A kind of Enterococcus faecium producing bacteriocin and its application

An Enterococcus faecium, bacteriocin-producing technology, applied in the directions of bacteria, applications, microorganism-based methods, etc., to achieve the effects of good thermal stability, good tolerance, and good bacteriostatic effect

Active Publication Date: 2019-05-17
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, recent studies have found that the characteristic structure of the "lanthionine ring" in the Nisin molecule can inhibit probiotics in the body, and people have begun to doubt and worry about this

Method used

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  • A kind of Enterococcus faecium producing bacteriocin and its application
  • A kind of Enterococcus faecium producing bacteriocin and its application
  • A kind of Enterococcus faecium producing bacteriocin and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Isolation and purification of lactic acid bacteria:

[0036] Using the conventional plate dilution method, weigh 1g of soybean paste samples in 10mL sterilized physiological saline and shake them fully to make a 10-fold dilution. 100 μL, spread evenly on MRS solid medium, and culture at 37°C for 24h. Pick a single colony of suspected lactic acid bacteria in MRS liquid medium and culture it at 37°C for 24h. After purification by plate streak separation, Gram staining and catalase tests were performed. Select cocci that are positive in Gram staining and negative in catalase test as tentatively designated as spherical lactic acid bacteria for puncture preservation.

[0037] The bacterium was named Enterococcus faecium R1, and it was preserved on January 20, 2016 in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures. Address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, preservation unit : In...

Embodiment 2

[0038] Embodiment 2: the screening of producing bacteriocin lactic acid bacteria:

[0039] (1) Screening of bacteriostatic lactic acid bacteria

[0040] Bacteriostatic lactic acid bacteria were screened by the Oxford cup agar diffusion method.

[0041] (1) Preparation of bacterial fermentation broth: Inoculate lactic acid bacteria strains in MRS liquid medium and culture at 37°C for 24 hours. Then transfer 2% of the inoculum into MRS liquid medium for activation of the first-generation bacteria, and culture at 37°C for 24 hours. Lactic acid bacteria are activated to three generations in the same way. The culture solution was centrifuged at 10000r / m for 10min, and the fermentation supernatant was taken for later use, and stored at 4°C.

[0042] (2) Preparation and treatment of indicator bacteria suspension: Inoculate Staphylococcus aureus into LB liquid medium and culture at 37°C for 36h-48h. Then transfer 2% of the inoculum to LB liquid medium for activation of the first-g...

Embodiment 3

[0057] Example 3: Identification of bacteriocin-producing strains

[0058] (1) Physiological and biochemical identification of lactic acid bacteria

[0059] Physiological and biochemical tests were carried out on the screened bacteriocin-producing strain R1, and its species was initially identified. According to the description of Enterococcus faecium in "Berger's Bacterial Identification Manual (Eighth Edition)" and "Common Bacterial System Identification Manual", the strain R1 was preliminarily determined to be Enterococcus faecium. The specific results are shown in Table 3.

[0060] Table 3 Physiological and biochemical identification results of Enterococcus faecium R1

[0061]

[0062] (2) 16S rDNA sequence homology analysis and identification

[0063] ⑴Extraction of total DNA and detection of purity

[0064] Genomic DNA of the tested strains was extracted by the optimized CTAB method. The specific operation steps are as follows:

[0065] ①Take 1.5 mL of the bacte...

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Abstract

The present invention relates to a bacteriocin-producing Enterococcus faecium strain. The preservation name of the strain is: Enterococcus faecium (Enterococcus faecium) R1. The preservation unit is: General Microbiology Center of China Microbiological Culture Collection Management Committee. The preservation date is January 2016. 20th, deposit number: CGMCC No.12085. The source of the strain is the traditional fermented soy bean paste from farmers. In the MRS agar medium, it was a round or oval white smooth opaque colony smaller than 1 mm; in the MRS liquid medium, it grew uniformly turbid; under the electron microscope, the bacteria were spherical, Arranged in pairs or chains, Gram staining was positive. The bacterial strain of the invention is sensitive to antibiotics, does not contain virulence genes, and belongs to safe bacterial strains. The bacteriocin produced by it is enterococcin P, which is stable under acidic conditions, has good thermal stability, and has good antibacterial effect on common intestinal pathogenic bacteria.

Description

technical field [0001] The invention belongs to the field of food biotechnology, and in particular relates to a bacteriocin-producing Enterococcus faecium and an application thereof. Background technique [0002] In recent years, food safety problems have frequently erupted, and the main reason is the growth and reproduction of pathogenic bacteria and spoilage microorganisms. Food-borne pathogenic microorganisms can cause human diseases. WHO estimates that there are tens of thousands of food-borne diseases every year in the world, endangering people's health. In order to inhibit the growth and reproduction of pathogenic microorganisms, the addition of chemical preservatives has become one of the ways to solve the problem. However, the abuse of chemical preservatives makes them remain in a large amount in feed and food, posing a serious threat to poultry, livestock and human health. At this time, biological preservatives came into being. Nisin is a natural food preservative...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C07K14/315A23L3/3526C12R1/01
CPCA23L3/3526A23V2002/00C07K14/315C12N1/02C12N1/205C12R2001/01A23V2200/10A23V2250/206A23V2250/546
Inventor 武俊瑞乌日娜王晓蕊岳喜庆陶冬冰
Owner SHENYANG AGRI UNIV
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