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Bacillus subtilis, Ericin1a antibacterial peptide secreted by bacillus subtilis and application of Ericin1a antibacterial peptide

A Bacillus subtilis, antimicrobial peptide technology, applied in the field of bioengineering, can solve problems such as difficulty in improving meat quality and price, excessive meat antibiotics and chemical drug residues, and threats to health, etc., to improve the level of life science technology, protect The effect of food safety and no antibiotic residues

Active Publication Date: 2022-01-11
河北省畜牧兽医研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to prevent and treat bacterial infectious diseases such as pig bacterial diarrhea and chicken salmonellosis, various antibiotics and chemical drugs are used in large quantities in farms, resulting in excessive antibiotic and chemical drug residues in meat, which not only threatens people's health, but also Make it difficult to improve the quality and price of meat

Method used

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  • Bacillus subtilis, Ericin1a antibacterial peptide secreted by bacillus subtilis and application of Ericin1a antibacterial peptide
  • Bacillus subtilis, Ericin1a antibacterial peptide secreted by bacillus subtilis and application of Ericin1a antibacterial peptide
  • Bacillus subtilis, Ericin1a antibacterial peptide secreted by bacillus subtilis and application of Ericin1a antibacterial peptide

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Experimental program
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Effect test

Embodiment 1

[0034] 1.1 Isolation, screening and identification of high-efficiency antimicrobial peptide probiotic strains

[0035] For strain isolation and screening, samples were taken from the soil in a certain place in Pinggu, Beijing, and Bacillus subtilis was isolated. First weigh 1-10g of soil, pour it into a sterilized Erlenmeyer flask, add 50-200ml of sterilized double-distilled water, stir with a glass rod wiped with 70% alcohol cotton, and shake it on a vortex mixer for 1 -10 minutes to fully release the microorganisms in the soil into the solution. Because Bacillus subtilis can produce spores with strong stress resistance and can tolerate high temperature, high temperature treatment is used to kill other heat-labile soil microorganisms, thereby selectively screening Bacillus subtilis.

[0036] Put the Erlenmeyer flask into a 75-95°C water bath and cook for 12-20 minutes. This process will kill most microorganisms that are not resistant to high temperatures. After the end, the...

Embodiment 2

[0057] Bacteriostatic activity test

[0058] The growing single colonies were picked out separately, and after expanded culture, the antibacterial activity against Gram-negative pathogenic bacteria isolated from chicken intestines and feces was tested.

[0059] The detection of mutagenic strains with antibacterial effect adopts the classic inhibition zone method, and the specific method is as follows:

[0060] (1). Inoculate the Bacillus subtilis test bacteria and the control bacteria into 5mL LB liquid medium, and cultivate them at 37°C and 200r / min for 12h-16h.

[0061] (2). The Salmonella isolated from the feces of sick chickens was inoculated and streaked on LB solid medium, and cultivated overnight at 37°C.

[0062] (3). Pick a single colony of Salmonella with an inoculation needle and inoculate it into LB liquid medium, and culture it at 37°C and 200r / min for 12-24h.

[0063] (4). The stock solution of the cultured Salmonella is diluted 10 times with sterile water, and...

Embodiment 3

[0070] Biosafety experiment

[0071] To carry out the mouse safety experiment, the Bacillus subtilis GF158 expressing the Ericin1a antimicrobial peptide was added to the mouse food with different bacterial quantities, and the 4-week-old BALB / c mice of SPF grade were selected and randomly divided into 4 groups, 10 in each group. mice were fed continuously for 2 weeks and weighed. After the experiment, the mice in the control group and the experimental group were dissected, and the liver, kidney, spleen and lung were weighed respectively.

[0072] The experimental results showed that the weight ratio of each organ of the test mice with different doses added in the feed was not much different from that of the control group ( image 3 ), showing that Bacillus subtilis GF158 does not cause side effects of organ enlargement in experimental mice. In terms of overall weight gain, the overall weight gain speed of the experimental group added with 1-fold dose, 10-fold dose and 50-fold...

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Abstract

The invention discloses bacillus subtilis with the preservation number of CGMCC No.23153. The bacillus subtilis is a strain with the antibacterial activity further enhanced, wherein the strain is obtained through mutation screening, so that the bacillus subtilis can continuously secrete and express antibacterial peptide in animal intestinal tracts, has the advantages of probiotics and antibacterial activity small peptide, has no toxic or side effect and no antibiotic residue, replaces traditional antibiotics to be used, and improves the life science and technology level. Ericin1a antibacterial peptide can replace antibiotics to be used for preparing bacteriostatic agents for preventing bacterial diseases, and plays an important role in reducing abuse of antibiotics. The invention discloses application of Ericin1a antibacterial peptide in preparation of bacteriostatic agents for gram-negative bacteria. The invention discloses application of bacillus subtilis in preparation of livestock feed for preventing or treating gram-negative bacterial infectious diseases. The invention discloses livestock feed, and bacillus subtilis is added into the livestock feed.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a Bacillus subtilis, an Ericin1a antimicrobial peptide secreted by the Bacillus subtilis and an application thereof. Background technique [0002] The abuse of antibiotics is a very serious problem in the farming industry. Not only caused the residues of antibiotics in meat, eggs and milk, affecting people's health, but also in the past decade, the rapid emergence of superbugs that can tolerate multiple antibiotics has become a global concern, which has prompted people to look for antibiotics. Alternative antibacterial agents for livestock and poultry farming. Antimicrobial peptides (AMPs), produced by bacteria, insects, amphibians, and mammals, as well as chemically synthesized, are new antimicrobial agents to replace antibiotics due to their natural antibacterial properties, no residue, and low tendency to develop resistance to microorganisms strong candidate. Accordi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C07K14/32C12N15/31A23K10/18A23K50/30A23K50/75C12R1/125
CPCC07K14/32A23K10/18A23K50/30A23K50/75
Inventor 魏忠华杨波李杰峰李茜刘亚娟陈赛娟王珏贾国新王圆圆张绍兵高玉梅
Owner 河北省畜牧兽医研究所
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