Culture method for improving freeze-drying stress resistance of bifidobacterium and application thereof

A technology of bifidobacteria and freeze-dried bacteria powder, which is applied in the field of fermentation, freeze-drying, and microorganisms, can solve the problems of single stress resistance method, insignificant effect, and reduce the growth rate, survival rate, and metabolic activity of lactic acid bacteria, and achieve Improve the survival rate of freeze-drying, the effect of improving the survival rate

Active Publication Date: 2021-08-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to solve problems such as the single method for improving the freeze-drying stress resistance of bacterial strains in the prior art, the effect is not significant and how to improve the freeze-drying stress resistance of bifidobacteria by controlling fermentation conditions, the present invention is based on the following principles: high osmotic stress will Reduce the growth rate, survival rate and metabolic activity of lactic acid bacteria

Method used

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  • Culture method for improving freeze-drying stress resistance of bifidobacterium and application thereof
  • Culture method for improving freeze-drying stress resistance of bifidobacterium and application thereof
  • Culture method for improving freeze-drying stress resistance of bifidobacterium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1: fermentation and freeze-drying process of bifidobacterium

[0060] Specific steps are as follows:

[0061] (1) Preparation of bifidobacterium seed liquid:

[0062] Streak the preserved bifidobacterium on the MRS solid medium, culture at 37°C for 36 hours to obtain a single colony; pick a single colony and inoculate it in the MRS liquid medium, culture at 37°C for 24 hours to obtain the seed solution;

[0063] (2) Inoculate the prepared seed solution into a Erlenmeyer flask containing MRS liquid medium at an inoculum size of 5% (v / v), and culture it anaerobically at 37° C. for 24 hours to obtain a culture solution;

[0064] (3) Preparation of fermentation medium containing compatible solutes

[0065] Yeast extract powder FM803 24g / L, anhydrous glucose 60g / L, Tween 80 1mL / L, cysteine ​​hydrochloride 1g / L, MgSO 4 ·7H 2 O 0.35g / L, compatible solute of 10-30g / L; put the fermentation medium in a 5L fermenter, sterilize at 115°C for 15min, fill with nitrogen ...

Embodiment 2

[0071] Embodiment 2: Fermentation and lyophilization of Bifidobacterium longum subsp.infantis CCFM 687

[0072] Specific steps are as follows:

[0073] Scheme 1: The specific implementation method is the same as in Example 1, the difference is that the compatible solute in step (3) is adjusted to 20g / L anhydrous glucose, and Bifidobacterium longum subsp.infantis CCFM 687 bacteria powder is prepared, and the bacteria harvested is detected Glucose content in culture medium, osmotic pressure value at the end of fermentation and freeze-dried survival rate.

[0074] Scheme two: the specific implementation method is the same as in Example 1, the difference is that the compatible solute in step (3) is adjusted to 20g / L of glutamic acid to prepare Bifidobacterium longum subsp.infantis CCFM 687 bacterial powder, and detect the fermentation Glucose content at the end point, osmotic pressure value at the end point of fermentation and freeze-drying survival rate.

[0075] Scheme three: ...

Embodiment 3

[0092] Embodiment 3: Fermentation and lyophilization of Bifidobacterium longum subsp.longum CCFM 1102

[0093] On the basis of Example 2, the Bifidobacterium longum subsp.infantis CCFM687 of Example 2 was replaced by Bifidobacterium longum subsp.longum CCFM 1102, and the experimental results are shown in Table 2.

[0094] Table 2: Bifidobacterium longum subsp.longum CCFM 1102 fermentation and freeze-drying results using the method of this example

[0095]

[0096]

[0097] The result shows that when the cultivation ends, the glucose content of the fermented liquid is all ≥ 16.00g / L, indicating that the glucose content in the fermentation medium of the present invention is sufficient; the osmotic pressure of the fermented liquid is all close to fully suppressing the osmotic pressure at the end of the fermentation; compared with the blank group, the experimental group After freeze-drying, the number of viable bacteria and the survival rate of freeze-drying were significant...

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Abstract

The invention discloses a culture method for improving freeze-drying stress resistance of bifidobacterium and application thereof, and belongs to the technical field of microorganisms and the technical field of fermentation and freeze-drying. The invention provides the method for increasing the freeze-drying survival rate of the bifidobacterium by adding the compatible solute into the culture medium for high-density fermentation of the bifidobacterium, the bifidobacterium is inoculated into the culture medium containing the compatible solute for high-density culture to a stable phase, the freeze-drying stress resistance of the bifidobacterium can be remarkably improved, and the freeze-drying survival rate of the bifidobacterium is maximally increased by 3.02 times (under the action of the same protective agent); according to the culture method, the freeze-drying survival rate of the bifidobacteria can be remarkably increased, and the freeze-drying survival rate of the bifidobacteria is increased from 10%-20% to 40%-60%.

Description

technical field [0001] The invention relates to a cultivation method for improving the freeze-drying stress resistance of bifidobacteria and an application thereof, belonging to the technical field of microbes and the technical fields of fermentation and freeze-drying. Background technique [0002] Bifidobacterium longum is a Gram-positive, catalase-negative rod-shaped bacterium that exists in the human gastrointestinal tract, including Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium adolescentis, Bifidobacterium bifidobacterium Bifidobacteria and other 32 species. Bifidobacterium is a micro-aerobic tolerant anaerobic bacterium that is thought to reside first in the gastrointestinal tract of infants. Bifidobacteria generally form white, smooth convex colonies when grown anaerobically on solid media. Although bifidobacteria are less abundant in the gastrointestinal tract of adults, they are considered part of the gut microbiota and widely distributed in the h...

Claims

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Application Information

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IPC IPC(8): C12N1/04C12N1/20C12R1/01
CPCC12N1/04C12N1/20
Inventor 崔树茂高欣伟唐鑫毛丙永赵建新
Owner JIANGNAN UNIV
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