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Preservation method of bacillus coagulans BC99

A technology of Bacillus coagulans and glycerol bacteria, applied in the field of microorganisms, can solve the problems of difficult activation of bacteria species, cumbersome operation, and uneven quality of Bacillus coagulans products, and achieve good consistency of bacteria, simple and convenient operation, and stress resistance strong effect

Pending Publication Date: 2021-06-22
武汉微康益生菌研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the differences in strains and the impact of production processes, the quality of products on the market related to Bacillus coagulans is uneven
Using general strain preservation methods, there are disadvantages such as difficulty in later activation of strains and cumbersome operations.

Method used

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  • Preservation method of bacillus coagulans BC99
  • Preservation method of bacillus coagulans BC99
  • Preservation method of bacillus coagulans BC99

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1 strain tolerance experiment

[0019] High temperature resistance test: Take 2g of bacteria powder and spread it evenly in a 9mm petri dish, cover the dish, heat it in an oven at 120°C and 150°C respectively, take out the sample after heating for 15min and 30min, and detect the spores in it according to the microbial detection method Count, calculate its spore survival rate, test result is as follows:

[0020]

[0021] The spores of Bacillus coagulans BC99 still had a high survival rate after high temperature treatment at 120°C and 150°C, which indicated that Bacillus coagulans BC99 had good high temperature resistance.

[0022] Acid resistance test: get 3 sterilized 250ml triangular flasks (put into appropriate amount of small glass beads), add 99.0ml sterile water in each triangular flask (add 0.5% Tween 80 before the sterile water is sterilized), respectively Use hydrochloric acid to adjust the pH to 3.0, 2.5, and 2.0, accurately weigh 1.0g of Bacillu...

Embodiment 2

[0027] Embodiment 2 strain preservation method

[0028] The general method of preservation of conventional strains is to inoculate the strains in liquid medium for liquid expansion. After 12-24 hours of cultivation, the bacterial liquid is mixed with glycerol at a ratio of 1:1 to prepare glycerin with a final glycerol concentration of 15%-30%. Bacterial solution was stored at -20°C.

[0029] The invention provides a method for preserving bacillus coagulans BC99. Prepare PCA liquid medium, adjust the pH to 7.0, put 60ml PCA liquid medium in each eggplant bottle, add agar powder according to the content of 1.8%, seal and shake well, then sterilize at 121 degrees Celsius for 20 minutes, take out after sterilization Place it at a certain angle to make a slope. After the medium solidifies, place it in a 37°C incubator for 48 hours and take it out for use.

[0030] Take the preserved Bacillus coagulans BC99 glycerin tube, after thawing, suck 0.1ml of the glycerin bacteria solution...

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Abstract

The invention relates to a preservation method of bacillus coagulans BC99, and belongs to the technical field of microorganisms. The method comprises the following steps: preparing an eggplant bottle inclined plane by using a PCA culture medium, and performing air culture at 37 DEG C for 24-48 hours for later use; sucking glycerol bacterial liquid of bacillus coagulans BC99 by using a pipette, injecting the glycerol bacterial liquid into the inclined surface of the prepared eggplant bottle, and uniformly coating the bacterial liquid by using an inoculating needle from bottom to top; placing the eggplant bottle in a constant-temperature incubator at 40-45 DEG C, and culturing for 48-72 hours; and taking out the inclined plane, adding sterilized 10-30vol% glycerol into the eggplant bottle, completely eluting the lawn on the inclined plane, sub-packaging into sterilized cryopreservation tubes, and putting into a refrigerator for cold preservation. After being cultured by the method disclosed by the invention, the thalli are good in consistency, can keep relatively high activity after being preserved for a long time, do not have the problems of difficulty in later activation and the like, and are simple and convenient to operate.

Description

technical field [0001] The invention relates to a method for preserving bacillus coagulans BC99, belonging to the technical field of microorganisms. Background technique [0002] Bacillus coagulans (Bacillus coagulans) is a Gram-positive bacterium that can grow at a temperature of 30-50°C and a pH of 4.0-10.0, and can decompose sugars to produce L-lactic acid. It is a homolactic acid fermenting bacterium. Bacillus coagulans, as an excellent strain that not only has the nutritional and immune effects of lactic acid bacteria, but also has strong stress resistance of spores, has been widely used in food, medicine, health products and animal husbandry and aquatic products. [0003] Bacillus coagulans BC99, which has strong tolerance to high temperature, acidic conditions, gastric juice, and bile salts, is preserved in the China Microorganism Culture Collection Management Center, and the preservation number is: CGMCC No.19487. Due to the differences in strains and the influence ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04C12R1/07
CPCC12N1/20C12N1/04
Inventor 方曙光刘建明史祯晖
Owner 武汉微康益生菌研究院有限公司
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