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Enterococcus durans and generated bacteriocin for inhibiting listeria monocytogenes

A technology of durable enterococcus and listeria, applied in the field of microorganisms, can solve the problems of difficult promotion and high equipment, and achieve the effect of simple method, high efficiency and low cost

Active Publication Date: 2013-09-18
NANJING UNIV OF FINANCE & ECONOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, some scholars have also carried out research on the control of Listeria using ultra-high pressure sterilization technology. However, due to the high requirements of this technology on equipment, it is difficult to promote it in industry.

Method used

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  • Enterococcus durans and generated bacteriocin for inhibiting listeria monocytogenes
  • Enterococcus durans and generated bacteriocin for inhibiting listeria monocytogenes
  • Enterococcus durans and generated bacteriocin for inhibiting listeria monocytogenes

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0040] Screening and Identification of Example 1 Durable Enterococcus (Enterococcus durans) NJ Strain

[0041] MRS medium contains: 9.0g / L peptone, 6.0g / L beef extract powder, 4.5g / L yeast extract powder, 18.0g / L glucose, 1.0mL / L Tween 80, 2.0g / L dipotassium hydrogen phosphate, 5.0g / L sodium acetate, 2.0g / L triammonium citrate, 0.2g / L magnesium sulfate, 0.06g / L manganese sulfate, the pH value is 6.5.

[0042] After 10-fold gradient dilution of traditional fermented dairy products from Inner Mongolia, spread them on the MRS medium, isolate single colonies, control the number of single colonies in each plate to be between 100-400, and culture overnight in a 37°C incubator. Then pick up the culture medium covered with single colonies in the plate, put it horizontally on the agar plate inoculated with 0.5% fresh Listeria (L.monocytogenes CMCC54004, purchased from China Institute for the Control of Pharmaceutical and Biological Products, the same below), and continue to cultivate ...

Embodiment 2

[0044] Implementation example 2 Preparation of bacteriocin produced by Enterococcus durans NJ strain

[0045] (1) Preparation of fermentation broth of Enterococcus durans NJ strain

[0046] Seed medium formula: glucose 20.0g / L, peptone 10.0g / L, beef extract powder 10.0g / L, yeast extract powder 5.0g / L, Tween-801.0mL / L, dipotassium hydrogen phosphate 2.0g / L, Sodium acetate 5.0g / L, diamine hydrogen citrate 2.0g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.05g / L, pH value 6.2, solvent is water.

[0047] Fermentation medium 1 formula: glucose 18.0g / L, peptone 9.0g / L, beef extract powder 6.0g / L, yeast extract powder 4.5g / L, Tween-801.0mL / L, dipotassium hydrogen phosphate 2.0g / L , sodium acetate 5.0g / L, triammonium citrate 2.0g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.06g / L, pH value 6.5, solvent is water.

[0048] Pre-treatment of fermentation medium 1: Rinse AMBERLITE XAD16 resin (purchased from Sigma) with pH 2, 70% by volume isopropanol aqueous solution, and then us...

Embodiment 3

[0071] Example 3 Properties of Bacteriocins Produced by Enterococcus durans NJ Strain

[0072] After the fermentation broth 1 in Example 2 was centrifuged, the obtained fermentation broth supernatant was subjected to the following property determination.

[0073] (1) pH stability: the pH value of the supernatant of Enterococcus durans NJ strain fermentation broth was adjusted to pH 2, 3, 4, 5, 6, 7, 8, 9, 10 and 11 with hydrochloric acid, and then Incubate at 37°C for 2 hours. Using the double dilution method, drop 5ul of the supernatant of the fermentation broth on the BHI solid medium inoculated with 0.5% fresh Listeria, cultivate overnight, and observe the maximum dilution that can inhibit the growth of Listeria. It was found that under various pH conditions, the maximum dilution of the supernatant of the fermentation broth that could inhibit the growth of Listeria was 8 times. The results indicated that the bacteriocin produced by the NJ strain of Enterococcus durans had...

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Abstract

The invention provides enterococcus durans, and bacteriocin which is generated from Enterococcus durans and is used for inhibiting listeria monocytogenes, and relates to the field of biology. The preservation registration number of the enterococcus durans NJ strain is CGMCCNo.7236. The bacteriocin is generated from the NJ strain and is used for inhibiting listeria monocytogenes. The preparation method of the bacteriocin comprises the following steps of: fermenting and culturing the NJ strain, obtaining a fermentation liquid, and purifying the fermentation liquid so as to obtain the bacteriocin. The bacteriocin which is generated from the NJ strain and which is used for inhibiting listeria monocytogenes has good pH and thermal stability, and has instantaneous high-efficient inhibition function only for listeria monocytogenes, so that the bacteriocin can be applied to food fresh-keeping. The bacteriocin is prepared by fermenting and purifying the NJ strain, and the method is simple, high in efficiency, low in cost and safe.

Description

technical field [0001] The invention relates to a strain of enterococcus durabilis in the field of microbes and the bacteriocin produced by it which can inhibit Listeria monocytogenes. Background technique [0002] Bacteriocins are a class of proteins or polypeptides with bacteriostatic activity produced by ribosomes, which widely exist in Gram-positive and Gram-negative bacteria, and their inhibition of target bacteria is characterized by low concentration and high efficiency. Especially since the first commercial application of nisin, people have paid more attention to the research of bacteriocins. Due to the biological safety status of lactic acid bacteria bacteriocins, the application of bacteriocins as biological preservatives meets consumers' requirements for safety, health and naturalness as more and more attention is paid to the potential safety hazards of chemical preservatives. [0003] Listeriosis is an intestinal infectious disease with a high fatality rate caus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C07K14/315C12P21/02C07K1/36C07K1/34C07K1/18C12R1/46
Inventor 都立辉鞠兴荣袁建何荣王立峰宋伟刘琴
Owner NANJING UNIV OF FINANCE & ECONOMICS
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