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Preparation method for high-purity 7S glycinin and application of preparation method

A technology of soybean globin and protein, which is applied in the preparation method of peptides, chemical instruments and methods, plant peptides, etc., can solve the problems of limiting the application of two kinds of proteins, complicated and lengthy separation process, and low protein purity, and achieves easy assembly line separation Purification, simple process, and the effect of improving purity

Inactive Publication Date: 2014-04-23
北京龙科方舟生物工程技术有限公司
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, since the isoelectric points of 11S and 7S soybean globinin are not much different, this method often requires multi-step separation, and the precision of separation is improved by means of low-temperature equipment and high-speed centrifuge. The separation process is complicated and lengthy, and most of them can only be used in laboratories. Sample preparation; secondly, the purity of the proteins separated by most methods is low, which limits the application of the two proteins in actual production. At present, there is no effective method that is relatively simple and can be used for industrial production.

Method used

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  • Preparation method for high-purity 7S glycinin and application of preparation method
  • Preparation method for high-purity 7S glycinin and application of preparation method
  • Preparation method for high-purity 7S glycinin and application of preparation method

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Embodiment 1、7

[0055] The separation and purification of embodiment 1,7S glycinin

[0056] 1. Crude Extraction of 7S Glycinin

[0057] (1) Add defatted soybean powder into 50mM Tris-HCl buffer (containing 10mM mercaptoethanol) at a ratio of 1g:20ml, extract at 37°C for 1 hour, centrifuge at 10,000rpm at 28°C for 15 minutes, and collect the supernatant.

[0058] (2) Adjust the pH of the supernatant to 6.4 with 2 mol / L HCl, centrifuge at 10,000 rpm at 28°C for 15 minutes, and collect the supernatant again.

[0059] (3) Adjust the pH of the supernatant to 4.8 with 2 mol / L HCl, centrifuge at 10,000 rpm at 28°C for 15 minutes, and collect the precipitate.

[0060] (4) The precipitate was dissolved in 50mM Tris-HCl buffer solution with pH 8.0, and the filtrate was collected by filtration through a 0.45μm filter membrane.

[0061] 2. Strong anion exchange chromatography of 7S glycinin

[0062] (1) Take 10ml of Capto Q filler and fill it into an XK16 / 20 column (the inner diameter of the column is...

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Abstract

The invention discloses a preparation method for high-purity 7S glycinin and an application of the preparation method. The invention discloses the preparation method for the 7S glycinin. The preparation method comprises the following steps: (1) carrying out an acid precipitation method on defatted soybean powder to obtain a protein rough extracting solution; (2) carrying out strong anion exchange chromatography on the rough extracting solution of the step (1) and separating and purifying; collecting a target eluting solution 1; and (3) carrying out ceramic hydroxyapatite chromatography on the target eluting solution 1 and separating and purifying; and collecting a target eluting solution 2, namely a target protein. The target protein is the glycinin with the sedimentation coefficient of 7S. The method disclosed by the invention is simple process, is easy for assembly line separation and purification and is convenient for industrial production of the 7S glycinin.

Description

technical field [0001] The invention relates to a preparation method and application of high-purity 7S soybean globinin. Background technique [0002] Soybean is not only an excellent oil crop, but also an ideal edible protein resource. With the development of modern technology and the deepening of people's understanding of the nutritional and functional properties of soybean protein, defatted soybean flour with high protein content and its separated and purified protein have become Widely used in animal breeding, food processing and other fields. [0003] According to the sedimentation coefficient of ultracentrifugation, soybean protein can be divided into 2S, 7S, 11S and 15S glycinin. 7S and 11S globulins account for more than 80% of the total soybean protein content, and there are significant differences in their functional properties (water solubility, emulsification, surface activity, etc.) and nutritional properties. Egg whites are highly emulsifying. In recent year...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C07K1/36C07K1/30C07K1/18C07K1/14
CPCC07K14/415
Inventor 谯仕彦白晶宋青龙何涛
Owner 北京龙科方舟生物工程技术有限公司
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