGSU UNIV
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