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Microbial freezing drying protective agent and application thereof

A dry protective agent and drying technology, which is applied in the field of microbial frozen storage, can solve the problems of complex formula of protective agent and poor stability of protective effect, and achieve the effect of solving large differences in protective effect and facilitating stability

Active Publication Date: 2010-02-17
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently no applications of stachyose as a lyoprotectant
At the same time, the protective agents commonly used in the past also have problems such as very complicated formulas of protective agents and poor stability of protective effects.

Method used

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  • Microbial freezing drying protective agent and application thereof
  • Microbial freezing drying protective agent and application thereof
  • Microbial freezing drying protective agent and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] In the study and design of protective agents, Bifidobacterium breve BB 2 For the tested strains, stachyose single factor test was carried out, in which the proportions of the other three compositions were fixed: 0.1% sodium glutamate, 0.4% galactose, and 8% skimmed milk powder. The results are shown in Table 2.

[0042] Table 2 The freeze-dried survival rate and multiple comparison results of different levels of stachyose

[0043]

[0044] Note: The uppercase and lowercase letters marked on the right side of the numbers represent the multiple comparison results with statistical significance levels p≤0.01 and p≤0.05, respectively.

[0045] It can be seen from the table that adding 1% stachyose to the protective agent can significantly improve the protection against Bifidobacterium BB. 2 freeze-drying protection rate. With the increase of stachyose content, the survival rate was also significantly improved, and the freeze-drying protection rate was strongly positive...

Embodiment 2

[0059] Culture of Bifidobacterium breve (Bifidobacterium breve BB) under suitable conditions 2 ). This fermented liquid 2000rpm (promptly per minute 2000 turns) centrifugation 5min (promptly 5 minutes), abandon supernatant, stay bacterium, get a certain amount of normal saline washing, 5000rpm centrifugal 4min, abandon supernatant, stay bacterium (washedbacteria mire, WBM), add protective agent A (0.8% stachyose, 0.1% sodium glutamate, 1.2% galactose, 12% skimmed milk powder) to 1.5 times the volume of the bacteria slime, mix well, equilibrate in a water bath at 33°C for 35 minutes, and freeze in liquid nitrogen for 20 seconds ("s" stands for "second", the same below), freeze-dried for 16 hours (h stands for hour), immediately rehydrated, 37 ° C water bath for 30 minutes, mixed, serially diluted and counted, and the survival rate was calculated, and the protection rate of the protective agent was 48.32 %. After 3 months of storage at room temperature, the same rehydration st...

Embodiment 3

[0062] Cultivate Bifidobacterium breve (Bifidobacterium breve BB) according to the method in embodiment 2 2 ). The fermented liquid was centrifuged at 3500rpm for 8min, the supernatant was discarded, and the thallus was kept, washed with a certain amount of normal saline, centrifuged at 3500rpm for 5min, the supernatant was discarded, the thallus was kept, and the protective agent B (1.5% stachyose) was added to the 1.5 times volume of the bacterium slime. , sodium glutamate 0.2%, galactose 0.6%, skimmed milk powder 12%), mix well, balance in 35°C water bath for 40min, quick freeze in liquid nitrogen for 40s, freeze dry for 15h, immediately rehydrate, 37°C water bath for 30min, mix well, continuous Dilute and count, and calculate the survival rate, and the protection rate of the protective agent is 62.94%. After 3 months of storage at room temperature, the same rehydration steps were used to calculate the survival rate, and the protection rate of the protective agent was 49.2...

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Abstract

The invention provides a microbial freezing drying protective agent using stachyose as one of main components in the protective agent. The protective agent consists of 0.8-3.5% of stachyose, 0.1-0.6%of sodium glutamate, 0.4-1.5% of galactose and 6-15% of skim milk powder. In the invention, the stachyose is applied to freeze, dry and protect strains and used as one of components of a freeze-dryingprotective agent so as to widen the application of the stachyose. Advantages of the stachyose, sodium glutamate and the like are mutually complemental to form favorable combinations of the freeze-drying protective agent. The invention has higher freeze-drying protective efficiency and stable protective effect to various bifidobacterium such as bifidobacterium breve, bifidobacterium adolesentis and the like and various lactobacillus such as lactic acid bacillus, lactococcus and the like. The invention solves the problems that freeze-drying protective agent is complicated, and the differences of protective effects are great in the study of microbial formulations and provides favorable compositions of the protective agent for the preservation of probiotics such as bifidobacterium, lactobacillus, saccharomycetes and the like, thereby being beneficial to preserving the strain stably.

Description

technical field [0001] The invention relates to microorganism freezing storage technology, in particular to a protective agent for freeze-drying preservation of microorganisms, and also relates to the use of the protective agent. Background technique [0002] In recent years, due to the needs of food safety, the research and development of microecological preparations has developed rapidly. Bifidobacteria and lactic acid bacteria are favored in the research of microecological preparations, except that they have the physiological functions of common probiotics such as: promoting In addition to the absorption and utilization of nutrients, biological barrier and biological antagonism, and immune regulation, it also has the effects of inhibiting endotoxin production, improving the breeding environment, and producing bacteriocins with broad-spectrum antibacterial effects. But it is rarely used in actual production. The reason is that: bifidobacteria and lactic acid bacteria have...

Claims

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Application Information

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IPC IPC(8): C12N1/04C12N1/20C12N1/16C12R1/01C12R1/225C12R1/245C12R1/23C12R1/25C12R1/645C12R1/72C12R1/865
Inventor 张日俊梁晓明
Owner CHINA AGRI UNIV
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