Protective agent for preserving strains and preservation method

A technology of protective agent and bacterial strain, which is applied in the direction of preservation of microorganisms, etc., can solve the problems of weakening of favorable traits, large demand for storage equipment, cumbersome operation, etc., and achieve the effects of eliminating unevenness, convenient search and activation, and simple preparation procedures

Pending Publication Date: 2020-07-28
江苏华绿生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, in the regular transfer method, the water in the culture medium on the slant is easy to evaporate during storage and shrink, dry up, collapse, and change color, which increases its concentration and osmotic pressure, which will cause the strain to degenerate or even die; some favorable traits after continuous passage Will weaken; easy to cause bacterial contamination
Liquid paraffin oil preservation method: paraffin wax is mostly chemically pure. If it contains impurities, especially toxic substances, it will affect the production performance of the strain when used in the preservation of the strain; during the preservation process, the paraffin wax will decrease due to evaporation, which will cause the strain to degenerate. Even death; the slope for preservation must be placed upright, which occupies too much storage space and requires a lot of storage equipment
Sand tube preservation method: The soil as one of the storage carriers should be taken from barren soil, and the requirements for materials are high. Sand and soil need to be washed, removed from impurities, and iron filings. The proportion of sand in the sand tube is not fixed. Affected by a variety of characteristics, it often needs to be determined through experiments, with low survival rate and high mutation rate
Freeze vacuum drying method: high requirements for equipment, long preparation time and high requirements for operators’ skills
Liquid nitrogen ultra-low temperature freezing method: It needs to be frozen slowly to -40°C first, and then quickly cooled to -190°C in a liquid nitrogen tank for storage. The operation is relatively cumbersome and requires high equipment requirements

Method used

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  • Protective agent for preserving strains and preservation method

Examples

Experimental program
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Effect test

Embodiment 1

[0011] Embodiment 1: adopt 3 kinds of flat mushroom bacterial strains provided by Chinese Academy of Agricultural Sciences Edible Fungi Research and Development Center to be the participating bacterial strains, be flat mushroom No. 1 strain, flat mushroom No. 2 bacterial strain, flat mushroom No. 3 bacterial strain, investigate this preservation strain Effects of protective agents on the survival rate of three strains of Pleurotus ostreatus after cryopreservation.

[0012] Preparation of protective agent: 10% glycerin, 5% trehalose solution, and the balance is deionized water.

[0013] Inoculate the mycelia of three kinds of Pleurotus ostreatus strains into PDA medium respectively, cultivate them until the mycelium is full at 25° C., adopt the preservation method of the present invention to preserve the strain protection agent, and put them into 100 ml after subpackaging in cryopreservation tubes. Well frozen storage tube box, marked on the box cover, placed in a -80 ℃ refrige...

Embodiment 2

[0014] Embodiment 2: the test bacterial strain is identical;

[0015] Preparation of protective agent: 8% glycerin, 4% trehalose solution, and the balance is deionized water.

[0016] Inoculate the mycelia of three kinds of Pleurotus ostreatus strains into PDA medium respectively, cultivate them until the mycelium is full at 25° C., adopt the preservation method of the present invention to preserve the strain protection agent, and put them into 100 ml after subpackaging in cryopreservation tubes. Well frozen storage tube box, marked on the box cover, placed in a -80 ℃ refrigerator for one-step frozen storage, frozen for 6 months, when the activation operation, take out the product from the refrigerator, immediately placed in a 35 ℃ water bath for rapid Shake to revive until the internal icing is completely dissolved, inoculate the mycelia in each cryotube in PDA medium respectively, inoculate 5 dishes in each cryotube, cultivate at 25°C, and observe the growth of Pleurotus ost...

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Abstract

The invention discloses a protective agent for preserving strains and a preservation method. The protective agent for preserving strains is prepared from the following raw materials in percentage by mass: 8-12% of glycerin, 3-8% of a trehalose solution, and the balance of deionized water. The specific preparation method of the protective agent for preserving strains includes the steps that the rawmaterials are prepared according to the mass percentage of the protective agent for preserving strains, and glycerin, the trehalose solution and deionized water are mixed and then sterilized at 120-122DEG C for 20min to obtain the protective agent for preserving strains. The uneven state of a stain sample suspended in the protective agent can be eliminated; the quality of products can be ensured;a good environment is created for the passage growth; the stability of protein is ensured; and during the cryopreservation process, implementing of the programmed freezing is not needed, and the search and activation operations are convenient.

Description

technical field [0001] The method belongs to the technical field of biological fermentation, and in particular relates to a preserved strain protective agent and a preservation method. Background technique [0002] Several preservation methods commonly used in industrial production include: regular transfer method, liquid paraffin preservation method, sand tube preservation method, freeze vacuum drying method, liquid nitrogen ultra-low temperature cryopreservation method, -80°C cryopreservation method, etc. Each of these preservation methods has its advantages and disadvantages. For example, in the regular transfer method, the water in the culture medium on the slant is easy to evaporate during storage and shrink, dry up, collapse, and change color, which increases its concentration and osmotic pressure, which will cause the strain to degenerate or even die; some favorable traits after continuous passage Will weaken; easy to cause bacterial contamination. Liquid paraffin o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04
CPCC12N1/04
Inventor 冯占李贺文邱丽雯余养朝
Owner 江苏华绿生物科技股份有限公司
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