Freeze-drying protective agent and application thereof to freeze-dried lactobacillus reuteri

A technology of Lactobacillus reuteri and freeze-drying protective agent, which is applied in the field of microorganisms, can solve the problems of low viable count of Lactobacillus reuteri, destruction of pH dynamic balance, and decreased activity of Lactobacillus reuteri, so as to improve the efficiency of freeze-drying. The effect of stem survival

Active Publication Date: 2020-09-04
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of preparing Lactobacillus reuteri freeze-dried powder, although freeze-drying has less damage to Lactobacillus reuteri, due to the process of freeze-drying, Lactobacillus reuteri is prone to low temperature stress and freezing stress damage. Changes in cell membrane permeability, protein denaturation and inactivation, and disruption of pH dynamic balance, etc., and these conditions will lead to a decrease in the activity of Lactobacillus reuteri. Add specific lyoprotectants, such as sucrose, trehalose, skim milk, hydroxyethylcellulose, polysucrose, ascorbic acid, hyaluronic acid, alcohols and / or sodium salts, etc. stem survival rate
[0005] However, there are few freeze-drying protectants specially developed for the probiotic Lactobacillus reuteri at present, and most of the effects are not ideal. For example, in the patent application text with the publication number CN101974463A, Zhu Zhanbo et al. When freeze-drying Lactobacillus reuteri with lyoprotectants of sucrose, glycerin, and sorbitol, the lyophilization survival rate is only 70%
And, due to the specificity between different bacterial strains (specific references: Huang Ke. Research progress of vacuum freeze-dried lactic acid bacteria [J]. Journal of Xi'an University of Arts and Science: Natural Science Edition, 2009 (03): 55-58.), The existing freeze-drying protectants developed for other probiotics are not ideal in terms of improving the freeze-dried survival rate of Lactobacillus reuteri. For example, in the patent application text with publication number CN106434463A, Yu When Ping et al. lyophilized Lactobacillus rhamnosus using a lyoprotectant containing skimmed milk powder, maltodextrin, trehalose, Tween-80, glycerol, and sodium glutamate, the lyophilized survival rate was 60%. However, When using the same freeze-drying protection agent to freeze-dry Lactobacillus reuteri, the freeze-drying survival rate is only 40%; the publication number is that in the patent application text of CN106047709A, people such as Cui Shumao use whey protein isolate (WPI), inulin , sorbitol and ascorbic acid freeze-dried Lactobacillus plantarum, the freeze-dried survival rate was as high as 100%, however, when the same freeze-dried Lactobacillus reuteri was freeze-dried, the freeze-dried survival rate was only 50%
The above factors all make the number of live bacteria of Lactobacillus reuteri in most existing Lactobacillus reuteri freeze-dried powder products low

Method used

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  • Freeze-drying protective agent and application thereof to freeze-dried lactobacillus reuteri

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: Preparation of Lactobacillus reuteri freeze-dried powder (Lactobacillus reuteri GDMCC No.60515)

[0039] Specific steps are as follows:

[0040](1) Configure freeze-drying protective agents 1-12 according to the formula in Table 1; among them, stachyose solution, sodium sulfate solution, maltodextrin solution, trehalose solution, Tween-80, glycerin solution, inulin solution, sorbic acid Alcohol solution and sucrose solution are sterilized at 115°C for 20 minutes, sodium caseinate solution is sterilized at 65°C for 30 minutes, glycine solution, hyaluronic acid solution, ascorbic acid solution, and sodium glutamate solution are sterilized through membrane, and skimmed milk powder solution is sterilized at 105°C Sterilize for 10 minutes, whey protein isolate (WPI) solution is sterilized at 85°C for 30 minutes;

[0041] (2) Dip the Lactobacillus reuteri GDMCC No.60515 bacterial solution in the bacteria preservation tube with an inoculation loop, streak it on ...

Embodiment 2

[0050] Embodiment 2: Preparation of Lactobacillus reuteri freeze-dried powder

[0051] Specific steps are as follows:

[0052] On the basis of Example 1, Lactobacillus reuteri GDMCC No.60515 was replaced with Lactobacillus reuteri CGMCC No.14394 to obtain Lactobacillus reuteri freeze-dried powder 13-24.

[0053] Detect the number of live bacteria of Lactobacillus reuteri in Lactobacillus reuteri freeze-dried powder 13~24, and calculate the freeze-dried survival rate of Lactobacillus reuteri in Lactobacillus reuteri freeze-dried powder 13~24 (detection The results are shown in Table 3).

[0054] As can be seen from Table 3, use the freeze-dried powder of Lactobacillus reuteri prepared by containing stachyose 120g / L, sodium caseinate 40g / L, glycine 15g / L and sodium sulfate 30g / L. The freeze-dried survival rate of Lactobacillus eiferii was the highest, as high as 104.42±2.47%.

[0055] The freeze-drying survival rate of Lactobacillus reuteri in table 3 Lactobacillus reuteri fr...

Embodiment 3

[0057] Embodiment 3: the preparation of Lactobacillus reuteri freeze-dried powder

[0058] Specific steps are as follows:

[0059] On the basis of Example 1, Lactobacillus reuteri GDMCC No.60515 was replaced with Lactobacillus reuteri GDMCC No.60735 to obtain Lactobacillus reuteri freeze-dried powder 25-36.

[0060] Detect the number of live bacteria of Lactobacillus reuteri in Lactobacillus reuteri freeze-dried powder 25~36, and calculate the freeze-dried survival rate of Lactobacillus reuteri in Lactobacillus reuteri freeze-dried powder 25~36 (detection The results are shown in Table 4).

[0061] As can be seen from Table 4, use the freeze-dried powder of Lactobacillus reuteri prepared by the freeze-dried powder containing stachyose 120g / L, sodium caseinate 40g / L, glycine 15g / L and sodium sulfate 30g / L. The freeze-dried survival rate of Lactobacillus eiferii was the highest, as high as 99.72±4.01%.

[0062] The freeze-drying survival rate of Lactobacillus reuteri in table...

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Abstract

The invention discloses a freeze-drying protective agent and application thereof to freeze-dried lactobacillus reuteri, and belongs to the technical field of microorganisms. The invention provides a freeze-drying protective agent capable of obviously improving a freeze-drying survival rate of the lactobacillus reuteri. Ingredients of the freeze-drying protective agent comprise stachyose, sodium caseinate, glycine and sodium sulfate; and when lactobacillus reuteri freeze-dried powder is prepared by using the freeze-drying protective agent, the freeze-drying survival rate of the lactobacillus reuteri can reach 100%.

Description

technical field [0001] The invention relates to a freeze-drying protectant and its application in freeze-dried Lactobacillus reuteri, belonging to the technical field of microorganisms. Background technique [0002] Lactobacillus reuteri (Lactobacillus reuteri) is one of the important probiotics, it can well adhere to the intestinal mucosa, regulate the structure of intestinal flora, prevent the human body from suffering from intestinal diseases, and it can well Improve immunity, thereby promoting human health. In addition, it can also improve allergic constitution, treat constipation in children, female diseases, etc. [0003] Due to its remarkable probiotic effect, Lactobacillus reuteri has increasingly become the focus of people's research, development and production. At present, Lactobacillus reuteri has been widely used in food, medicine and other fields. [0004] Lactobacillus reuteri freeze-dried powder is the most common Lactobacillus reuteri product on the market...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04C12R1/225
CPCC12N1/20C12N1/04
Inventor 赵建新崔树茂谭莎莎唐鑫毛丙永陆文伟翟齐啸陈卫张灏
Owner JIANGNAN UNIV
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