Method for producing antimicrobial peptide through fermentation of brevibacillus laterosporu

A Brevibacillus and antibacterial peptide technology, applied in microorganism-based methods, biochemical equipment and methods, fermentation and other directions, can solve the problems of large influence of pH value of fermentation broth, unsuitable for industrial production, complex production process, etc., and achieve antibacterial Wide spectrum, easy to promote, good safety effect

Active Publication Date: 2014-08-27
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Wu Zhaoliang et al [Xu Hao, Wu Zhaoliang, Yin Hao, Yan Jin. Fermentation, adsorption, separation and coupling production process of nisin. Polymer Materials Science and Engineering, 2010,26(10):155-158] reported that the fermentative production of Nisin The cation exchange resin D113 was added as an adsorbent to carry out adsorption-coupled fermentation. When the fermentation end point was reached, the titer of Nisin increased from 3350IU / mL to 5189IU / mL, which was an increase of 54.9%. Activation treatment, complex operation
[Pongtharangku T, Demirci A.Online recovery of nisin during fermentation and its effect on nisin production in biofilm reactor.Applied Microbiology and Biotechnology, 2007,74(3):555-562] using microbial fermentation and membrane separation and adsorption separation Nisin is produced by the coupling method, which increases the titer of Nisin by nearly 4 times, and the effect is remarkable, but the production process is complicated, the cost is high, and the pH value of the fermentation broth is greatly affected, so it is not suitable for industrial production

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  • Method for producing antimicrobial peptide through fermentation of brevibacillus laterosporu
  • Method for producing antimicrobial peptide through fermentation of brevibacillus laterosporu
  • Method for producing antimicrobial peptide through fermentation of brevibacillus laterosporu

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Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1 Utilizes the technology of Brevibacillus lateralsporosa fermentation to produce antimicrobial peptide

[0030] The specific process is as follows:

[0031] 1. Seed cultivation

[0032] Put the glycerol tube containing B.laterosporu AS1.2827 into the sterilized NB culture medium at 32°C for 24 hours; then insert it into the NB medium at 5v / v% inoculum size and culture at 32°C for 8 hours to obtain the bacterial concentration for 10 7 cfu / mL of the secondary seed solution.

[0033] 2. 5L fermenter culture

[0034] Put the secondary seed liquid obtained in step 1 into a 5L fermenter equipped with 3L fermentation medium at a 5v / v% inoculation amount, at 32°C, 300r / min, and under the conditions of ventilation 1:0.5v / v / min Cultivate for 15h.

[0035] Among them, fermentation medium (w / v): glycerin 2.0%, soybean meal powder 2.0%, CaCl 2 0.005%, Tween-200.5%, adjusted to pH 7.2, prepared with water.

[0036] 3. Add activated carbon adsorption separation coup...

Embodiment 2

[0040] Embodiment 2 Utilizes the technique of Brevibacillus lateralsporosa fermentation to produce antimicrobial peptide

[0041] The specific process is as follows:

[0042] 1. Seed cultivation

[0043] Put the slant of preserved B.Laterosporu AS1.2738 into the sterilized NB medium, 34°C, culture time 24h, then insert 3v / v% inoculation amount into the NB seed medium, culture at 34°C for 12h, and obtain the bacterial concentration for 10 8 cfu / mL of the secondary seed solution.

[0044] 2. Fermentation tank culture

[0045] The secondary seed liquid obtained in step 1 was inserted into a 5L fermenter equipped with 4L fermentation medium according to the inoculum amount of 8v / v% to carry out fermentation culture.

[0046] Fermentation medium (w / v): 1.5% soluble starch, 1.5% yeast powder, CaCl 2 0.15%, ZnCl 2 0.008%, MnCl 2 0.05%, Tween-202.0%, pH7.0, prepared with water.

[0047] Fermentation conditions: temperature 34°C, ventilation rate 1:0.5v / v / min, dissolved oxygen ...

Embodiment 3

[0051] The impact of embodiment 3 activated carbon addition time on viable count and antimicrobial peptide output

[0052] Using Brevibacillus lateralosporu (B.laterosporu) AS1.864 as the fermentation strain, first put the glycerol tube of the preserved strain into the sterilized NB medium, culture it at 32°C for 24 hours, and insert it into NB according to the inoculation amount of 3v / v%. Culture medium, cultivated at 32°C for 12h, and obtained a bacterial concentration of 10 8 cfu / mL of the secondary seed solution.

[0053] A 5L fermenter was used, and the inoculation amount of 5v / v% was inserted into the system equipped with 3L fermentation medium, and the fermentation was carried out at 32°C, the ventilation rate was 1:0.5v / v / min, and the rotation speed was 300r / min. At the 12th, 16th, 20th, and 24th hours of fermentation, add 2% powdered activated carbon and 2v / v% concentrated material medium, and continue to ferment for another 24 hours; with no activated carbon as a co...

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Abstract

The invention provides a method for producing antimicrobial peptide through fermentation of brevibacillus laterosporu. In fermentation of the brevibacillus laterosporu, when the yield of the antimicrobial peptide is no longer increased, active carbon is directly added into the fermentation liquor; as the active carbon adsorbs the fermentation product, the inhibiting effect of the product is relieved, and the density of the thallus and the yield of the antimicrobial peptide are greatly improved. According to the method, the active carbon is taken as an adsorption stripping coupling agent, has low cost, is environmentally friendly, and can be directly added in the fermentation process without being activated in advance; the antimicrobial peptide can be adsorbed without regulating the pH value of the fermentation liquor; and the method is simple and easy to promote, and the active carbon can be regenerated through desorption after fermentation, thus being recycled repeatedly, and lowering the production cost.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for producing antimicrobial peptides by fermentation of Brevibacillus sporogenes. Background technique [0002] With the improvement of people's living standards and the increasing emphasis on food safety, consumers demand to reduce the use of chemical preservatives and antibiotic residues. Therefore, the development of safe and efficient biological preservatives has become an important development direction of the food industry, among which antimicrobial peptides ( Antimicrobial peptide (AMP) has a potential application value in replacing chemical preservatives and antibiotics due to its chemical nature of protein, no drug residue, no immunogenicity, and no toxicity in humans or animals. [0003] Antimicrobial peptides are small molecular polypeptides widely present in organisms. The first isolated antimicrobial peptides come from the silkworm chrysalis of the silk...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C12R1/01
Inventor 贾英民王志新李兴峰宁亚维杨瑾
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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