Bacteriocin produced by Bacillus coagulans and application thereof

A technology of Bacillus coagulans and bacteriocin, applied in the field of microorganisms, can solve problems such as poor stability, and achieve good thermal stability, good antibacterial performance, and good probiotic effect

Active Publication Date: 2019-07-19
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that nisin bacteriocin, which can be used as a green biological preservative in the prior art, only has an inhibitory effect on some Gram-positive bacteria and has poor stability, the first object of the present invention is to provide a kind of bacteriocin, and the second object Provide a broad-spectrum antibacterial agent, the third purpose is to provide the biological application of the bacteriocin, including its application in the preparation of biological preservatives and in the prevention and treatment of livestock and poultry diseases or aquatic diseases

Method used

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  • Bacteriocin produced by Bacillus coagulans and application thereof
  • Bacteriocin produced by Bacillus coagulans and application thereof
  • Bacteriocin produced by Bacillus coagulans and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Isolation and Identification of Bacillus coagulans CGMCC9951

[0026] 1. Isolation and purification of Bacillus: Collect 30 copies of cecum and feces of healthy piglets in ecological farms under aseptic conditions, weigh 1g of the samples and add them to 9mL sterile saline, shake and mix well, and put them in a water bath at 80°C Remove non-spore bacteria in a water bath for 10 minutes, inoculate in the improved LB medium according to the inoculum amount of 1%, incubate at 37°C for 20 hours, and spread on the improved calcium carbonate identification plate; select a single colony with a calcium-dissolving circle and grow well , transferred to the nutrient agar slant with an inoculation loop for pure culture, repeated subculture for 3 times, and stored in a 4°C refrigerator for later use.

[0027] 2. Observation of bacterial colony morphology: activate the slant bacterial species preserved in step 1 for 2-3 times, take a clean glass slide and carry out Gram sta...

Embodiment 2

[0033] Example 2 Stress resistance verification of Bacillus coagulans CGMCC NO.9951

[0034] 1. Tolerance test of simulated gastric acid: Inoculate Bacillus coagulans with the preservation number CGMCC NO.9951 in LB medium, culture at 37°C for 36 hours, and centrifuge the bacterial solution at 5000 rpm for 10 minutes to collect the bacterial cells. The same amount of bacteria was added to 10 mL of sterile PBS buffer with pH 1, 2, 3, 4, 5 as the test group; the bacteria were added to 10 mL of sterile PBS buffer with pH 7.0 as the control group. The above-mentioned treated samples were cultured statically at 37° C. for 2 hours, and counted on plates. The survival rate was the ratio of the number of live bacteria in the experimental group to the control group multiplied by 100%. The results showed that the survival rate was 91±3.5% at pH 4 after being exposed to the extreme condition of pH 1-2 for 2 hours. It can be seen that Bacillus coagulans has strong acid resistance and ca...

Embodiment 3

[0037] Determination of embodiment 3 bacillus coagulans optimal antibacterial effect culture time

[0038]Inoculate Bacillus coagulans CGMCC NO.9951 into the optimized LB medium, culture at 230rpm, 37°C for 16h, start at 16h of culture, take samples at intervals of 8h, continue sampling until 56h of culture, and take samples 6 times in total. The cell-free fermentation supernatant was obtained by centrifugation at 4000rpm, and the diameter of the inhibition zone was measured by using the Oxford cup antibacterial test method. Each group of experiments was repeated three times, and the average value of the diameter of the inhibition zone was taken. The results show that the antibacterial activity of the cell-free supernatant of Bacillus coagulans CGMCC9951 increases continuously with the growth of culture time, and its antibacterial activity reaches the maximum at 48 hours. The results are shown in Figure 6 .

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Abstract

The invention provides a bacteriocin produced by Bacillus coagulans and an application thereof, which belong to the field of microorganisms, the bacteriocin is extracted from of Bacillus coagulans with a preservation number of CGMCC NO. 9951, and can tolerate simulated gastric acid and simulate bile salt environment; has significant probioticity and can effectively inhibit growth and reproductionof Escherichia coli, Pastevula mulfocida, salmonella typhi, Salmonella, Shigella dysenteriae, Vibrio alginolyticus, Aeromonas hydrophila, and vibrio parahaemolyticus; and the bacteriocin has good stability and good antibacterial effect. The bacteriocin and bacteriocin-producing Bacillus coagulans can be used in the fields of prevention and control of poultry and aquatic diseases.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a bacteriocin produced by Bacillus coagulans and an application thereof. Background technique [0002] Because Bacillus can transform vegetative cells into spores with good stress resistance, it can enhance its resistance to adverse environments such as high temperature, acid and alkali, and this tolerance increases its potential as a probiotic. Since Bacillus coagulans not only has the characteristics of Bacillus coagulans, but also has the advantages of lactic acid bacteria, it is of great significance to obtain a new type of Bacillus coagulans with good probiotic properties. [0003] Bacteriocin is a kind of polypeptide or protein with antibacterial activity encoded by genes. It has the advantages of high efficiency, non-toxicity, high temperature resistance, no residue, and no drug resistance. It is used as a green biological preservative in the preservation of agric...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/32C07K1/30A61K38/16A61P31/04A23L3/3463
CPCA23L3/3463A61K38/00A61P31/04C07K14/32Y02A40/70
Inventor 吴影周子吕古绍彬李市场田晶晶李长福
Owner HENAN UNIV OF SCI & TECH
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