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
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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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