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Preparation method for antimicrobial peptide in coelomic fluid of sipunculus nudus

A technology of star worm body cavity and grid star worm, which is applied in the field of active ingredient extraction, can solve the problems of extracting antibacterial substances that have not been found, and achieve the effects of shortening hydrolysis time, low production cost, and improving resource utilization efficiency

Inactive Publication Date: 2019-01-11
AGRI PRODS PROCESSING RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the research on the extraction of antibacterial substances from the coelom fluid of Starworms

Method used

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  • Preparation method for antimicrobial peptide in coelomic fluid of sipunculus nudus
  • Preparation method for antimicrobial peptide in coelomic fluid of sipunculus nudus
  • Preparation method for antimicrobial peptide in coelomic fluid of sipunculus nudus

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preparation example Construction

[0026] A preparation method for antimicrobial peptides in the coelom fluid of Atragalus, comprising the following steps:

[0027] Steps for cleaning and spit sand: select fresh and vigorous square star worms, wash them with sterilized seawater; feed the square star worms in light salt water with a mass concentration of 24‰, change the water for feeding every 1 hour, and wait for the square star worms The starworm spits out the sand;

[0028] Steps for collecting body cavity fluid: Take out the square starworm that has spit out all the sediment, rinse it with sterile water, use filter paper to absorb excess water on the body surface, hold the front part of the starworm with tweezers, and make it perpendicular to the sterile centrifuge tube, place it in the sterile centrifuge tube Cut a small hole at the end of the worm body to let the body fluid flow out naturally, and collect them one by one;

[0029] Ultrasonic crushing steps: Ultrasonic crushing and cracking of the collecte...

Embodiment 1

[0037] After washing and feeding the fresh phalaenopsis, the body cavity fluid is collected, the cells are broken by ultrasonic waves, centrifuged to obtain the crude protein extract of the body cavity fluid, and then enzymatically hydrolyzed to obtain the body cavity fluid antimicrobial peptide;

[0038] Step 1: Select fresh and vigorous square star worms, wash them with sterilized seawater; feed the square star worms in light salt water with a mass concentration of 24‰, change the water every 1 hour, and wait for them to spit out all the sediment;

[0039] Step 2: Take out the star worm that has exhausted the silt, rinse it with sterile water, absorb the excess water on the body surface with filter paper, hold the front part of the star worm with tweezers, make it perpendicular to the sterile centrifuge tube, and cut a piece at the end of the worm body Small mouth, let the body fluid flow out naturally, collect one by one.

[0040] Step 3: Ultrasonic cracking of the body cav...

Embodiment 2

[0044] After washing and feeding the fresh starworms, the body cavity fluid is collected, the cells are broken by ultrasonic waves, centrifuged to obtain the crude protein extract of the body cavity fluid, and then the antimicrobial peptide of the body cavity fluid is obtained by enzymatic hydrolysis;

[0045] Step 1: Select fresh and vigorous square star worms, wash them with sterilized seawater; feed the square star worms in light salt water with a mass concentration of 24‰, change the water every 1 hour, and wait for them to spit out all the sediment;

[0046]Step 2: Take out the star worm that has exhausted the silt, rinse it with sterile water, absorb the excess water on the body surface with filter paper, hold the front part of the star worm with tweezers, make it perpendicular to the sterile centrifuge tube, and cut a piece at the end of the worm body Small mouth, let the body fluid flow out naturally, collect one by one.

[0047] Step 3: Ultrasonic cracking of the body...

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Abstract

The invention discloses a preparation method for an antimicrobial peptide in coelomic fluid of sipunculus nudus. The preparation method comprises the following steps: cleaning and purifying; collecting coelomic fluid; ultrasonically breaking; centrifugally filtering; performing enzymolysis treatment, thereby acquiring a target antimicrobial peptide in coelomic fluid of sipunculus nudus. The invention provides a mild and efficient cell disruption process by adopting an integrated process of ultrasonic technology and enzymolysis and utilizing low ultrasonic treatment splitting decomposition to acquire intracellular protein; a great convenience is supplied for hydrolysis extraction for subsequent large-scale preparation of protein; protease is adopted for performing enzymolysis treatment on the coelomic fluid of sipunculus nudus; the antimicrobial peptide with high antibacterial activity is acquired by controlling the time, temperature and enzyme addition of enzymolysis.

Description

technical field [0001] The invention relates to a technology for extracting active ingredients, in particular to a method for preparing antibacterial peptides in coelom fluid of Acarina graminis. Background technique [0002] Antimicrobial peptides refer to class 1 small molecular polypeptides that exist in organisms, can resist external microorganisms, and eliminate mutant cells in the body. Antimicrobial peptides have broad-spectrum antibacterial properties, especially for killing multidrug-resistant bacteria. It has a different mechanism of action from antibiotics, and bacteria are not easy to develop resistance to it; the problem of multi-drug-resistant bacteria caused by the abuse of antibiotics has become a global public health problem that threatens human health. To develop a natural, pollution-free, efficient and safe antibacterial Drugs are an important development direction, which is of great significance to alleviate the spread of multi-drug resistant bacteria an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/34
CPCC07K1/34C12P21/06Y02P20/10
Inventor 曹玉坡李积华代亚萍周伟韩志萍李亚会
Owner AGRI PRODS PROCESSING RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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