Alternate current-aided efficient protein enzymolysis method

An auxiliary protein and auxiliary enzymatic hydrolysis technology, applied in the field of proteomics, can solve problems such as restricting the speed of protein analysis, achieve good application prospects, improve work efficiency, and save enzymatic hydrolysis time

Inactive Publication Date: 2008-10-08
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the traditional solution enzymatic hydrolysis method is time-consuming (more than 12 hours), it directly restricts the improvem...

Method used

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  • Alternate current-aided efficient protein enzymolysis method
  • Alternate current-aided efficient protein enzymolysis method
  • Alternate current-aided efficient protein enzymolysis method

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Embodiment Construction

[0026] Further describe the present invention below by embodiment and accompanying drawing:

[0027] 1. In-tube alternating current assists rapid enzymatic hydrolysis of proteins

[0028] The structure of the in-pipe alternating current assisted protein enzymolysis device proposed by the present invention is as follows: figure 1 As shown, it is composed of a 0.5mL polypropylene centrifuge tube 4, a straight platinum wire electrode 1, a spiral platinum wire electrode 2 and a cover 3. The alternating current applied to the electrode is directly reduced by a small transformer to a civilian 220V alternating current (50Hz). Pressed, the secondary coil of the transformer has a center tap, which can provide a low-voltage alternating current (50Hz) of 2-10V.

[0029] The protein was dissolved in 50 mmol / L ammonium bicarbonate solution (pH 8.0), and heated and denatured in a water bath at 95°C for 10 minutes to fully expose the enzyme cleavage site of the protein. Then mix with a cer...

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Abstract

The invention belongs to the proteomics technology field, in particular relates to an efficient enzymolysis method of alternating current accessory protein. Protein and proteolytic enzymes such, as trypsin, are solubilized in buffer solution in a certain proportion by the method, and the solution is ensured to contact with an electrode applied by alternating voltage; the charged protein, enzyme and inorganic ions, etc. change the direction of motion in the electric field immediately, and the motions, such as vibration, collision, rotation and friction, etc., of the molecules such as protein and enzyme, etc., are caused. Simultaneously, shakings also occurs to the charged group in the peptide chain of the protein molecules under the exertion of the alternating electric field, and more restriction sites are exposed. The motions improve the energy level and interaction frequency of the protein and enzyme molecules, in order to increase the protein enzymolysis efficiency greatly. By adopting the alternating current accessory protein enzymolysis, the enzymolysis time is greatly shortened from twelve hours of traditional solution enzymolysis to five to ten minutes, and the enzymolysis efficiency is improved remarkably.

Description

technical field [0001] The invention belongs to the technical field of proteomics, and in particular relates to an alternating current assisted protein high-efficiency enzymatic hydrolysis method. Background technique [0002] Proteomics is currently a research hotspot in the international life science community. It explores and discovers the laws of life activities and the essence of important physiological and pathological phenomena at the overall protein level and at a level that is closer to the essence of life. Among them, the identification of proteins is one of the most important tasks. Peptide mass fingerprint (Peptide mass fringprint, PMF) is a common method for studying protein structure information and identifying proteins, and enzymatic hydrolysis is a key step in protein analysis in proteomics research, the purpose is to decompose proteins into peptides, The peptide mixture was then analyzed by matrix-assisted laser desorption ionization time-of-flight mass spe...

Claims

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Application Information

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IPC IPC(8): C12P21/06
Inventor 陈刚王胜刘婷杨芃原
Owner FUDAN UNIV
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