Preparation method of kidney bean agglutinin with low blood coagulation activity and high and alpha-glucosidase activity

A technology of glucosidase and hemagglutination activity, which is applied to the preparation methods of peptides, protease inhibitors, chemical instruments and methods, etc. The effect of reducing the hemagglutination activity of kidney bean lectin and simple operation

Active Publication Date: 2013-05-08
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention provides a novel preparation and processing method for preparing kidney bean agglutinin with low hemagglutination activity and high α-glucosidase inhibitory activity, in order to solv

Method used

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  • Preparation method of kidney bean agglutinin with low blood coagulation activity and high and alpha-glucosidase activity

Examples

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Embodiment 1

[0029] (1) Preparation of kidney bean crude extract: mix kidney beans with Tris buffer and homogenate, the ratio of solid to liquid is 1:15, let stand at 4°C for 24h, filter with four layers of gauze, centrifuge (6000g, 20min), and take the supernatant , to obtain kidney bean extract;

[0030] (2) Ammonium sulfate precipitation: add solid (NH 4 ) 2 SO 4 To 60% saturation, let stand at 4°C overnight, centrifuge (5000g, 30min) to get the precipitate. The precipitate was dissolved in distilled water, dialyzed at 4°C for 12 hours, desalted, and freeze-dried to obtain the crude kidney bean lectin;

[0031] (3) Affinity chromatography: use Affi-gel blue column (20cm*2.5cm), mobile phase A is 20 mmol / L tris-HCl, pH 7.4, flow rate is 1mL / min, and use mobile phase A to dissolve the previous step of separation The fractions obtained were purified. Mobile phase B is to add NaCl to mobile phase A to 1.2 mol / L, and use mobile phase B to carry out elution. Use 280 nm ultraviolet l...

Embodiment 2

[0037] (1) Preparation of crude kidney bean extract: crush kidney beans and mix with Tris buffer for 8 hours, the ratio of solid to liquid is 1:15, let stand at 4°C for 16 hours, filter with four layers of gauze and centrifuge (6000g, 20min) to obtain crude kidney beans Extract;

[0038] (2) Ammonium sulfate precipitation: add solid (NH 4 ) 2 SO 4 To 60% saturation, let it stand overnight at 4°C, and centrifuge (5000g, 30min) to collect the precipitate. The precipitate was dissolved in distilled water, dialyzed at 4°C for 12 hours, desalted, and freeze-dried to obtain the crude kidney bean lectin;

[0039] (3) Affinity chromatography: use Affi-gel blue column (20cm*2.5cm), mobile phase A is 50 mmol / L tris-HCl, pH 7.2, flow rate is 1mL / min, and use mobile phase A to dissolve the previous step of separation The fractions obtained were purified. Mobile phase B is to add NaCl to 1.4 mol / L in mobile phase A, and use it for elution. Use 280 nm ultraviolet light to detect, coll...

Embodiment 3

[0045] (1) Preparation of kidney bean extract: Mix kidney beans and Tris buffer for homogenization, the ratio of solid to liquid is 1:15, let stand at 4°C for 24 hours, filter with four layers of gauze and centrifuge (8000g, 20min), take the supernatant, Kidney bean extract;

[0046] (2) Ammonium sulfate precipitation: add solid (NH 4 ) 2 SO 4 To 65% saturation, let stand overnight at 4°C, and centrifuge (5000g, 30min) to collect the precipitate. The precipitate was dissolved in distilled water, dialyzed at 4°C for 12 hours, desalted, and freeze-dried to obtain the crude kidney bean lectin;

[0047] (3) Affinity chromatography: use Affi-gel blue column (20cm*2.5cm), mobile phase A is 20 mmol / L tris-HCl, pH 7.4, flow rate is 1mL / min, and use mobile phase A to dissolve the previous step of separation The fractions obtained were purified. Mobile phase B was added NaCl to mobile phase A to 1.2 mol / L, and mobile phase B was used for elution. Use 280 nm ultraviolet light to de...

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Abstract

The invention relates to a preparation method of a kidney bean agglutinin with low blood coagulation activity and high and alpha-glucosidase activity. The preparation method disclosed by the invention specifically comprises the following steps: preparing a kidney bean crude extract; precipitating via ammonium sulfate; carrying out affinity chromatography; carrying out ion-exchange chromatography; homogenizing at a pressure of 10-50 MPa; treating via a pulse electric field at an electric field strength of 5-50 kV/cm; and taking the treated kidney bean agglutinin, freezing and drying to obtain kidney bean agglutinin powder. According to the discovery of the invention, after the kidney bean agglutinin is treated by the method, compared with the thermal treatment carried out for 20-40 minutes at the temperature of 80-120 DEG C, the blood coagulation activity of the kidney bean agglutinin is reduced and most of alpha-glucosidase inhibitory activity is kept, namely, on the one hand, the nutrition resistance of the kidney beans is greatly reduced and on the other hand, the beneficial alpha-glucosidase inhibitory activity is improved.

Description

technical field [0001] The invention relates to the field of food biochemistry, in particular to a novel preparation method capable of effectively reducing the hemagglutination activity of kidney bean agglutinin while maintaining its high alpha-glucosidase inhibitory activity. Background technique [0002] Kidney bean lectin ( Phytohemagglutinin, PHA ) is a kind of glycoprotein of non-immune origin in kidney bean grains. It has been reported in the literature that it has dual characteristics: on the one hand, because lectin is one of the components formed during the long-term evolution of plants to resist diseases and insect pests and animal digestion, it is harmful to the human body. It has a certain anti-nutritional effect, can reversibly bind specific monosaccharides or oligosaccharides, and promote agglutination or precipitation of red blood cells; on the other hand, it also has α-glucosidase inhibitory activity, anti-HIV virus and inhibition of T lymphocytes Benefic...

Claims

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Application Information

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IPC IPC(8): C07K14/81C07K1/36C07K1/30C07K1/22C07K1/18
Inventor 任娇艳赵谋明崔春刘岑岑
Owner SOUTH CHINA UNIV OF TECH
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