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Pediococcus pentosaceus antagonistic to escherichia coli O157: H7 and application thereof

A technology of Pediococcus pentosaceus and Escherichia coli, applied in the field of microorganisms, can solve problems such as threats to human health, and achieve the effect of safe, reliable and high safety of food sources

Active Publication Date: 2021-10-22
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the overuse and irregular use of antibiotics in recent years, many resistant microorganisms and super-resistant bacteria have emerged, which will pose a serious threat to human health.

Method used

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  • Pediococcus pentosaceus antagonistic to escherichia coli O157: H7 and application thereof
  • Pediococcus pentosaceus antagonistic to escherichia coli O157: H7 and application thereof
  • Pediococcus pentosaceus antagonistic to escherichia coli O157: H7 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Isolation and identification of embodiment 1 lactic acid bacteria

[0031] 1. Isolation of lactic acid bacteria

[0032] Collect rutabagas in Manzhouli City, Hulunbeier City, Inner Mongolia Autonomous Region, China as a sample. Under a sterile environment, take 0.1g of rutabaga samples and add 5ml of MRS liquid medium. After shaking and mixing, they are enriched and cultured under anaerobic conditions at 37°C for 24 hours. Aspirate 0.5ml bacterial solution for gradient dilution. Add saline to make 10 -1 to 10 -5 Dilute the gradient bacterial suspension, draw 10 -3 、10 -4 、10 -5 100 μl of the three gradient bacterial suspensions were added to the MRS agar medium, spread evenly with a spreader stick, and then cultured under anaerobic conditions at 37°C for 48h. Pick the colony with typical morphology on the plate and put it on the MRS agar medium for streaking and purification. After two purifications, pick a single colony and inoculate it into the MRS liquid medium...

Embodiment 2

[0038] Example 2 Evaluation of the antibacterial ability of Pediococcus pentosaceus IM96

[0039] 1. Preparation of lactic acid bacteria fermentation broth

[0040] A single colony of Pediococcus pentosaceus IM96 was inoculated in MRS broth medium and cultured anaerobically at 37°C for 48h. Centrifuge at 10000r×4°C for 10min to obtain the fermentation supernatant. The fermentation supernatant was filtered with a 0.22 μm filter membrane to obtain the cell-free supernatant from the fermentation of Pediococcus pentosaceus IM96, which was frozen at -80°C until use.

[0041] 2. Preparation of indicator bacteria

[0042] Inoculate single colonies of Escherichia coli O157:H7 ATCC 43895 in LB liquid medium, culture at 37°C, 200r / min for 6h, adjust the concentration to 1×10 8 CFU / mL for antibacterial experiments.

[0043] 3. Determination of antibacterial ability of Pediococcus pentosaceus IM96

[0044] Take 200 μL of the indicator bacteria solution prepared above and spread it ev...

Embodiment 3

[0045] Example 3 Evaluation of the probiotic performance of Pediococcus pentosaceus IM96

[0046] 1. Artificial gastric juice tolerance experiment of Pediococcus pentosaceus IM96

[0047] Tolerance to artificial gastric juice test: adjust the pH value of PBS (pH7.4) buffer solution to 3.0 with 0.1mol / L HCl, add pepsin (3g / L), dissolve it and filter it with a 0.22μm sterile filter membrane. Ready to match. Activate the screened strains, inoculate the prepared Pediococcus pentosaceus IM96 bacterial solution in 10mL MRS liquid medium with a volume ratio of 2% inoculum, culture at 37°C for 24 hours under anaerobic conditions, and collect by centrifugation at 4000×g for 10 minutes Bacteria were washed with PBS (pH7.4) for 3 times, and the concentration of the bacteria solution was adjusted to 10 with PBS. 9 CFU / mL, spare. Take 1 mL of PBS-resuspended lactic acid bacteria liquid and add it to 5 mL of simulated gastric juice, and at the same time add 1.5 mL of 0.5% (w / v) NaCl to m...

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Abstract

The invention discloses pediococcus pentosaceus antagonistic to escherichia coli O157: H7 and application of the pediococcus pentosaceus antagonistic to escherichia coli O157: H7. The pediococcus pentosaceus is pediococcus pentosaceus IM96 and is preserved in the Guangdong Microbial Culture Collection Center on May 17, 2021, with an accession number being GDMCC NO: 61654. Viable bacteria and fermentation supernatant of the pediococcus pentosaceus have good antibacterial activity, and especially have strong antagonistic activity on escherichia coli O157: H7 in vitro and in vivo. In addition, the pediococcus pentosaceus has good tolerance to gastrointestinal fluid, is high in safety, and can relieve and improve intestinal pathological injury, intestinal inflammation and intestinal epithelial barrier injury caused by escherichia coli O157: H7 in vivo. The pediococcus pentosaceus can be used for preparing medicines, food or food additives, animal feeds and microbial preparations with antibacterial activity in the future, and has good application prospects.

Description

Technical field: [0001] The invention belongs to the technical field of microbes, and in particular relates to a Pediococcus pentosaceae with antibacterial activity and an application thereof. Background technique: [0002] Enterohaemorrhagic Escherichia coli (EHEC), a pathogenic subgroup of Shiga toxin-producing Escherichia coli (STEC), produces Shiga toxin 1 and / or 2 (Stx1 and Stx2) and often contains an adhesion gene (eae). Escherichia coli O157:H7 is the most common pathogenic serotype of enterohaemorrhagic Escherichia coli. Enterohemorrhagic Escherichia coli can cause systemic complications such as bloody diarrhea, hemorrhagic enteritis, hemolytic uremic syndrome, and thrombotic thrombocytopenic purpura. Since enterohaemorrhagic Escherichia coli was first discovered in the United States in 1982, food contamination by Ehemorrhagic Escherichia coli has occurred in many countries in the world, resulting in many deaths. In 1996, the world's largest outbreak of Escherichia...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/689C12N15/11A23K10/18A23L33/135A61K35/74A61P31/00C12R1/01
CPCC12N1/20C12Q1/689A23L33/135A23K10/18A61K35/74A61P31/00A23V2002/00A23V2400/427A23V2200/30Y02A50/30
Inventor 吴清平李海新李滢谢新强陈惠元张菊梅丁郁王涓陈谋通薛亮张淑红杨小鹃韦献虎张友雄
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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