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Enterococcus faecium producing bacteriocin and application thereof

A technology of Enterococcus faecium and bacteriocin production, which is applied in the field of food biology to achieve good thermal stability, good tolerance, and good antibacterial effect

Active Publication Date: 2016-07-20
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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  • Enterococcus faecium producing bacteriocin and application thereof
  • Enterococcus faecium producing bacteriocin and application thereof
  • Enterococcus faecium producing bacteriocin and application thereof

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 and shake them fully in 10mL of sterilized normal saline for 10-fold dilution, and take samples at 10-5g / mL, 10-6g / mL, and 10-7g / mL 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 is named Enterococcus faecium (Enterococcus faecium) R1, and it was preserved on January 20, 2016 in the General Microbiology Center of China Committee for the Collection of Microbial Cultures. Address: No. 3, Yard No. 1, Beichen W...

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, according to the inoculation amount of 2%, they were transferred to the MRS liquid medium for the activation of the first-generation bacteria, and cultured at 37°C for 24h. 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 liq...

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) 16SrDNA 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 bacter...

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Abstract

The invention relates to an Enterococcus faecium strain producing bacteriocin. The strain was preserved as a name Enterococcus faecium R1 by China General Microbiological Culture Collection Center on January 20, 2016, with preservation number CGMCC No.12085. A source of the strain is farm traditional fermented soybean paste and is presented as a circular or elliptic white smooth nontransparent colony smaller than 1mm in an MRS agar culture medium; uniform turbid growth is presented in an MRS liquid culture medium; thalli are spherical under an electron microscope and are arranged in pairs or in a chain, and gram staining is positive. The strain provided by the invention is sensitive to antibiotics, does not contain virulence genes and is a safe strain. The bacteriocin produced by the strain is enterococcin P, is stable under acidic conditions, has a very good thermal stability and has a good bacteriostatic effect on common enteropathogenic 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 Applications(China)
IPC IPC(8): C12N1/20C12N1/02C07K14/315A23L3/3526C12R1/01
CPCA23L3/3526A23V2002/00C07K14/315C12N1/02C12N1/205C12R2001/01A23V2200/10A23V2250/206A23V2250/546
Inventor 乌日娜王晓蕊武俊瑞岳喜庆陶冬冰
Owner SHENYANG AGRI UNIV
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