Production method of antibacterial peptide

A production method and antimicrobial peptide technology, which is applied in the production field of antimicrobial peptides, can solve problems such as inability to separate and purify, low product titer, and low yield, and achieve the effects of good practicability, increased titer, and increased yield

Pending Publication Date: 2020-05-08
宁夏乙征生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the killing effect of antimicrobial peptides on bacteria, prokaryotic expression systems cannot be used to directly express biologically active antimicrobial peptides, and if they are expressed in the form of fusion proteins, the expression prod

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0018] Example 1

[0019] A method for producing antimicrobial peptides, the specific production steps are as follows:

[0020] S1. Put the bacteria expressed in the form of fusion protein during the preparation of antimicrobial peptides into a 300L seed tank for seed culture. After the bacteria reaches the conditions for subsequent culture inoculation, they are transferred to a 3m3 fermentor for fermentation, and the bacteria are subsequently cultured and inoculated The conditions are 6 hours of seed age, pH 5.6, OD 6.0; the end conditions of the fermentation culture of the strain are: 18 hours cycle, pH 5.9, OD 12.7; then transfer the fermentation mash to the transfer tank, and wait for the mash When the temperature in the transfer tank drops to 20-25℃, keep the mash at this temperature and let it stand for 16 hours in the transfer tank. After the standing is over, add diatomaceous earth to the transfer tank, and the amount of diatomite added is according to Add 4% of the volume...

Example Embodiment

[0022] Example 2

[0023] A method for producing antibacterial peptides, the specific production steps are as follows:

[0024] S1. Put the bacteria expressed in the form of fusion protein during the preparation of antimicrobial peptides into a 3000L seed tank for seed culture. After the bacteria reaches the conditions for subsequent culture inoculation, they are transferred to a 20m3 fermentor for fermentation, and the bacteria are subsequently cultured and inoculated The conditions are 6 hours of seed age, pH 5.6, OD 6.0; the end conditions of the fermentation culture of the bacteria are: 18 hours cycle, pH 5.8, OD 12.1; then transfer the fermentation mash to the transfer tank, and wait for the mash When the temperature in the transfer tank drops to 20-25℃, keep the mash at this temperature and let stand for 20 hours in the transfer tank. After the standing is finished, add diatomaceous earth to the transfer tank. The amount of diatomaceous earth added is according to Add 3% of ...

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PUM

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Abstract

The invention discloses a production method of an antibacterial peptide. For a fermentation mash produced by fermentation of bacteria expressed in a form of fusion protein, an antibacterial peptide inthe fermentation mash can be effectively separated from other culture medium residues and thalli by a low-temperature standing and plate-frame filtering combined method, so that the antibacterial body can be purified, the titer of an obtained product is larger than or equal to 100,000 U/g, the yield is larger than or equal to 85%, and the cost is smaller than or equal to 35 Yuan/kg. Compared withantibacterial peptides produced by other methods on the market, the antibacterial peptide prepared by the production method has a greatly improved titer, an improved yield and a substantially reducedcost, breaks through the process bottleneck of mass production of the antibacterial peptide for feed by the method, and has better practicability.

Description

technical field [0001] The invention relates to the field of production of antibacterial peptides, in particular to a production method of antibacterial peptides. Background technique [0002] Antimicrobial peptides are a class of basic small molecule peptides with antibacterial activity, with a molecular weight of about 2000-7000. Most of these active small molecule peptides have broad-spectrum antibacterial characteristics, and are the best substitutes for banned antibiotics in the feed farming industry. [0003] Due to the early ban on antibiotics in feed in Europe and the United States, the research, development and application of antimicrobial peptides for feed are far ahead of China. Due to the protection of intellectual property rights and other issues, there have been almost no exchanges on antimicrobial peptide technology, whether international or domestic. At present, the antimicrobial peptides used as feed additives in the domestic market are all imported products...

Claims

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Application Information

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IPC IPC(8): C12P21/00C07K1/34
CPCC12P21/00C07K1/34
Inventor 孙维国陈龙
Owner 宁夏乙征生物工程有限公司
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