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Method for extracting antibacterial peptide from fermentation liquid

A fermentation broth and antimicrobial peptide technology, which is applied in the field of antimicrobial peptide extraction, can solve the problems of pathogenic bacteria drug resistance and threats to health, and achieve the effects of avoiding secondary retention effects, enhancing secretion, and increasing the speed of circulation

Inactive Publication Date: 2018-08-17
金华市艾力生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] At present, all conventional antibiotics have corresponding drug-resistant pathogenic strains, and the problem of drug resistance of pathogenic bacteria has become an increasingly serious threat to people's health

Method used

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  • Method for extracting antibacterial peptide from fermentation liquid

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Effect test

Embodiment 1

[0023] The method for extracting antimicrobial peptides from fermented liquid comprises the following steps:

[0024] 1) Bacillus subtilis was cultured with nutrient broth medium at 140r / min and 35°C for 24h; Saccharomyces cerevisiae was cultured with nutrient broth medium at 120r / min and 36°C for 24h; Aspergillus oryzae Cultivate with Chapei’s medium, culture at 160r / min, 28°C for 48h; use sterile normal saline to adjust the number of bacteria to 1.0×10 8cfu / mL; 2) With 100 parts of soybean meal as the main ingredient, 12 parts of wheat bran powder and 10 parts of corn flour as auxiliary materials, 0.8 parts of honey and 1 part of sucrose were added, and distilled water was added until the water content was 45%. 0.3 part of fermentation synergist is added after fermentation to obtain the fermentation medium; 3) Take Bacillus subtilis seed liquid, Saccharomyces cerevisiae seed liquid and Aspergillus oryzae seed liquid according to the weight ratio of 1:0.3:0.3, and mix the bac...

Embodiment 2

[0026] The method for extracting antimicrobial peptides from fermented liquid comprises the following steps:

[0027] 1) Preparation of seed solution: Bacillus subtilis was cultured with nutrient broth medium at 160r / min and 37°C for 48 hours; Saccharomyces cerevisiae was cultured with nutrient broth medium at 150r / min and 38°C 36h; Aspergillus oryzae was cultured using Chapei’s medium at 180r / min, 30°C for 72h; the number of bacteria was adjusted to 1.2×10 with sterile normal saline respectively. 8 cfu / mL; Under different culture conditions, expand the culture of Bacillus subtilis, Saccharomyces cerevisiae and Aspergillus oryzae, and use the sufficient nitrogen source, carbon source and trace elements in the nutrient medium to make each strain rapidly grow The vigorous stage prepares for further mixed fermentation;

[0028] 2) Preparation of fermentation medium: 120 parts of soybean meal as the main ingredient, 15 parts of wheat bran powder and 15 parts of corn flour as auxi...

Embodiment 3

[0035] The method for extracting antimicrobial peptides from fermentation broth comprises: preparing seed liquid, preparing fermentation medium, fermenting, ultrafiltration, and separating, specifically including the following steps:

[0036] Preparation of seed solution: Bacillus subtilis was cultured with nutrient broth medium, cultured at 150r / min, 36°C for 36h; Saccharomyces cerevisiae was cultured with nutrient broth medium, cultured at 150r / min, 37°C for 30h; Aspergillus oryzae was cultured using Chapei’s medium at 170r / min, 28°C for 48h; the number of bacteria was adjusted to 1.2×10 with sterile normal saline. 8 cfu / mL; Under different culture conditions, expand the culture of Bacillus subtilis, Saccharomyces cerevisiae and Aspergillus oryzae, and use the sufficient nitrogen source, carbon source and trace elements in the nutrient medium to make each strain rapidly grow The vigorous stage prepares for further mixed fermentation;

[0037] Preparation of fermentation med...

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Abstract

The invention discloses a method for extracting antibacterial peptide from fermentation liquid. The method comprises the following steps of preparation of seed liquid, preparation of a fermentation culture medium, fermentation, ultra-filtration and separation. The method comprises the following specific steps: firstly, culturing bacillus subtilis, saccharomyces cerevisiae and aspergillus oryzae seed liquid respectively; then, inoculating the seed liquid into a fermentation culture medium prepared from soybean meal powder, bran powder and corn flour for fermenting; performing purification and ultrafiltration on the fermentation liquid to obtain four segments of polypeptide solutions of different molecular weights; performing gel permeation chromatography separation and liquid chromatographyseparation on the polypeptide solutions in sequence to obtain the antibacterial peptide. The method has the beneficial effects that the antibacterial peptide is prepared by fermenting mixed bacteria,and four segments of antibacterial peptide of different molecular weights are obtained after separation and purification, so that waste of active polypeptide in the fermentation liquid is avoided, and the bioactivity of the polypeptide at each molecular weight segment can be researched finely and distinguishingly. Meanwhile, by adopting the method, the separation accuracy, correctness and efficiency of the antibacterial peptide are increased.

Description

technical field [0001] The invention relates to the field of extracting antimicrobial peptides, in particular to a method for extracting antimicrobial peptides from fermentation broth. technical background [0002] At present, all conventional antibiotics have corresponding drug-resistant pathogenic strains, and the problem of drug resistance of pathogenic bacteria has become an increasingly serious threat to people's health. Finding new types of antibiotics is an effective way to solve the problem of drug resistance. Antimicrobial peptides are considered to have broad application prospects in the pharmaceutical industry because of their high antibacterial activity, wide antibacterial spectrum, many types, wide range of options, and resistance mutations in target strains. At present, a variety of peptide antibiotics are undergoing preclinical feasibility studies, [0003] Microbial fermentation refers to the process of using microorganisms to convert raw materials into pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P21/00C07K1/36C07K1/34C07K1/22C07K1/16C12R1/125C12R1/865C12R1/69
CPCC12P39/00C07K1/16C07K1/22C07K1/34C07K1/36C12P21/00
Inventor 周艳初
Owner 金华市艾力生物科技有限公司
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