Buffer solution for normal-temperature preservation of recombinant beauveria brongniartii protease K and applications thereof
A technology for storage of Beauveria bassiana at room temperature, which is applied in the fields of enzymes, enzyme stability, and hydrolytic enzymes. It can solve the problems of short storage time and the inability of recombinant Beauveria bassiana proteinase K to be stored at room temperature, so as to reduce transportation and enhance The time of storage at room temperature and the effect of wide application prospects
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Embodiment 1
[0033] The preparation method of the buffer that is used for the normal temperature preservation of recombinant Beauveria bassiana proteinase K of the present embodiment comprises the following steps:
[0034] (1) Accurately weigh 0.303g Tris-base and 0.147g calcium chloride dihydrate CaCl 2 and 1.17g of sodium chloride NaCl, then add 40ml of pure water, mix well, adjust its pH to 7.8 with hydrochloric acid, and set the volume to 50ml with a graduated cylinder to obtain a mixed solution.
[0035] (2) Accurately measure 50 mL of glycerin, add it into the above mixture and mix well to obtain the buffer.
[0036] The above-mentioned buffer solution is used for normal temperature preservation of recombinant Beauveria bassiana proteinase K, and its protective effect is determined, as follows:
[0037] A. BSA digestion test
[0038] a. Pretreatment of reaction substrate BSA
[0039] Dissolve 5mg of bovine serum albumin BSA in 6mol / L guanidine hydrochloride, 50mmol / L Tris-Cl (pH8....
Embodiment 2
[0082] The buffer solution provided in this example for the normal temperature preservation of recombinant Beauveria bassiana proteinase K comprises the following steps:
[0083] (1) Tris-base with a molar concentration of 10mmol / L, NaCl with a molar concentration of 20mmol / L, and CaSO with a molar concentration of 2mmol / L 4 Add water to dissolve, and adjust the pH value to 7.5 with HCl to obtain a mixed solution;
[0084] (2) Glycerol is added into the mixed solution in step (1) according to the volume percentage of 60% and mixed evenly, which is the buffer solution.
[0085] Dissolve the recombinant Beauveria bassiana protease K in the above buffer solution at a concentration of 10 mg / mL, and store it at room temperature for more than one year. The Beauveria bassiana protease K dissolved in the buffer solution K can be directly applied to fields such as nucleic acid extraction.
Embodiment 3
[0087] The buffer solution provided in this example for the normal temperature preservation of recombinant Beauveria bassiana proteinase K comprises the following steps:
[0088] (1) Tris-base with a molar concentration of 500mmol / L, NaCl with a molar concentration of 500mmol / L, and CaSO with a molar concentration of 50mmol / L 4 Add water to dissolve, and adjust the pH value to 8.4 with HCl to obtain a mixed solution;
[0089] (2) Glycerol is added to the mixture in step (1) at a volume percentage of 30% and mixed evenly, which is the buffer solution.
[0090] Dissolve the recombinant Beauveria bassiana protease K in the above-mentioned buffer at a concentration of 2 mg / mL, and then store it at room temperature for more than one year. The Beauveria bassiana protease dissolved in the buffer K can be directly applied to fields such as nucleic acid extraction.
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