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

Inactive Publication Date: 2015-04-22
合肥巅峰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the deficiencies in the prior art, and provide buffer and application thereof for normal temperature preservation of recombinant Beauveria bassiana proteinase K, to solve the problem that recombinant Beauveria bassiana proteinase K cannot be preserved at normal temperature, and the storage time short technical question

Method used

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  • Buffer solution for normal-temperature preservation of recombinant beauveria brongniartii protease K and applications thereof
  • Buffer solution for normal-temperature preservation of recombinant beauveria brongniartii protease K and applications thereof
  • Buffer solution for normal-temperature preservation of recombinant beauveria brongniartii protease K and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a buffer solution for the normal-temperature preservation of recombinant beauveria brongniartii protease K and applications thereof. The buffer solution comprises the following components: trihydroxymethyl aminomethane Tris, a soluble sodium salt, a soluble calcium salt, glycerin, and the balance of water, wherein the pH value of the buffer solution is 7.5-8.4. Compared with the prior art, the buffer solution can significantly enhance the time of normal-temperature preservation of recombinant beauveria brongniartii protease K and does not degrade the catalytic efficiency of the recombinant beauveria brongniartii protease K; and the recombinant beauveria brongniartii protease K stored by using the buffer solution disclosed by the invention is directly applied without being processed, such as being applied to the extraction of nucleosides, and the like, so that the transportation, storage and application costs of the recombinant beauveria brongniartii protease K are significantly reduced, therefore, the buffer solution has a wide application prospect.

Description

technical field [0001] The invention relates to a protease K buffer, in particular to a buffer for normal temperature preservation of recombinant Beauveria bassiana protease K and its application. Background technique [0002] Proteinase K (Proteinase K), also known as endopeptidase K, saprophytic fungal alkaline protease, Candida albicans serine protease, Candida albicans protease K, is a serine protease with broad cutting activity. The protein degrades native keratin by cleaving the carboxy-terminal peptide bonds of aliphatic and aromatic amino acids, hence its name. Because proteinase K can exist stably in guanidine hydrochloride (3M), urea (4M) and SDS (0.5-1%) and has the ability to degrade proteins, it is widely used in the preparation of chromosomal DNA for pulse electrophoresis, Western blot and removal Nucleases in DNA and RNA preparations. The general working concentration of proteinase K is 50-100 μg / ml, and it is active in a wide pH range (pH4-12). [0003] Pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/96C12N9/58
CPCC12N9/96C12N9/6472
Inventor 李旭滕脉坤
Owner 合肥巅峰生物科技有限公司
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