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Anaerobic ultralow temperature strain preserving agent

An anaerobic ultra-low temperature, strain preservation technology, applied in the field of strain preservatives, can solve the problems of strict anaerobic microbial life activity inhibition, difficult to reach laboratories, oxygen and miscellaneous bacteria contamination, etc., to achieve stable and increased protection effect. The scope of use and the effect of solving preservation problems

Active Publication Date: 2015-02-25
NORTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the requirements for this technical equipment are high, and it is difficult for general laboratories to achieve
In addition, in the preparation process before freeze-drying, the contamination of oxygen and bacteria will inevitably be introduced, which will inhibit the life activities of strictly anaerobic microorganisms, and even die

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Anaerobic ultra-low temperature preservation agent, which is composed of the following raw materials in volume percentage (ml / ml): 20.8% nutrient broth, 26.3% protective agent, 31.5% dispersant and 21.4% deionized water.

[0026] in:

[0027] The nutrient broth is made by mixing 0.3% polypeptide substance-yeast extract powder, 1.0% protein substance-fish meal, and 98.7% deionized water by mass percentage (g / g).

[0028] The protective agent is composed of 35.0% alcohols - isopropanol, anhydrous alcohol obtained after blast drying at 80.0°C overnight, 40.0% antifreeze - sulfoxide, 25.0% fatty acids - linoleic acid by mass Percentage (g / g) mixed.

[0029] The dispersant is composed of 66.8% protein - whole milk, 32.4% sugar - maltose, 0.5% sodium glutamate, 0.3% L-cysteine ​​mixed according to mass percentage (g / g) become.

[0030] The deionized water is oxygen-free deionized water, and its resistance value is 18.0 megohms.

[0031] The preparation meth...

Embodiment 2

[0041] Example 2 Anaerobic ultra-low temperature preservation agent, which is composed of the following raw materials in volume percentage (ml / ml): 20.0% nutrient broth, 29.8% protective agent, 30.3% dispersant and 19.9% ​​deionized water.

[0042] in:

[0043] The nutrient broth is made by mixing 0.5% polypeptide substance-beef extract powder, 1.5% protein substance-peptone, and 98.0% deionized water by mass percentage (g / g).

[0044] The protective agent is composed of 40.0% alcohols—glycerin, anhydrous alcohol obtained by blast drying at 80.0°C overnight, 40.0% antifreeze—dimethyl sulfoxide, 20.0% fatty acids—conjugated Linoleic acid is mixed by mass percentage (g / g).

[0045] The dispersant is composed of 68.0% protein - skimmed milk, 31.4% sugar - lactose, 0.4% sodium glutamate, 0.2% L-cysteine ​​mixed by mass percentage (g / g) .

[0046] The deionized water is oxygen-free deionized water, and its resistance value is 18.0 megohms.

[0047] The preparation method ...

Embodiment 3

[0057] Example 3 Anaerobic ultra-low temperature bacteria preservative, the preservative is composed of the following raw materials in volume percentage (ml / ml): 21.1% nutrient broth, 29.0% protective agent, 30.2% dispersant and 19.7% deionized water.

[0058] in:

[0059] The nutrient broth is made by mixing 0.4% polypeptide substance-wort extract powder, 1.6% protein substance-silkworm chrysalis powder, and 98.0% deionized water according to the mass percentage (g / g).

[0060] The protective agent is composed of 40.0% alcohols - propylene glycol, anhydrous alcohol obtained after air drying at 80.0°C overnight, 35.0% antifreeze - dimethyl sulfone, 25.0% fatty acids - linolenic acid by mass percentage (g / g) mixed.

[0061] The dispersant consists of 68.0% protein - a mixture of whole milk and skimmed milk at a volume ratio of 1:1 (ml / ml), 31.55% sugar - fructose, 0.3% sodium glutamate, 0.15 % L-cysteine ​​is mixed by mass percentage (g / g).

[0062] The deionized water...

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Abstract

The invention relates to an anaerobic ultralow temperature strain preserving agent. The anaerobic ultralow temperature strain preserving agent consists of the following materials in percentage by weight: 20.0%-21.1% of nutrient broth, 26.3%-30.0% of protective agent, 30.0%-31.5% of protective agent and 19.3%-21.4% of deionized water. The invention further discloses a preparation method of the preserving agent. According to the anaerobic ultralow temperature strain preserving agent disclosed by the invention, environment conditions beneficial to microorganism dormancy are sufficiently created according to the basic principle of the strain preservation and basic metabolism level of the strain is greatly reduced; the anaerobic ultralow temperature strain preserving agent can well protect aerobic, strictly anaerobic or facultative anaerobic microorganism with stable protective effect, the strain variation degree is extremely small and the recovery rate is high; moreover, the anaerobic ultralow temperature strain preserving agent not only can be used for anaerobic ultralow temperature preservation of the strain, but also can be used for liquid ammonia (-196 DEG C) ultralow temperature preservation and low temperature freeze drying preservation of the strain, so that the using range of the preserving agent is increased, therefore, the strain preserving difficulty under the anaerobic ultralow temperature condition is solved, and the preserving period of the strain is prolonged to several tens of years.

Description

technical field [0001] The invention relates to a bacterial strain preservation agent, in particular to an anaerobic ultra-low temperature bacterial strain preservation agent. Background technique [0002] All strains with excellent traits obtained from production practice or scientific research, including wild strains and excellent mutant strains or genetically engineered strains, are important germplasm resources of the country. Therefore, strain preservation is the basis of all microbial work. The purpose of strain preservation is to keep strains from dying, mutating, and being polluted by miscellaneous bacteria after preservation, and to maintain their excellent properties, so as to facilitate the application of production and scientific research. [0003] Preservatives have a great influence on the survival rate of anaerobic cryopreservation of microbial strains. Proper selection of preservatives is the key factor to improve the survival rate of microbial anaerobic cryo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04
Inventor 牛锋吴慧昊黄晓磊龙玲李建江徐红伟傅思武关晓莉
Owner NORTHWEST UNIVERSITY FOR NATIONALITIES
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