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Preparation method of antimicrobial peptides

A technology of antimicrobial peptides and bacterial strains, which is applied in the field of biological products, can solve the problems of restricting the production and popularization of antimicrobial peptides, difficulties in large-scale industrial production, and low yield, and achieve the effects of shortening the production cycle, no toxic side effects, and high output

Pending Publication Date: 2017-01-04
HENAN YANGSHAO BIOCHEM ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the prior art, antimicrobial peptides are produced by injecting Escherichia coli into tussah silkworm chrysalis to induce the production of antimicrobial peptides, and then extracting natural antimicrobial peptides from tussah silkworm chrysalis. The defect of difficulty in large-scale industrial production limits the production and application of antimicrobial peptides to a certain extent

Method used

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  • Preparation method of antimicrobial peptides
  • Preparation method of antimicrobial peptides
  • Preparation method of antimicrobial peptides

Examples

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Embodiment 1

[0049] The preparation method of the antimicrobial peptide provided in this embodiment specifically includes the following steps:

[0050](1) Strain rejuvenation, specifically:

[0051] First, inoculate the strain in the form of freeze-dried powder on the slant medium, and cultivate it at 32°C for 48 hours for rejuvenation;

[0052] The slant medium, in terms of mass percentage, has the following formula: 0.4% peptone, 0.2% beef extract, 2% glucose, 0.3% potassium dihydrogen phosphate, 1.6% agar, pH = natural.

[0053] (2) Prepare the seed solution, specifically:

[0054] Inoculate the rejuvenated strain in step (1) into the sterilized shake flask culture medium, and culture the shake flask at 32°C and 180 rpm for about 24 hours until the OD 600 =7, it can be considered that the culture of the strain is mature, that is, the seed solution has been prepared;

[0055] Described shaking flask culture medium, by mass percentage, formula is:

[0056] Peptone 0.4%, beef extract 0...

Embodiment 2

[0089] The preparation method of the antimicrobial peptide provided in this embodiment specifically includes the following steps:

[0090] (1) Strain rejuvenation, specifically:

[0091] First, inoculate the strain in the form of freeze-dried powder on the slant medium, and cultivate it at 31°C for 48 hours for rejuvenation;

[0092] The slant medium, in terms of mass percentage, has a formula of: 0.5% peptone, 0.25% beef extract, 1% glucose, 0.2% potassium dihydrogen phosphate, 1.8% agar, pH = natural.

[0093] (2) Prepare the seed solution, specifically:

[0094] Inoculate the rejuvenated strain in step (1) into the sterilized shake flask culture medium, and culture the shake flask at 31°C and 220 rpm for about 23 hours until the OD 600 =3, it can be considered that the culture of the bacteria is mature, that is, the seed solution has been prepared;

[0095] Described shaking flask culture medium, by mass percentage, formula is:

[0096] Peptone 0.5%, beef extract 0.25%,...

Embodiment 3

[0116] The preparation method of the antimicrobial peptide provided in this embodiment specifically includes the following steps:

[0117] (1) Strain rejuvenation, specifically:

[0118] First, inoculate the strain in the form of freeze-dried powder on the slant medium, and cultivate it at 31°C for 48 hours for rejuvenation;

[0119] The slant medium, in terms of mass percentage, has the following formula: 0.8% peptone, 0.3% beef extract, 3% glucose, 0.5% potassium dihydrogen phosphate, 1.6% agar, pH = natural.

[0120] (2) Prepare the seed solution, specifically:

[0121] Inoculate the rejuvenated strain in step (1) into the sterilized shake flask culture medium, and culture the shake flask at 31°C and 220 rpm for about 24 hours until the OD 600 =5, it can be considered that the culture of the bacteria is mature, that is, the seed solution has been prepared;

[0122] Described shaking flask culture medium, by mass percentage, formula is:

[0123] Peptone 0.8%, beef extrac...

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Abstract

The invention belongs to the technical field of biological products, and particularly relates to a preparation method of antimicrobial peptides. The antimicrobial peptides are prepared by using a Bacillus subtilis YS439 strain. The method comprises the following steps: rejuvenating the strain, preparing a seed solution, preparing a primary seeding tank culture fluid, fermenting the strain, pretreating the fermentation liquid, extracting the antimicrobial peptides, refining and the like. The specific fermentation strain culture medium and fermentation conditions are optimized to achieve the expected effects of high density and high yield; the average antibacterial titer of the fermentation liquid is 0.13-0.15 million U / ml and 0.159 million U / ml at the maximum; and the comprehensive recovery rate of the product is up to 80% or above. The production technique is simple and easy to implement, well shortens the production cycle of antimicrobial peptides, and lays a certain foundation for the popularization and application of antimicrobial peptides due to the advantages of lower production cost, high antimicrobial peptide yield, high purity and the like.

Description

technical field [0001] The invention belongs to the technical field of biological products, and in particular relates to a patent application for a preparation method of an antimicrobial peptide. Background technique [0002] Antimicrobial peptides are small molecular polypeptides with biological activity induced in organisms, and have broad-spectrum bactericidal effects. Natural antimicrobial peptides have been found to have A, B and D and other types. Most of these active polypeptides have good acid and alkali resistance, heat stability, and broad-spectrum antibacterial properties. Antimicrobial peptides have a strong killing effect on bacteria, especially on certain drug-resistant pathogenic bacteria. Compared with traditional antibiotics, the biggest feature of antimicrobial peptides is that their bactericidal mechanism is that their hydrophobic terminal helices are directly inserted into the bacterial cell membrane to form pores, and the bacterial contents are leaked ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C12R1/125
CPCC12P21/00C12N1/205C12R2001/125
Inventor 孙利鹏管宁焦国宝曹超峰席华强陈晓宇肖东坡曹忠洋何景阳马爱丽
Owner HENAN YANGSHAO BIOCHEM ENG
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