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Bacillus coagulans FM 603 and application thereof

A technology of Bacillus coagulans and strains, which is applied in the field of microorganisms and their cultivation and application, can solve the problem of lactic acid bacteria losing the protection function of mice, etc., and achieve the effect of promoting animal production performance and improving breeding efficiency.

Active Publication Date: 2015-12-23
沈阳丰美生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bacteriocin is an important functional substance of probiotics. Researchers at University College Cork found that bacteriocin-producing lactic acid bacteria can effectively reduce the infection of Listeria monocytogenes in mice, but the bacteria obtained in Listeria monocytogenes Lactic acid bacteria lose their protective function in mice after immunization with bacteriocin

Method used

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  • Bacillus coagulans FM 603 and application thereof
  • Bacillus coagulans FM 603 and application thereof
  • Bacillus coagulans FM 603 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: Analysis of the antibacterial activity of Bacillus coagulans FM603 bacteriocin

[0031] Streak FM603 on LB agar plate, culture overnight at 30°C, pick a single colony and inoculate in 10mL LB medium, 150rpm, culture at 30°C for 24h, inoculate 0.2% inoculum in 1000mL modified tryptone medium, culture at 150rpm, 30°C After 24 hours, centrifuge at 10,000 rpm for 15 minutes to obtain the fermentation supernatant. Streak L.innocua, L.monocytogenes, S.aureus, B.cereus, and M.luteus on LB agar plates and culture overnight at 37°C, pick a single colony and inoculate it in LB medium, culture overnight at 37°C, and pipette 10ul The culture solution was inoculated on 10mL LB soft agar (LB medium containing 0.75% agar), spread evenly on the LB agar plate, placed in a sterilized Oxford cup after cooling, and added 100μl FM603 fermentation supernatant or two-fold gradient dilution The fermentation supernatant was cultured overnight at 30°C, the inhibition zone was obse...

Embodiment 2

[0034] Embodiment 2: the preparation of FM603 bacteriocin

[0035] 1. Fermentation culture

[0036] Streak culture of Bacillus coagulans FM603 on the LB plate, pick a single colony and inoculate it in 10mL liquid LB medium for overnight culture, inoculate 10mL overnight culture seed solution in 1000mL improved tryptone medium, its composition is: mass volume percentage, Tryptone 1.5%, glucose 1.5%, yeast extract powder 0.5%, corn steep liquor dry powder 0.2%, ammonium sulfate 0.05%, sodium chloride 0.1%, sodium dihydrogen phosphate 0.05%, distilled water 1000mL, pH7.0, culture condition is 30 Cultivate at 150 rpm for 20 hours.

[0037] 2. Preparation of Bacteriocin Crude Extract

[0038] The cultured FM603 fermentation broth was centrifuged at 10000×g for 15 min to take the supernatant for later use. Equilibrate the ion column with 50mM, pH 6.5 phosphate buffer, the fermentation supernatant passes through the ion column at a flow rate of 50mL / min, discard the supernatant th...

Embodiment 3

[0041] Embodiment 3: the biological characteristic of FM603 bacteriocin

[0042] 1) Tolerance to temperature: Treat the fermentation supernatant of Example 2 at 60, 70, 80, 90, and 100°C for 10 minutes respectively, take the sample without heat treatment as a control, and L.innocua as an indicator bacterium to detect heat treatment For the effect on bacteriocin activity, the antibacterial activity retention rate is represented by the ratio of the diameter of the inhibition zone of the treatment group to the diameter of the control group. The results are shown in Table 2. The bacteriocin activity is basically Unaffected, the activity remained above 90%.

[0043] 2) Sensitivity to enzymes: get the fermentation supernatant of Example 2 and add pepsin, trypsin, pronase, lipase, amylase respectively, the final concentration of enzyme is 1mg / mL, and the pepsin treatment pH is 2.5, The pH of other enzyme treatment was 6.5, and after 2 hours of treatment at 37°C, L.innocua was used a...

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Abstract

The invention provides a new strain FM 603 of bacillus coagulans, and the preservation number of the strain is CGMCC NO.10221. The strain can generate bacteriocin antibacterial substances and achieves antibacterial activity for Gram-positive pathogenic bacteria such as Listeria monocytogenes, staphylococcus aureus, methicillin-resistant staphylococcus aureus, clostridium perfringens and clostridium difficile. The molecular weight of bacteriocin is 4276.45 Da, the partial amino acid sequence is Ala-Gly-His-Dhb-Phe-Val-Dhb-Gly-Pro, and the bacteriocin is stable under treatment of heat, acid, and pepsase or trypsin and can be degraded easily by pronase to be inactivated. The invention further provides an FM603 fermentation product which can be used as a microbiological feed additive. Animal test results show that the fermentation product can increase the laying rate of laying hens, reduce the feed-egg ratio and improve egg quality; the feed intake and daily gain of piglets are increased, and the feed conversion ratio is reduced; the daily gain and the content of lysozyme in serum of broiler chickens are increased, and the feed conversion ratio and the death rate are reduced.

Description

technical field [0001] The invention relates to microorganisms and their cultivation and application technology, in particular to a new strain of bacillus coagulans producing bacteriocin and the application of the strain in the field of feed additives. Background technique [0002] Antibiotics are secondary metabolites produced by microorganisms that have antimicrobial activity against other microorganisms. In the 1940s, scientists discovered that feeding animals low doses of antibiotics could improve animal performance and increase economic efficiency. In the following decades, antibiotics were widely used in the animal husbandry industry, added to feed as a growth promoter, or used as a therapeutic agent in the breeding field. The use of antibiotics has significantly improved the growth efficiency and feed conversion rate of animals, which has played an important role in the development of the animal husbandry industry. But the use of antibiotics has also brought many pr...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K1/16C12R1/07
Inventor 郭耀棋丁毅王丽娟曹岩峰唐日益
Owner 沈阳丰美生物技术有限公司
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