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Preparation method of freeze-dried Bifidobacterium lactis powder

A technology of bifidobacterium lactis and freeze-dried bacterial powder, applied in the biological field, can solve the problems of limiting the development and application of bifidobacterium products, difficulty in achieving a high number of viable bacteria, high production costs, etc. The effect of a stable and large number of viable bacteria

Inactive Publication Date: 2018-12-18
江西仁仁健康产业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since bifidobacterium is an obligate anaerobic bacteria, it grows slowly and easily dies in an environment with oxygen, low acidity and bile, and the production cost is high
Therefore, it is still quite difficult to achieve high-density large-scale culture of bifidobacteria, and it is difficult to achieve a high number of viable bacteria in vitro culture, which limits the development and application of bifidobacteria products.

Method used

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  • Preparation method of freeze-dried Bifidobacterium lactis powder
  • Preparation method of freeze-dried Bifidobacterium lactis powder
  • Preparation method of freeze-dried Bifidobacterium lactis powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1 A kind of preparation method of Bifidobacterium lactis freeze-dried powder

[0041] 1) Preparation of strains for fermentation

[0042] 1.1) Streak culture: After diluting the Bifidobacterium lactis powder with sterile water, mark the basal culture medium in a plate, and culture it anaerobically at 39°C for 49 hours;

[0043] 1.2) Primary purification culture: Pick a single colony on the basic medium plate, inoculate it into a test tube containing 5mL basic medium, seal it, and place it in an incubator at a constant temperature of 39°C for anaerobic culture for 22 hours to obtain the primary purified bacteria suspension;

[0044] 1.3) Secondary purification culture: Inoculate the cultured primary bacterial suspension into a 100mL Erlenmeyer flask with basal medium at 4% inoculum volume, 80% liquid volume, seal, and keep at constant temperature in a 39°C incubator Anaerobic culture for 18 hours to obtain a secondary purified bacterial suspension;

[0045]...

Embodiment 2

[0059] The comparative experiment of embodiment 2 basal medium formula

[0060] The effect of oligosaccharides in the growth factors of Bifidobacterium lactis on the number of bacteria was investigated by single factor. Most oligosaccharides are not absorbed by the digestive tract, but are used by some probiotics in the large intestine, so they have unique physiological functions, can promote the growth of bifidobacteria, and effectively increase the number of viable bifidobacteria.

[0061] Table 1 Comparative experiment of Bifidobacterium lactis basal medium

[0062]

[0063] The basal medium was prepared according to the formulas of groups 1 to 4 above, and the balance was purified water. According to the inoculum amount of 4%, inoculate the cultured primary bacterial suspension into a 100 mL basal medium Erlenmeyer flask, seal it, and place it in a 39°C incubator for 18 hours of anaerobic cultivation. Calculate the yield and number of live bacteria of Bifidobacterium ...

Embodiment 3

[0064] The comparative experiment of embodiment 3 optimization culture medium formula

[0065] Table 2 Contrast experiment of Bifidobacterium lactis optimized medium

[0066]

[0067] According to the above six formulas Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ, Ⅵ, the optimized medium was prepared respectively, and the balance was sterile water. According to the inoculation amount of 5%, inoculate the bacterial suspension of the primary fermentation of the strain into the fermenter equipped with the optimized medium, the liquid volume of the optimized medium accounts for 80% of the volume of the fermenter, and cultivate at a constant temperature of 39°C for 8 hours; The stirring speed is 80rpm, and the ventilation rate is 0. After 3.5 hours of fermentation, the constant pH of the bacterial liquid is maintained at 5.6±0.5 by feeding food-grade NaOH solution. Calculate the yield and number of live bacteria of Bifidobacterium lactis. It can be known from Table 2 that formula II is the optimized medium...

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Abstract

The invention discloses a preparation method of freeze-dried Bifidobacterium lactis powder. A basic medium adopted in the invention comprises yeast peptones, yeast extract, glucose, fructo-oligosaccharide, L-malic acid and purified water, and has a pH value of 7.2; and an optimized medium includes yeast peptones, yeast extract, glucose, fructo-oligosaccharide, L-malic acid, lactose, calcium chloride and purified water, and has a pH value of 7.2, and the number of live bacteria in a fermentation broth can reach 5.53 * 10<9> cfu / ml or more. A freeze-dried bacterium powder protection agent comprises trehalose, skim milk powder, sucrose, ascorbic acid, glucose and purified water, and the total number of viable bacteria in the free-dried Bifidobacterium lactis powder can reach 9.0 * 10<11> cfu / g or more. The preparation method can effectively achieve the purposes of high-density culture of Bifidobacterium lactis, a high bacterium yield and a large number of variable bacteria in the freeze-dried powder, and facilitates realization of large-scale industrial production.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a preparation method of freeze-dried bacteria powder of Bifidobacterium lactis. Background technique [0002] Bifidobacterium lactis belongs to the genus Bifidobacterium. Bifidobacterium is a beneficial bacterium that colonizes the human intestinal tract and is the dominant flora in the intestinal tract of healthy people. Bifidobacteria belong to the Actinomycetes family, Gram-positive, aflagellate, immobile, and anaerobic. Bifidobacteria mainly colonize the ileum and colon to form a biological barrier, and have various biological functions such as regulating the function of intestinal microecological flora, biological antagonism, nutrition, regulating human immune function, and anti-tumor effect. The health of the digestive tract has the potential to prevent and treat disease. Since bifidobacteria are obligate anaerobic bacteria, they grow slowly and easily die in an environment w...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04C12R1/01
CPCC12N1/04C12N1/20
Inventor 闵祥博张春宇余萍王聪耀
Owner 江西仁仁健康产业有限公司
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