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Lactobacillus plantarum SWUN5815 for high-yield bacteriocin and application of lactobacillus plantarum SWUN5815

A technology of Lactobacillus plantarum and Lactobacillus plantarum, applied in the field of microorganisms, can solve the problems such as narrow bacteriostatic spectrum, lactic acid bacteria cannot adapt to the geographical environment of the Qinghai-Tibet Plateau, and cannot be applied to yaks, etc.

Inactive Publication Date: 2017-05-31
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the prior art, most lactic acid bacteria produce bacteriocins with a narrow antibacterial spectrum, and can only inhibit Gram-positive bacteria or Gram-negative bacteria alone; due to species specificity, most lactic acid bacteria cannot adapt to the severe geographical environment of the Qinghai-Tibet Plateau. not suitable for yaks

Method used

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  • Lactobacillus plantarum SWUN5815 for high-yield bacteriocin and application of lactobacillus plantarum SWUN5815

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Effect test

Embodiment 1

[0029] The isolation and identification of embodiment 1 bacteria

[0030] 1. Isolation of bacteria

[0031] Weigh 2g of fresh yak feces sample and 20ml of sterile water, shake and mix evenly, and dilute them in 10-fold series. Take different dilutions respectively under aseptic conditions, and pour them in MRS (containing 0.5% calcium carbonate) medium in a plate. plate, cultured at 37°C for 48h. Pick individual colonies with calcium-dissolving circles and different shapes for Gram staining, select Gram-positive bacteria, inoculate them in MRS liquid medium, and cultivate them at 37°C. Number the lactic acid bacteria obtained after screening and put them on MRS solid medium Draw a line for purification and save for later use;

[0032] 33 strains of lactic acid bacteria were screened from 90 strains of lactic acid bacteria by the Oxford cup method, which had obvious inhibitory effects on the indicator strains. After excluding the influence of acid, there were still 9 strains ...

Embodiment 2

[0042] The antibacterial activity of embodiment 2 thalline

[0043] 1. Preparation of Lactobacillus plantarum MRS fermentation supernatant:

[0044] Inoculate the preservation solution in the cryopreservation tube into 5mL MRS liquid medium, shake and mix well, incubate at a constant temperature of 37°C for 24 hours, continuously activate three generations with 2% inoculum size, and take the third generation fermentation liquid and centrifuge at 4000×g After 10 minutes, take the supernatant, filter the supernatant through a 0.22 μm micro-membrane, store in a -20°C refrigerator for later use, and prepare the Lactobacillus plantarum MRS fermentation supernatant.

[0045] 2. Exclusion of acid inhibition

[0046] The inhibitory effect of Lactobacillus plantarum MRS fermentation supernatant on indicator bacteria may be due to the effect of acidic products secreted by lactic acid bacteria such as acetic acid and lactic acid. In order to remove the interference of acidic substances...

Embodiment 3

[0053] Embodiment 3 antibacterial spectrum is measured

[0054] Table 1 Antibacterial spectrum of bacteriocin produced by SWUN5815

[0055]

[0056] Note: Diameter of antibacterial zone (mm): +++, 19-24; ++, 15-18; +, 10-15; -, no antibacterial effect.

[0057] Common yak-derived diarrhea-causing enteric pathogens, Salmonella enteritidis, Salmonella typhimurium, Salmonella dublin, and Escherichia coli Shigagenes, were selected as indicator bacteria, and the cell-free fermentation supernatant was used to conduct the Oxford cup antibacterial test.

[0058] The results of the antibacterial spectrum of SWUN5815 bacteriocin are shown in Table 1. It can be seen from the table that the bacteriocin produced by SWUN5815 has broad-spectrum antibacterial activity, and its pathogenic indicator bacteria were all isolated from yaks. In addition to having a good inhibitory effect on the indicator bacteria used in this study, it also has a good inhibitory effect on some common pathogenic...

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Abstract

The invention discloses a lactobacillus plantarum SWUN5815 for high-yield bacteriocin and an application of the lactobacillus plantarum SWUN5815. The strain is preserved at the China Center for Type Culture Collection (CCCTCC) on August 2, 2016 and the preservation number is CCTCC NO: M2016419; and the strain is separated from yak dung. The bacteria generated by the strain can be degraded by protease; the strain has a very high inhibition effect on salmonella and escherichia coli and has good acid resistance and relatively high cholate tolerance; the survival rate after 3h in artificial gastric juice of which the pH is 3.0 is 93.36+ / -4.06%; the strain can slowly grow in 1.0% cholate; and the growth efficiency reaches 15.26+ / -1.19% of that of the strain which is cultivated under the condition free of the cholate. The bacteriocin is capable of effectively inhibiting growth of salmonella, escherichia coli and staphylococcus aureus. Therefore, the lactobacillus plantarum SWUN5815 for the high-yield bacteriocin has the potential of being applied to yak intestinal tracts as a microecological preparation.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a plantarum lactobacillus SWUN5815 for high-yield bacteriocin and its application. Background technique [0002] Lactic acid bacteria are the dominant microbial flora in the intestines of humans and animals. Probiotic lactic acid bacteria can regulate the micro-ecological balance of humans and animals, inhibit and kill pathogenic bacteria, and enhance the body's immunity. [0003] Lactobacillus plantarum is a kind of Lactobacillus found in pickles, yogurt and the gastrointestinal tract of animals, which can widely utilize plant-based carbon sources, tolerate bile salts and have low blood sugar levels. Some Lactobacillus plantarum can produce bacteriocinoids, which can inhibit intestinal pathogenic bacteria such as coliforms, Salmonella, and Shigella. The bacteriocins produced by Lactobacillus plantarum are composed of natural amino acids and are not modified after t...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00C12R1/25
CPCC12P21/00C12N1/205C12R2001/01
Inventor 汤承岳华周宴阳
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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