Broad-spectrum antibacterial Bacillus amyloliquefaciens strain and application thereof

A technology of Bacillus amyloliquefaciens and antibacterial solution, applied in the field of microorganisms, can solve problems such as food safety and environmental pollution, and achieve the effects of stable bacteriostatic performance, strong tolerance and simple culture conditions

Active Publication Date: 2019-03-08
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention provides a kind of bacillus amyloliquefaciens JFL21 which can effectively antagonize variou...

Method used

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  • Broad-spectrum antibacterial Bacillus amyloliquefaciens strain and application thereof
  • Broad-spectrum antibacterial Bacillus amyloliquefaciens strain and application thereof
  • Broad-spectrum antibacterial Bacillus amyloliquefaciens strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Separation and purification of bacterial strains

[0038] Purchase aquatic products and food (freshly dissected hairtail intestines, brewed tea and fermented soybean meal) from Wushan Garden Vegetable Market, Wushan Road, Tianhe District, Guangzhou, add appropriate saline for grinding, and treat in a 70°C water bath for 30 minutes, then take 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 The diluted solution was spread on an LB medium plate and incubated at a constant temperature of 37°C for 24h. A single colony was picked and streaked on an LB plate for isolation and repeated purification. Carry out Gram staining and microscopic examination of the purified colonies, pick out rod-shaped Gram-positive strains, and number them, then use glycerol with a final concentration of 20% as a protective agent, and store them in a -80°C ultra-low temperature refrigerator for later use .

Embodiment 2

[0039] Embodiment 2: Screening of antagonistic bacteria

[0040] (1) Preparation of Bacillus fermentation supernatant: use an inoculation loop to take a ring of liquid bacteria stored at -80°C and streak it on an LB solid plate, and culture overnight in a 37°C incubator. Pick a single colony of activated Bacillus and inoculate it in a 250mL Erlenmeyer flask filled with 50mL of LB medium, and culture it at 37°C and 200r / min for 24h to obtain an activated seed solution. The seed solution was transferred to a 250mL Erlenmeyer flask containing 100mL Landy fermentation medium according to the inoculum amount of 1%, and fermented at 30°C and 200rpm for 48h. The fermentation broth was centrifuged at 4°C and 12,000 rpm for 15 minutes to remove bacteria, and then filtered through a 0.22 μm water filter membrane to obtain a sterile fermentation supernatant, which was used for the detection of antibacterial activity.

[0041] (2) Preparation of indicator bacteria plate:

[0042] Retran...

Embodiment 3

[0045] Embodiment 3: strain identification

[0046] (1) Morphological identification: The strain JFL21 obtained by screening was isolated on LB plates by streaking, and the colony characteristics were observed after culturing at 37°C for 48 hours. The results were as follows: figure 1 As shown, the colony is milky white, round, with irregular edges, wrinkled surface, inset in the middle, opaque, and relatively dry; after being cultured in LB liquid medium for 24 hours, the single colony is purple after Gram staining, which is leather. Lambert-positive bacteria, rod-shaped, the results are shown in figure 2 .

[0047] (2) Physiological and biochemical characteristics: Classification and identification of bacterial species described in "Berger's Bacteria Identification Manual" (8th edition) and "Common Bacterial System Identification Manual" (edited by Dong Xiuzhu, Cai Miaoying, etc., Beijing: Science Press, 2001.2) The physiological and biochemical characteristics of the str...

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Abstract

The invention discloses a broad-spectrum antibacterial Bacillus amyloliquefaciens strain and application thereof, and belongs to the technical field of microbes. The strain is named as Bacillus amyloliquefaciens JFL21, and is collected in the Guangdong Microbial Culture Collection Center on November 5, 2018, with the collection number of GDMCC NO: 60473. The Bacillus amyloliquefaciens JFL21 is cultured under a simple culture condition, is easy to preserve and strong in stress resistance, is easily produced industrially, and has good development and application prospects; a fermented supernatant thereof has broad-spectrum antibacterial activity for 19 common foodborne pathogens and aquatic pathogens, has extremely stable bacteriostatic performance for Listeria monocytogenes and Aeromonas hydrophila, can be further developed as a probiotic, a biological preservative or an antibacterial medicine, and is applied to food preservation or prevention of aquaculture diseases, so that use of antibiotics is reduced to a certain extent, and an application prospect is broad.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a strain of Bacillus amyloliquefaciens JFL21 with broad-spectrum antibacterial ability, and the application of the strain in preventing and treating food-borne pathogens and aquatic pathogens. Background technique [0002] Pathogenic microorganisms are widely distributed in nature, and their hazards have seriously affected the healthy development of various industries. First of all, pathogenic microorganisms are the main factors that cause food spoilage and food-borne diseases. In the process of food processing, storage and transportation, food spoilage often occurs due to the contamination of pathogenic microorganisms. According to statistics, more than 25% of food is wasted every year due to pathogenic microorganism contamination worldwide, causing economic losses of tens of billions of dollars each year (Papadopoulou et al., 2014; Yuan Jianguo, 2011). In addition...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23L3/3571A61K35/742A61P31/04C12R1/07
CPCA23L3/3571A23V2002/00A61K2035/115A61P31/04C12N1/205C12R2001/07A23V2200/10Y02A50/30
Inventor 林俊芳林龙镇郭丽琼郑倩望魏韬
Owner SOUTH CHINA AGRI UNIV
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