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Method for producing antibacterial peptide by using brevibacillus laterosporu

A technology of Bacillus lateralis and Bacillus brevis, applied in the field of biological fermentation, can solve the problems of few reports on antimicrobial peptides, poor heat resistance, and narrow antibacterial spectrum, achieve broad-spectrum antibacterial activity, good heat resistance, and reduce production cost effect

Active Publication Date: 2012-07-04
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many microorganisms can produce antimicrobial peptides. The current research mainly focuses on lactic acid bacteria. However, most of the antimicrobial peptides derived from lactic acid bacteria have problems such as narrow antibacterial spectrum and poor heat resistance. more urgent
The antibacterial peptides produced by Bacillus and Paenibacillus generally have broad antibacterial spectrum and high stability, which have begun to attract the attention of researchers at home and abroad, but there are not many reports on the antibacterial peptides produced by Bacillus and Paenibacillus

Method used

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  • Method for producing antibacterial peptide by using brevibacillus laterosporu
  • Method for producing antibacterial peptide by using brevibacillus laterosporu
  • Method for producing antibacterial peptide by using brevibacillus laterosporu

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Effect test

Embodiment 1

[0034] The preparation of embodiment 1 antimicrobial peptide

[0035] 1. Preparation of seed solution

[0036] Inoculate Brevibacillus laterosporus AS1.2738 into the sterilized seed culture medium and culture it at 30°C for 16 hours to obtain a seed solution with a bacterial content of 10 8 cfu / ml.

[0037] The preparation of the seed medium is as follows: yeast extract 5g, peptone 10g, sodium chloride 5g, distilled water 1000ml, pH 7.3;

[0038] 2. Preparation of antimicrobial peptides by fermentation

[0039] The seed solution obtained in step 1) was inoculated into the fermentation medium according to the volume ratio of the inoculation amount 2% to carry out fermentation culture, culture conditions: 32 ° C, shaker speed 180r / min, fermentation culture 18h,

[0040] The fermentation medium contains 3.5% of carbon source, 3% of nitrogen source and 0.2% of inorganic salt, all in mass-volume ratio.

Embodiment 2

[0041] The preparation of embodiment 2 antimicrobial peptides

[0042] 1. Preparation of seed solution

[0043] Inoculate Brevibacillus laterosporus AS1.2739 into the sterilized seed culture medium and culture it at 37°C for 12 hours to obtain a seed solution with a bacterial content of 10 7 cfu / ml.

[0044] The preparation of the seed medium is as follows: beef extract 10g, peptone 20g, sodium chloride 7g, distilled water 1000ml, pH 7.2;

[0045]2. Preparation of antimicrobial peptides by fermentation

[0046] The seed solution obtained in step 1) was inoculated into the fermentation medium according to the volume ratio of the inoculation amount of 2% to carry out fermentation culture, culture conditions: 35 ° C, shaking table speed 280r / min, fermentation culture 16h,

[0047] The fermentation medium contains 0.5% of carbon source, 0.5% of nitrogen source and 0.005% of inorganic salt, all in mass-volume ratio.

Embodiment 3

[0049] The determination of antimicrobial peptides adopts the internationally recognized "protease sensitive method", and the fermented liquids obtained in Example 1 and Example 2 are respectively processed as follows: 1. 10,000rpm centrifuged for 15min to remove the bacterium to obtain the supernatant, and then pass through a 0.22 μm filter membrane 2. Adjust the pH of the supernatant to 6.5 to get rid of the effect of organic acids; 3. Take an equal amount of sample solution and mix with 10mg / mL protease (pepsin, protease E, trypsin, protease K) The solution was mixed in 1:1 volume, reacted in a 37°C constant temperature water bath for 2h, and boiled in a 100°C boiling water bath for 5min to kill the enzyme after the reaction; 4. Take the samples before and after protease treatment respectively, and use the agar diffusion method to measure the inhibition zone diameter and compare. The results are shown in Table 1, and the results show that the bacteriostatic substances obtai...

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Abstract

The invention provides a method for producing antibacterial peptide. The method comprises the following steps of: inoculating brevibacillus laterosporu serving as a strain into a fermentation culture medium consisting of a carbon source, a nitrogen source and inorganic salt; and cultivating in a shaking table at the temperature of 30 to 40 DEG C for 12 to 24 hours to obtain the antibacterial peptide with valence of 1,000 to 1,500 AU / ml. The antibacterial peptide has wide antibacterial spectrum, high safety and high stability. The method for producing the antibacterial peptide and the product have the advantages of low cost, simple preparation process, high yield, wide antibacterial spectrum, high sterilizing efficacy and the like and can be widely applied in the fields of preservation and corrosion prevention of food, vegetables, fruits, feed additives and the like.

Description

technical field [0001] The invention relates to the field of biological fermentation, in particular to a process for preparing antimicrobial peptides by fermentation of Bacillus lateralis. Background technique [0002] Antimicrobial peptides (AMP) are biologically produced polypeptides with antibacterial activity, generally with a molecular weight of less than 5kD and an amino acid number of less than 100. Compared with antibiotics, antimicrobial peptides are safe, harmless to normal cells of humans and animals, and belong to a class of biological preservatives that have no toxic side effects, no residue, and no bacterial resistance. It has a broad antibacterial spectrum, not only can inhibit bacteria, but also has a certain inhibitory effect on fungi, viruses and cancer cells. In addition, it also has the characteristics of high stability, good heat resistance and good solubility. These advantages make it have broad application prospects in the food industry and feed indu...

Claims

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Application Information

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IPC IPC(8): A23J3/20A23L3/3526A23K1/17A23K20/195
Inventor 贾英民李兴峰王志新王广贤孙继良田丰伟陈卫
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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