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Method for reducing secretion amount of lactobacillus rhamnosus surface substance

A technology of Lactobacillus rhamnosus and secretion volume, applied in the field of microorganisms, can solve the problems of pH dynamic balance damage, protein denaturation inactivation, reduction of Lactobacillus rhamnosus, etc., and achieves the effect of reducing the difficulty of collection and reducing the secretion volume.

Active Publication Date: 2020-08-25
JIANGNAN UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although freeze-drying has less damage to Lactobacillus rhamnosus, due to the process of freeze-drying, Lactobacillus rhamnosus is prone to changes in cell membrane permeability, protein denaturation and inactivation, Conditions such as the destruction of the pH dynamic balance, and these conditions will lead to a decrease in the activity of Lactobacillus rhamnosus. Therefore, before freeze-drying, it is necessary to add a specific freeze-drying protective agent, such as stachyus, to the Lactobacillus rhamnosus cells. sugar, trehalose, inulin and / or isomaltooligosaccharide, etc.
[0006] And, since Lactobacillus rhamnosus can produce surface substances (mainly comprising capsular polysaccharides and surface proteins) during growth and metabolism, the presence of surface substances will reduce the lyophilization rate of Lactobacillus rhamnosus cells during the freeze-drying process. Survival rate (see references for details: Wu Sheng, Cui Shumao, Mao Bingyong, Tang Xin, Zhao Jianxin, Zhang Hao, Chen Wei. Effect of surface substances of Lactobacillus rhamnosus on its freeze-drying survival rate[J / OL]. Food and Fermentation industry: 1-9), and, the existence of surface material can reduce the live bacteria number that can unit mass Lactobacillus rhamnosus bacterium powder, in addition, surface material contains polyhydroxyl structure, and polyhydroxyl structure is easy to combine with water molecule, and then As a result, it is more difficult to collect Lactobacillus rhamnosus cells in the culture medium of Lactobacillus rhamnosus. Therefore, before freeze-drying, it is also necessary to reduce the secretion of surface substances of Lactobacillus rhamnosus as much as possible.
[0007] However, at this stage, there is no good method for reducing the secretion of surface substances of Lactobacillus rhamnosus. Therefore, it is urgent to find a method for reducing the secretion of surface substances of Lactobacillus rhamnosus to improve the growth rate of Lactobacillus rhamnosus. Freeze-drying survival rate in the freeze-drying process, increase the number of live bacteria per unit mass of Lactobacillus rhamnosus powder, and reduce the difficulty of collecting Lactobacillus rhamnosus cells in the culture medium of Lactobacillus rhamnosus

Method used

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  • Method for reducing secretion amount of lactobacillus rhamnosus surface substance
  • Method for reducing secretion amount of lactobacillus rhamnosus surface substance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: the preparation of Lactobacillus rhamnosus bacterial powder

[0052] Specific steps are as follows:

[0053] Dip the Lactobacillus rhamnosus CCFM1060 bacteria liquid in the bacteria preservation tube with an inoculation loop, streak it on the MRS solid medium, and culture it at 37°C for 36 hours to obtain a single colony; pick a single colony and inoculate it in the MRS liquid medium, 37 Cultivate at a constant temperature for 12 hours at ℃ to obtain a seed liquid; inoculate the seed liquid into medium A at an inoculation amount of 2% (v / v), and culture at a constant temperature of 37°C and a pH of 5 for 6 hours, 9 hours, and 12 hours respectively to obtain a cultured Liquid 1-3; Centrifuge culture liquid 1-3, collect bacteria 1-3;

[0054] Among them, the composition of medium A is: arabinose 20g / L, yeast extract powder FM52825g / L, magnesium sulfate 0.58g / L, manganese sulfate 0.25g / L and Tween-801g / L.

[0055] Part of the cells 1-3 were taken and the su...

