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Method for improving lyophilized activity of Oenococcus oeni by utilizing stress pretreatment

An oenophilus and pretreatment technology, which is applied in microorganism-based methods, wine preparation, alcoholic beverage preparation, etc., can solve the problems of harsh and complex wine habitat, reduction of surviving bacteria, bacterial damage, etc., and achieve strong industrialization. Implementability, increase freeze-drying survival rate, increase the effect of taste and flavor

Inactive Publication Date: 2012-07-11
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] In addition, the habitat of the wine after alcohol fermentation is harsh and complex. In the case of direct manual inoculation, it will often cause a large number of death of the bacteria, and the MLF cannot be successfully activated.
According to relevant literature reports, there are two reasons for this. One is that the bacteria obtained in the laboratory culture lost their natural resistance to the wine environment, resulting in a decrease in the survival rate of the inoculation; the other was that the bacteria were damaged during the freeze-drying process, resulting in The number of surviving bacteria decreased after freeze-drying

Method used

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  • Method for improving lyophilized activity of Oenococcus oeni by utilizing stress pretreatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Centrifuge 1000ml of Oenococcus oeni cells grown to the stable stage at 6000×g for 10min at 4°C, discard the supernatant, wash with normal saline and collect the sludge, add 100ml of ATB medium to the sludge, Adjust the pH value of the ATB medium to 3.5 with hydrochloric acid, mix thoroughly, and let stand at 25°C for 2h.

[0036] After the stress-treated thalline was centrifuged under the same conditions, the bacteria sludge was collected after washing with normal saline, and 100ml of sodium glutamate solution with a mass concentration of 2.5% was added to the bacteria sludge, mixed evenly, and then placed at room temperature for 20 minutes Freeze-dry in a vacuum freeze-dryer for 28 hours to obtain freeze-dried bacterial powder.

[0037] After adding 100ml of ATB medium to the freeze-dried bacteria powder to rehydrate, count the number of live bacteria by plate colony counting method. In the freeze-dried bacteria powder after ordinary freeze-drying, the number of live ...

Embodiment 2

[0040] Centrifuge 1000ml of Oenococcus oeni thallus grown to the stable stage at 3000×g for 15min at 4°C, discard the supernatant, wash with normal saline and collect the sludge, add 100ml of ethanol with a concentration of 8% ATB medium, mix thoroughly, and stand at 25°C for 2.5h.

[0041] After the stress-treated thalline was centrifuged under the same conditions, the bacteria sludge was collected after washing with normal saline, and 50 ml of sodium glutamate solution with a mass concentration of 2.5% was added to the bacteria sludge, mixed evenly, and left to stand at room temperature for 30 minutes. Freeze-dry in a vacuum freeze-dryer for 25 hours to obtain freeze-dried bacterial powder.

[0042] After adding 50ml of ATB medium to the freeze-dried bacteria powder and rehydrating, count the number of live bacteria by plate colony counting method. 10 cfu / ml, and the number of live bacteria in the freeze-dried bacterial powder was 2.55×10 after the ATB medium stress of 8% e...

Embodiment 3

[0045] Centrifuge 500ml of Oenococcus oeni cells grown to the stable stage at 3000×g, 4°C for 10min, discard the supernatant, wash with normal saline and collect the sludge, add 100ml of ethanol concentration of 10% to the sludge ATB medium, mix thoroughly, and stand at 25°C for 2h.

[0046] Centrifuge the stress-treated bacteria under the same conditions, wash with normal saline, collect the bacteria sludge, add 50ml of 5% sodium glutamate solution, mix well, let stand at room temperature for 30min, and freeze in a vacuum freeze dryer. Dry for 25 hours to obtain freeze-dried bacteria powder.

[0047] After adding 50ml of ATB medium to the freeze-dried bacteria powder and rehydrating, the number of viable bacteria was counted by the plate colony counting method. 10 cfu / ml, the number of viable bacteria was 1.3×10 after 2 hours of stress in ATB medium with ethanol concentration of 10%. 10 cfu / ml.

[0048] Put the freeze-dried bacteria powder into 1000ml of wine after alcohol...

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Abstract

The invention relates to a method for improving lyophilized activity of Oenococcus oeni by utilizing stress pretreatment, which comprises the following steps: performing enrichment culture of Oenococcus oeni strain to reach the stable period, centrifugally collecting the bacterial sludge, resuspending in the culture medium ATB containing the stress treatment fluid, standing at 25 DEG C for 2-3h, centrifuging, harvesting bacterial body, adding proper sodium pyruvate solution with mass concentration of 2-5% as the lyophilized protective agent and performing freeze drying to obtain lyophilized bacteria powder. The lyophilized bacteria powder prepared by the method of the invention can be used as direct Oenococcus oeni fermentation starter. The method of the invention introduces the idea of stress pretreatment, and uses sodium pyruvate as the lyophilized protective agent, which is the main component of the monosodium glutamate and is safe and no harm to human and poultry and has various health-care effects, the introduced idea can be used in the production process of direct edible fermentation starter.

Description

technical field [0001] The invention belongs to the technical field of production of wine freeze-dried starter and lactic acid bacteria products, and in particular relates to a method for improving the freeze-dried activity of Oenococcus oeni by stress pretreatment. Background technique [0002] In recent years, my country's wine production has maintained a strong growth momentum, but the malic acid-lactic acid starter used in the wine industry is all dependent on foreign imports, and the cost is as high as about 60 million yuan per year. With the increase of wine production year by year, this Item costs will still be further expanded. In order to effectively control malolactic fermentation (MLF), the production and use of direct-throwing lactic acid bacteria starter has attracted extensive attention of wine microbiologists. [0003] Direct throw starter refers to a safe and efficient biological product with strong fermentation activity and high number of viable bacteria, wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12G1/022C12R1/01
Inventor 樊明涛张国强
Owner NORTHWEST A & F UNIV
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