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Method for separating plant antimicrobial peptide, plant antimicrobial peptide and application of plant antimicrobial peptide

A plant antibacterial peptide and antibacterial peptide technology, which is applied in the field of plant antibacterial peptides and plant antibacterial peptides, can solve the problems of unsystematic research on the applicability of antibacterial peptides, the need for further development of plant antibacterial peptides, and the reduction of antibacterial activity, so as to save cumbersome Process, reduced application, effect of improving production efficiency

Pending Publication Date: 2019-08-09
吉林省诚仁达医疗技术发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The applicability of antimicrobial peptides from different sources has not been systematically studied
Especially phytoantimicrobial peptides are unstable and easily hydrolyzed by proteolytic enzymes, thereby greatly reducing their antibacterial activity
Therefore, the research on plant antimicrobial peptides still needs to be deepened.

Method used

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  • Method for separating plant antimicrobial peptide, plant antimicrobial peptide and application of plant antimicrobial peptide
  • Method for separating plant antimicrobial peptide, plant antimicrobial peptide and application of plant antimicrobial peptide
  • Method for separating plant antimicrobial peptide, plant antimicrobial peptide and application of plant antimicrobial peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] A production method for directly extracting antimicrobial peptides from black sesame seeds by using ultrasonic waves:

[0065] 1. Weigh 100g black sesame seeds;

[0066] 2. Add 500 mL of 7% acetic acid solution 5 times the volume of black sesame to black sesame;

[0067] 3. The black sesame and acetic acid solution are sonicated in an ultrasonic generator with a power of 200W-300W and a frequency of 40kHz, and the extraction time is 20min, and the extract is filtered;

[0068] 4. Centrifuge the extract at 4°C for 5 minutes at 10000 rpm, discard the precipitate, so as to harvest the supernatant;

[0069] 5. Heat the extract in a water bath at 80°C for 10 minutes, and then cool to room temperature;

[0070] 6. Dialyze the supernatant for 48 hours and concentrate to obtain a reverse phase chromatography sample;

[0071] 7. Subjecting the reverse phase chromatography sample to reverse phase high performance liquid chromatography, wherein the reverse phase chromatography column is a rev...

Embodiment 2

[0075] A production method for directly extracting antimicrobial peptides from black sesame seeds by using ultrasonic waves:

[0076] 1. Weigh 100g black sesame seeds;

[0077] 2. Add 1000 mL of 5% acetic acid solution 10 times the volume of black sesame to black sesame;

[0078] 3. The black sesame and acetic acid solution are sonicated in an ultrasonic generator with a power of 200W-300W and a frequency of 40kHz, the extraction time is 35min, and the extract is filtered;

[0079] 4. Centrifuge the extract at 4°C for 15 min at 8000 rpm, discard the precipitate, so as to harvest the supernatant;

[0080] 5. Heat the extract in a 60°C water bath for 30 minutes, and then cool to room temperature;

[0081] 6. Dialyze the supernatant for 72 hours and concentrate to obtain a reverse phase chromatography sample;

[0082] 7. Subjecting the reverse phase chromatography sample to reverse phase high performance liquid chromatography, wherein the reverse phase chromatography column is a reverse phas...

Embodiment 3

[0086] A production method for directly extracting antimicrobial peptides from black sesame seeds by using ultrasonic waves:

[0087] 1. Weigh 100g black sesame seeds;

[0088] 2. Add 500 mL of 1% acetic acid solution 20 times the volume of black sesame in the black sesame;

[0089] 3. The black sesame and acetic acid solution are sonicated in an ultrasonic generator with a power of 200W-300W and a frequency of 40kHz, and the extraction time is 10min, and the extract is filtered;

[0090] 4. Centrifuge the extract at 12000 rpm at 4°C for 10 min, discard the precipitate, in order to harvest the supernatant;

[0091] 5. Heat the extract in a water bath at 100°C for 15 minutes, and then cool to room temperature;

[0092] 6. Dialyze the supernatant for 24 hours and concentrate to obtain a reverse phase chromatography sample;

[0093] 7. Subjecting the reverse phase chromatography sample to reverse phase high performance liquid chromatography, wherein the reverse phase chromatography column is...

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Abstract

The invention provides a method for separating a plant antimicrobial peptide, the plant antimicrobial peptide and application of the plant antimicrobial peptide. The method for separating the plant antimicrobial peptide comprises the following steps that a certain mass of black sesame is taken, and an acetic acid solution is added into the black sesame; the black sesame and the acetic acid solution are subjected to ultrasonic treatment in an ultrasonic generator and then filtered to obtain an extract; the extract is heated in a water bath and then cooled to the room temperature; the extract iscentrifuged, a precipitate is discarded, and a supernatant is collected; the supernatant is dialyzed and concentrated in sequence to obtain a reverse phase chromatographic sample; the reverse phase chromatographic sample is subjected to reverse phase high performance liquid chromatography to obtain the isolated plant antimicrobial peptide. The method for separating the plant antimicrobial peptide, the plant antimicrobial peptide and application of the plant antimicrobial peptide have the advantages that the black sesame is used as a raw material for direct extraction, the cumbersome processesof peeling and degreasing are omitted, application of organic reagents is reduced, the operation difficulty is reduced, the production efficiency is improved, the production cost is reduced, and themethod is in accordance with industrial production. The prepared black sesame antibacterial peptide has a broad-spectrum antibacterial effect and a strong antibacterial effect on candida albicans, pseudomonas aeruginosa, staphylococcus aureus, escherichia coli and haemophilus influenzae and can be widely applied in in-vitro killing products, poultry and livestock feed additives and other fields.

Description

Technical field [0001] The present invention relates to the field of biopharmaceuticals, in particular, to a method for separating plant antibacterial peptides, plant antibacterial peptides and uses of plant antibacterial peptides. Background technique [0002] Antimicrobial peptides are short peptides of a type of antimicrobial and some malignant cells produced by the organism in the defense response against pathogenic microorganisms. It is an important part of the body's innate immune system and has a wide range of inhibitory effects on bacteria, fungi, parasites, viruses, and tumor cells. Natural antimicrobial peptides are usually small-molecule cationic polypeptides composed of 12-60 amino acids. Because they are rich in basic amino acids such as lysine, arginine and histidine, antimicrobial peptides usually have 2-7 positive charges, etc. The electric point is greater than 7, showing strong cationic characteristics. [0003] Studies have found that the immune cells of animal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C07K1/14C07K1/20A01P1/00A01P3/00
CPCC07K14/415
Inventor 徐阳王紫嫣王国强董兵齐达王放
Owner 吉林省诚仁达医疗技术发展有限公司
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