Embodiment 2

[0066] Embodiment 2: the preparation of lactobacillus rhamnosus bacterial powder

[0067] Specific steps are as follows:

[0068] On the basis of Example 1, Lactobacillus rhamnosus CCFM1060 was replaced with Lactobacillus rhamnosus CCFM1064, Lactobacillus rhamnosus CCFM1068, and Lactobacillus rhamnosus CCFM1070 respectively, to obtain cells 4-12 and rhamnose milk Bacillus powder 4-12.

[0069] Detect the surface protein content and capsular polysaccharide content on the surface of thalline 4~12 (detection results are shown in Table 3), detect the number of live bacteria of Lactobacillus rhamnosus in Lactobacillus rhamnosus bacteria powder 4~12, and calculate the The freeze-dried survival rate of Lactobacillus rhamnosus in Lactobacillus lysus bacteria powder 4-12 (test results are shown in Table 4).

[0070] It can be seen from Tables 3-4 that when cultured for 9 hours or more, the surface protein content and capsular polysaccharide content on the surface of the cells 4-12 we...

Embodiment 3

[0075] Embodiment 3: the preparation of Lactobacillus rhamnosus bacterial powder

[0076] Specific steps are as follows:

[0077] On the basis of Example 1, the carbon source in medium A was replaced with glucose, galactose, glucose fructose or sucrose, respectively, to obtain bacterial cells 13-16 and Lactobacillus rhamnosus bacterial powder 13-16.

[0078] Detect the surface protein content and capsular polysaccharide content on the surface of thalline 13~16 (detection results are shown in Table 5), detect the number of viable bacteria of Lactobacillus rhamnosus in Lactobacillus rhamnosus bacteria powder 13~16, and calculate the number of viable cells of Lactobacillus rhamnosus The freeze-dried survival rate of Lactobacillus rhamnosus in Lactobacillus lysus bacteria powder 13-16 (the test results are shown in Table 6).

[0079] From Tables 1 to 2 and Tables 5 to 6, it can be seen that the choice of carbon source has a great influence on the surface protein content and capsu...

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Abstract

The invention discloses a method for reducing secretion amount of lactobacillus rhamnosus surface substance, and belongs to the technical field of microorganisms. The invention provides a method for reducing the secretion amount of the lactobacillus rhamnosus surface substances, which can obviously reduce the secretion amount of lactobacillus rhamnosus surface substances, thereby lowering the collection difficulty of lactobacillus rhamnosus thalli in a lactobacillus rhamnosus culture solution. According to the method, lactobacillus rhamnosus is inoculated into a culture medium containing arabinose, yeast extract powder, magnesium sulfate, manganese sulfate and Tween-80 to be cultured, so that the secretion amount of capsular polysaccharide on the surface of lactobacillus rhamnosus thallusis only 1.15 * 10 <-12 > mg / CFU; and the secretion amount of the surface protein is only 1.90 * 10 <-10 > mg / CFU.

Description

technical field [0001] The invention relates to a method for reducing the secretion of surface substances of Lactobacillus rhamnosus, belonging to the technical field of microorganisms. Background technique [0002] Lactobacillus rhamnosus (Lactobacillus rhamnosus) mostly exists in the intestines of humans and animals, belongs to the genus Lactobacillus, subspecies rhamnosus, and is a Gram-positive probiotic that is anaerobic and acid-resistant and does not produce spores. Lactobacillus rhamnosus cannot utilize lactose and can ferment a variety of monosaccharides. Most strains can produce a small amount of soluble ammonia, but do not produce indole and hydrogen sulfide. [0003] Lactobacillus rhamnosus has outstanding performance in gastric acid and bile resistance, and can enter the human intestinal tract alive, while most other probiotic species have died due to the action of gastric acid and bile before entering the intestinal tract. After colonizing and multiplying in t...

Claims

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Application Information

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IPC IPC(8): C12N1/38C12N1/20C12N1/04C12R1/225
CPCC12N1/38C12N1/20C12N1/04
Inventor 赵建新崔树茂吴晟唐鑫毛丙永陆文伟翟齐啸陈卫张灏
Owner JIANGNAN UNIV
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