Osmotic pressure stabilizer and use thereof

A technology of osmotic pressure and humectant, applied in the direction of using electricity/wave energy to treat microorganisms, fungi, microorganisms, etc. Mutation effect, high mutation efficiency, good survival effect

Active Publication Date: 2017-05-31
FOSHAN HAITIAN FLAVOURING & FOOD CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the prolongation of the action time, the volatilization of water in the trace system will have a great impact on the stability of the protoplasts, and the osmotic pressure will increase. The protoplasts after losing protection may cause irreparable shrinkage and dehydration.

Method used

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  • Osmotic pressure stabilizer and use thereof
  • Osmotic pressure stabilizer and use thereof
  • Osmotic pressure stabilizer and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] The preparation of embodiment 1 composition

[0070] Preparation of composition 1:

[0071] Formula: sodium chloride 0.8wt%,

[0072] Glycerin 5wt%,

[0073] Trehalose 20wt%,

[0074] The balance is Tris-HCl buffer.

[0075] Sodium chloride, glycerin, and trehalose in the above formula were dissolved in Tris-HCl buffer, mixed evenly, and the pH value was adjusted to 7.10 to obtain composition 1.

[0076] Preparation of Tris-HCl buffer solution used in Composition 1: Mix 50ml of 0.1mol / L tris-hydroxymethylaminomethane (Tris) solution with 45.7ml of 0.1mol / L hydrochloric acid, and then dilute with water to 100ml.

[0077] Preparation of composition 2:

[0078] Formula: sodium chloride 0.8wt%,

[0079] Hyaluronic Acid 5wt%,

[0080] Glycerin 5wt%,

[0081] Polyethylene glycol 5wt%,

[0082] The balance is Tris-HCl buffer.

[0083] Sodium chloride, hyaluronic acid, glycerin, and polyethylene glycol in the above formula were dissolved in Tris-HCl buffer, mixed even...

Embodiment 2

[0093] The establishment of embodiment 2 ARTP mutagenesis method

[0094] The establishment process of the ARTP mutagenesis method is as follows: figure 1 shown.

[0095] 1) Obtain fungal protozoa according to conventional methods

[0096] Protoplasts are prepared from newly germinated fungal spores (such as Aspergillus oryzae, Trichoderma viride, Pleurotus eryngii, Aspergillus niger, etc.). Prepare a concentration of 5×10 8 The single spore suspension with a concentration of CFU / ml is inoculated into the liquid medium according to the inoculation amount of 2%. Under the condition of the shaker speed of 120rpm / min and the set temperature of 32°C, shake the flask for 15 hours until most of the spores are just During germination, the G3 sand core funnel filters out the remaining spores, and the spores and mycelia that have just germinated are collected by centrifugation. An appropriate amount of compound enzyme solution was added for wall breaking treatment for 3.5 hours, an...

Embodiment 3-5

[0111] The selection of protoplast preparation method and ARTP treatment time is basically the same as that of Example 2.

[0112] Contrast method 1 is protoplast ultraviolet mutagenesis method, after this method prepares fungal protoplast earlier, adjust protoplast concentration with conventional osmotic pressure stabilizer (sodium chloride solution of 0.8mol / L) be 3 * 10 6 CFU / mL-3×10 8 CFU / mL to obtain the protoplast sample to be mutated. Take 5ml of the protoplast sample to be mutated, spread it on a conventional petri dish, and carry out ultraviolet mutagenesis treatment. The UV lamp was preheated for 15 minutes, the power of the UV lamp was 10W, the vertical distance from the irradiation position was 15cm, and the UV treatment time was uniformly set to 20 seconds.

[0113] The control method 2 is the ARTP fungal mutagenesis method, in which the fungal spores are directly mutagenized with ATTP, and the preparation of the mutagenized sample is the same as that described ...

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Abstract

The invention relates to a composition, which contains an osmotic pressure maintenance agent, a humectant and a pH buffer, wherein the pH value of the composition is 6.5 to 7.5. As an osmotic pressure stabilizer, the composition has a protection effect on a protoplast, and the protoplast can be prevented from being deactivated. When the osmotic pressure stabilizer composition is adopted, ARTP (atmospheric room temperature plasma) mutagenesis is performed on the protoplast, so that the mutation efficiency can be improved. The invention also relates to a method for mutagenizing the protoplast by virtue of the composition as the osmotic pressure stabilizer.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a composition and its application. As an osmotic pressure stabilizer, the composition has a protective effect on protoplasts and can prevent the protoplasts from inactivating. The invention also relates to a method for using the composition as an osmotic pressure stabilizer to mutate protoplasts. Background technique [0002] Protoplast mutagenesis is a process in which protoplasts are directly treated by mutagenesis, and then propagated and regenerated to obtain mutants. Because protoplasts do not have a cell wall barrier, mutagens can easily enter the cell, and the probability of mutation is greater, the time is shorter, and the efficiency is higher. [0003] Atmospheric Room Temperature Plasma (ARTP) mutagenesis is a new and efficient physical mutagenesis method. ARTP mutagenesis technology adopts atmospheric pressure radio frequency glow discharge plasma source to excite plasma ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N13/00
CPCC12N1/14C12N13/00
Inventor 黄艳童星
Owner FOSHAN HAITIAN FLAVOURING & FOOD CO LTD
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