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Lactic acid bacteria freeze-drying protecting agent

A technology of freeze-drying protective agent and cryoprotectant, which is applied in the direction of preservation of microorganisms, microorganisms, biochemical equipment and methods, etc., and can solve problems such as cell killing, dynamic balance disruption, and changes in cell membrane permeability

Inactive Publication Date: 2015-08-12
大庆固元本草汉方生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, after most bacteria are freeze-dried in an unprotected state, the bacteria will be mechanically damaged, the permeability of the cell membrane will change, the dynamic balance will be disrupted, and the cells will be killed.

Method used

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  • Lactic acid bacteria freeze-drying protecting agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: preparation of lactic acid bacteria thalline

[0026] 1) Recovery of lactic acid bacteria

[0027] Take the frozen liquid of lactic acid bacteria (Lactobacillus reuteri) stored in the -80°C refrigerator for recovery, spread 2-10 μL of culture medium on each solid MRS plate, and culture at a constant temperature of 37°C for 24 hours.

[0028] 2) Subculture of lactic acid bacteria

[0029] Take the revived lactic acid bacteria (Lactobacillus reuteri) for subculture, pick a single colony on the plate and inoculate it into a 5-10mL test tube liquid MRS medium, seal it, and culture it at a constant temperature of 37°C for 12-20h. Afterwards, the bacterial solution was inoculated into the shake flask liquid MRS medium according to the inoculum amount of 2-10%, and cultured with constant temperature shaking at 37°C for 7-16h.

[0030] 3) Determination of the number of viable bacteria in the bacterial liquid

[0031] Take a certain volume of the bacterial solut...

Embodiment 2

[0035] Embodiment 2: the preparation of lactic acid bacteria freeze-dried powder

[0036] 1) Protective agent formula: skimmed milk powder with a mass concentration of 8-25wt.%.

[0037] 2) Pre-freezing: 8-25wt.% skimmed milk powder is used as a protective agent and mixed with the bacteria in proportion, so that the bacteria account for 10-50% of the total volume after mixing. Take a small amount of the mixture and pour it into a sterile plate, covering it thinly Just one layer, cover and seal with plastic wrap, put it in a -80°C refrigerator for pre-freezing, and keep it for more than 5 hours.

[0038] 3) Freeze-drying: Take out the pre-frozen plate containing the mixture of lactic acid bacteria and protective agent from the -80°C refrigerator, place it in a pre-cooled freeze dryer, set the vacuum between 5-30Pa, and freeze Keep it dry for 20-30 hours, collect the dry powder, and store it in sealed packages.

Embodiment 3

[0039] Embodiment 3: the preparation of lactic acid bacteria freeze-dried powder

[0040] 1) Protectant formula: skimmed milk powder with a mass concentration of 8-25wt.%, ectoine solution with a mass concentration of 0.5-2wt.%. The cryoprotectant is a mixture of the two in proportion. 5-30wt.% of the cryoprotectant, ectoine accounted for 0.08-0.5wt.% of the cryoprotectant;

[0041] 2) Pre-freezing: first prepare skimmed milk powder with a mass concentration of 8-25wt.%, then add 0.5-2wt.% ectoine to it to make a freeze-drying protective agent, and then mix it with bacteria in a certain volume ratio Mix so that the volume of the bacteria accounts for 10-50% of the total volume of the mixed solution, and the mass concentration of ectoine in the mixed solution is 0.05-0.5%, draw a small amount of the mixed solution and pour it into a sterile plate, covering a thin layer That’s it, cover and seal with plastic wrap, put it in a -80°C refrigerator for pre-freezing, and keep it...

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Abstract

The invention discloses a lactic acid bacteria freeze-drying protecting agent and application thereof. The protecting agent is composed of a skimmed milk powder solution and an ectoine solution, content of skimmed milk powder is 5-30wt% of the protecting agent, and content of ectoine is 0.08-0.5wt% of the protecting agent. Different from conventional protecting agents having complex formulas, the freeze-drying protecting agent is simple in formula composition, and bacteria activity after freeze-drying is higher than that of other commonly-used protecting agents, so that viable count is guarantee. Experimental study finds that the freeze-drying protecting agent plays an unexpected role in protecting lactic acid bacteria, the lactic acid bacteria after being mixed with the protecting agent are freeze-dried, obtained bacterial powder is high in activity and stability, and freeze-drying has little impact on the lactic acid bacteria, so that long-time storage and transportation of the bacterial powder are facilitated.

Description

technical field [0001] The invention relates to a novel freeze-drying protectant, in particular to a protectant for freeze-drying lactic acid bacteria. Background technique [0002] Lactic acid bacteria (LAB) is a general term for bacteria that can produce large amounts of lactic acid from fermentable carbohydrates. These bacteria are widely distributed in nature and have rich species diversity. [0003] Lactic acid bacteria can not only improve the nutritional value of food, improve food flavor, improve food preservation and added value, but also the special physiological activity and nutritional function of lactic acid bacteria are increasingly attracting people's attention. A large number of studies have shown that lactic acid bacteria can regulate the normal flora of the gastrointestinal tract of the body, maintain micro-ecological balance, improve food digestibility and biological value, reduce serum cholesterol, control endotoxins, inhibit the growth and reproduction ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12R1/225
CPCC12N1/04
Inventor 张杰刘云成卞辑金龙毕首超孙润生范东东
Owner 大庆固元本草汉方生物科技有限公司
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