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Probiotic agent capable of improving strain storage stability as well as preparation method and application of probiotic agent

A technology of storage stability and probiotics, applied in the field of microorganisms, can solve the problems that the bacterial agent of the composition does not achieve the expected effect, it is difficult to ensure the basic activity, and it is difficult to isolate exogenous stress, so as to ensure the soundness of the cell structure and physiological functions active effect

Pending Publication Date: 2022-07-29
JIANGSU WECARE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, judging from the actual industrial application and the feedback of data results, many composition bacterial agents have not achieved the expected effect, especially for different levels of active stress interventions under different environmental conditions, it is difficult to guarantee their basic activity, especially for strict Oxygenous intestinal microbial strains, conventional theoretical "encapsulation" means are difficult to isolate exogenous stress, and further improve or protect the survival rate of different types of active cell substances under long-term room temperature (or high temperature) conditions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Preparation of freeze-dried particles of Bifidobacterium bifidum BBi32

[0070] S1. Inoculate Bifidobacterium bifidum (the preservation number is CGMCC No. 16923) with 3.0% inoculum in MRS medium for activation and expansion (temperature is 34°C, time is 20h), and then fermentative culture (inoculum amount is 20h). 2.0%, temperature 34°C, time 14h), centrifuge (6500rpm, 30min) after fermentation, pour out the supernatant to obtain Bifidobacterium bifidum BBi32 concentrated bacterial suspension (containing Bifidobacterium bifidum BBi32 ≥1×10 11 CFU / g);

[0071] S2, uniformly mix the protective agent and the concentrated bacterial suspension according to the mass ratio of concentrated bacterial suspension: protective agent=1:0.5, and equilibrate at 20° C. for 20 minutes to obtain an active mixture. Among them, the protective agent contains 55% of inactivated lactic acid bacteria cells, 2.0% of the cell membrane stabilizer (the mass ratio of selenocysteine ​​and mannitol...

Embodiment 2

[0076] Preparation of Mucinophilic-Akermansia (AKK) lyophilized granules

[0077] S1. Inoculate Akkermansia muciniphila (ATCC BBA-835) with 5% inoculum in the medium (add 0.1% mucin to the commercially available BHI brain heart infusion medium) for activation and expansion Incubate (temperature is 37°C, time is 20h, 5% CO 2 , 95%N 2 , pH 6.0), then carry out fermentation culture (inoculum size 2.0%, temperature 37°C, time 18h), centrifuge (rotation speed 12000rpm, time 15min) after fermentation to obtain AKK concentrated bacterial suspension (containing AKK≥1× 10 8 CFU / g), due to the characteristics of the bacteria, anaerobic protection is required for the whole process of cultivation, and the protective gas used is nitrogen / carbon dioxide mixed gas.

[0078] S2. The inactivated lactic acid bacteria cells and the cell membrane stabilizer are used for compound combination to form a basic protection formula, which is uniformly mixed with the concentrated bacterial suspension ...

Embodiment 3

[0083] This example provides a preparation method for Bifidobacterium bifidum BBi32 freeze-dried particles, which differs from Example 1 only in that the cell membrane stabilizers "selenocysteine ​​and mannitol" are replaced with the same amount of " Mannitol", other reference to Example 1.

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PUM

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Abstract

The invention provides a probiotic agent capable of improving the storage stability of a strain as well as a preparation method and application of the probiotic agent. The probiotic agent capable of improving the storage stability of the strain is prepared from the following raw materials: probiotics, a protective agent and an embedding medium, the protective agent comprises inactivated lactic acid bacteria and a cell membrane homeostasis agent; the cell membrane homeostasis agent comprises any one of selenocysteine, methionine, mannitol or phosphatidylserine or a combination of at least two of the selenocysteine, the methionine, the mannitol and the phosphatidylserine; the embedding medium comprises a polylactic acid-glycolic acid copolymer. The protective agent is mainly used for protecting the endogenous steady state of probiotic cells, and the embedding agent is used for forming a micro-capsule structure, isolating adverse exogenous conditions, further protecting the probiotics and improving the storage stability of the probiotics. Compared with the prior art, the scheme provided by the invention has a better effect of improving the storage stability of the strain, is not only suitable for common probiotics such as lactic acid bacteria, but also suitable for strict anaerobic strains such as Ackerman's bacteria, and has an important application value.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to a probiotic agent for improving the storage stability of bacterial strains and a preparation method and application thereof. Background technique [0002] The storage stability of probiotic products such as lactic acid bacteria at different temperatures has been a core issue that has plagued the development of the industry for a long time, resulting in product implementation standards, regulatory certification and the process of host taking due to the attenuation of lactic acid bacteria activity. There are many patent documents claiming to have effective protection methods for lactic acid bacteria products under normal temperature or worse conditions, usually by reducing the water activity (aw) in the environment or enhancing the antioxidant system, micro-encapsulation, etc. means to meet its ends. [0003] CN102325873A discloses the preparation of lactic acid bacteria conc...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N11/096A61K9/50A61K35/741A61K47/46A61K47/34A61P1/00A23C9/123A23L33/135C12R1/225C12R1/46C12R1/01C12R1/25C12R1/245C12R1/23
CPCC12N1/20C12N11/096A61K35/741A61K47/46A61K9/5031A61P1/00A23L33/135A23C9/1234A23C9/1236A23C9/1238A23V2002/00A23V2400/145A23V2400/151A23V2400/137A23V2400/143A23V2400/181A23V2400/165A23V2400/173A23V2400/231A23V2400/413A23V2400/427A23V2400/125A23V2400/513A23V2400/529A23V2400/517A23V2400/519A23V2400/175A23V2400/113A23V2400/169A23V2400/531A23V2400/533A23V2400/249A23V2200/3204
Inventor 方曙光陈珂可冯胜鹏严涛杨恒飞程文吴嘉宝
Owner JIANGSU WECARE BIOTECHNOLOGY CO LTD
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