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Preparation method of high-purity 11S glycinin and application thereof

A technology of soybean globin and protein, which is applied in the direction of chemical instruments and methods, plant peptides, peptide sources, etc., can solve the problems of complicated and lengthy separation process, limit the application of two kinds of proteins, and low protein purity, and achieve simple process and easy assembly line Separation and purification, the effect of improving purity

Inactive Publication Date: 2014-12-31
北京龙科方舟生物工程技术有限公司
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  • Abstract
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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 experiments. secondly, the purity of the proteins separated by most of the methods is low, which limits the application of the two proteins in actual production. At present, there is no effective method with a relatively simple process and can be oriented to industrial production.

Method used

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  • Preparation method of high-purity 11S glycinin and application thereof
  • Preparation method of high-purity 11S glycinin and application thereof
  • Preparation method of high-purity 11S glycinin and application thereof

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Embodiment 1、11

[0055] The separation and purification of embodiment 1, 11S glycinin

[0056] 1. Crude Extraction of 11S Glycinin

[0057] (1) Mix defatted soybean powder with 50mM Tris-HCl buffer (containing 10mM mercaptoethanol) at a volume 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 for 15 minutes at 28°C, and collect the precipitate.

[0059] (3) Dissolve the precipitate with 50 mM Tris-HCl buffer solution with pH 8.0, and collect the filtrate by filtering through a 0.45 μm filter membrane.

[0060] 2. Strong anion exchange chromatography of 11S glycinin

[0061] (1) Take 10ml of Capto Q filler and fill it into an XK16 / 20 column (inner diameter 1.6cm, filler height 5cm). After filling, equilibrate 10 column volumes with 50mM Tris-HCl buffer, pH 8.0.

[0062] (2) Put the filtrate obtained in step 1 on the column,...

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Abstract

The invention discloses a preparation method of high-purity 11S glycinin and an application thereof. The preparation method of high-purity 11S glycinin, disclosed by the invention, comprises the following steps: (1), obtaining a protein crude extraction liquid from defatted soybean powder through an acid precipitation method; (2), performing strong anion exchange chromatography on the crude extraction solution in the step (1) for separation and purification, and collecting a target eluant 1; and (3), performing ceramic hydroxyapatite chromatography on the target eluant 1 for separation and purification, and collecting a target eluant 2, namely, a target protein, wherein the target protein is glycinin with a sedimentation coefficient 11S. The preparation method disclosed by the invention is simple in process, easy to separate and purify in a production line, and convenient to produce industrially.

Description

technical field [0001] The invention relates to a preparation method and application of high-purity 11S 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. [0004] The ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C07K1/36C07K1/30C07K1/34
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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