Process for producing feed antimicrobial peptides by bacillus subtilis and application thereof

A Bacillus subtilis and antimicrobial peptide technology, applied in the field of biological fermentation, can solve the problems of low product titer, long production cycle, complex process, etc., and achieve the effects of broad application prospects, short fermentation cycle and low cost of raw materials

Inactive Publication Date: 2017-02-22
上海邦成生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the process of producing antibacterial peptides by Bacillus subtilis still has problems such as high cost, complicated process, long production cycle, and low product titer.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] In this embodiment, the carbon source, nitrogen source and culture conditions supplemented in the fermenter in the process of producing antimicrobial peptides for feed are as follows:

[0034] (1) Carbon source: glucose 5%;

[0035] (2) Nitrogen source: yeast extract powder 4%;

[0036] (3) Fermentation process conditions: fermentation temperature 30°C; stirring speed 300r / min; ventilation ratio 1:1.5; tank pressure 0.03-0.08Mpa.

[0037] The technique of producing feed antimicrobial peptide in the present embodiment comprises the following steps:

[0038] Step 1: Cloning the silkworm antimicrobial peptide CAD gene into the pPIC9K plasmid to form a recombinant plasmid, then transforming the obtained recombinant plasmid into Bacillus subtilis, and then screening out a bacterial strain stably expressing the antimicrobial peptide CAD to obtain a production strain, namely Bacillus subtilis BW115, It is placed in a liquid nitrogen tank and stored for later use;

[0039] S...

Embodiment 2

[0044] In the present embodiment, the supplementary carbon source and culture conditions in the fermenter in the process of producing antimicrobial peptides for feed are as follows:

[0045] (1) Carbon source: 3% glucose, 3% sucrose;

[0046] (2) Fermentation process conditions: fermentation temperature 32°C; stirring speed 200r / min; ventilation ratio 1:1.2; tank pressure 0.03-0.05Mpa.

[0047] The technique of producing feed antimicrobial peptide in the present embodiment comprises the following steps:

[0048] Step 1: Cloning the silkworm antimicrobial peptide CAD gene into the pPIC9K plasmid to form a recombinant plasmid, then transforming the obtained recombinant plasmid into Bacillus subtilis, and then screening out a bacterial strain stably expressing the antimicrobial peptide CAD to obtain a production strain, namely Bacillus subtilis BW115, It is placed in a liquid nitrogen tank and stored for later use;

[0049] Step 2: Sterilize the basic fermentation medium at 121...

Embodiment 3

[0054] In the present embodiment, the supplementary carbon source and culture conditions in the fermenter in the process of producing antimicrobial peptides for feed are as follows:

[0055] (1) Carbon source: lactose 5%, sucrose 0.5%;

[0056] (2) Fermentation process conditions: fermentation temperature 37°C; stirring speed 280r / min; ventilation ratio 1:1.1; tank pressure 0.03-0.08Mpa.

[0057] The technique of producing feed antimicrobial peptide in the present embodiment comprises the following steps:

[0058] Step 1: Cloning the silkworm antimicrobial peptide CAD gene into the pPIC9K plasmid to form a recombinant plasmid, then transforming the obtained recombinant plasmid into Bacillus subtilis, and then screening out a bacterial strain stably expressing the antimicrobial peptide CAD to obtain a production strain, namely Bacillus subtilis BW115, It is placed in a liquid nitrogen tank and stored for later use;

[0059] Step 2: Sterilize the basic fermentation medium at 121...

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Abstract

The invention discloses a process for producing feed antimicrobial peptides by bacillus subtilis and an application of the process for producing the feed antimicrobial peptides by the bacillus subtilis, and bacillus subtilis BW115 of which the collection number is CCTCC (China Center For Type Culture Collection) M 2016402. The process of the invention uses BW115 to express silkworm antimicrobial peptides, researches fed-batch high-density fermentation process, carries out multi-stage amplification culture to pilot production, and then comprehensively optimizes fermentation production process conditions, so that the biological potency of an obtained fermentation liquid reaches more than 10,000U /ml, the water contents of antimicrobial peptide products obtained through spray drying are smaller than 5%, and the biological potency of the antimicrobial peptide products reaches more than 80,000U/g. The antimicrobial peptides disclosed by the invention are applied to feed adding, so that production performances of bred animals can be obviously improved; compared with the existing similar product, the process for producing the feed antimicrobial peptides by the bacillus subtilis has the characteristics of high product biological potency, low cost of raw materials, short fermentation period, simple process and suitability for large-scale industrial production.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, and in particular relates to a process for producing antimicrobial peptides for feed using Bacillus subtilis and its application. Background technique [0002] Lactose, glucose, sucrose, maltose, corn flour, corn starch, dry corn steep liquor, corn steep liquor, etc. are commonly used carbon sources for the fermentation industry. Bovine peptone, fish peptone, soybean peptone, tryptone, casein peptone, peptone, hydrolyzed milk protein, yeast extract, yeast extract powder, yeast powder, beef extract, beef extract powder, soybean meal powder, soybean cake powder, etc. are commonly used Nitrogen source for fermentation industry. Carbon sources, nitrogen sources, inorganic salts, trace elements, and growth factors are the components that make up the basic fermentation production medium. By supplementing the carbon source and nitrogen source in the basic fermentation production medium...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12P21/00A23K20/147C12R1/125
CPCC07K14/43586
Inventor 高潮陆克文郑云峰柯永中
Owner 上海邦成生物工程有限公司
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