Method for microwave-assisted reverse micelle separation and purification of kidney bean agglutinin

A separation and purification, microwave-assisted technology, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve the problems of low separation and purification ratio, multiple processes, and high cost of kidney bean lectin, and achieve easy continuous production and Effects of industrial scale-up, reduction of environmental pollution, and low equipment requirements

Inactive Publication Date: 2013-05-01
HARBIN INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to overcome the deficiencies of the prior art, solve the problems of low separation and purification ratio of kidney bean lectin, high cost, many processes and long time, and provide a microwave-assisted reverse glue with high efficiency, easy operation, low cost, safety and environmental protection Method for beam separation and purification of kidney bean lectin

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  • Method for microwave-assisted reverse micelle separation and purification of kidney bean agglutinin
  • Method for microwave-assisted reverse micelle separation and purification of kidney bean agglutinin

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Embodiment 1

[0042] A method for separating and purifying kidney bean lectin by anionic surfactant microwave-assisted reverse micelles of the present invention specifically comprises the following steps:

[0043] (1) Preparation of kidney bean lectin aqueous extract:

[0044] Select high-quality kidney bean powder, pass through a 80-mesh sieve, dissolve in 10 times (volume) phosphate buffer solution (pH7.2, 25mM), microwave-assisted extraction at 25°C for 30min (325W), centrifuge at 12000rpm, 4°C for 20min, and take supernatant.

[0045] (2) Preparation of reverse micellar solution:

[0046] Dissolve the anionic surfactant sodium succinate-2-ethylhexylsulfonate (AOT) completely in isooctane to prepare a reverse micellar solution with a concentration of 130mM. Add a certain amount of water to the reverse micellar system W o The value is 25. In the present embodiment, the cosolvent is butanol, and the volume ratio of the cosolvent addition is 10%.

[0047] (3) Pre-extraction process:

...

Embodiment 2

[0053] A cationic surfactant microwave-assisted reverse micellar separation and purification method for kidney bean lectin of the present invention specifically comprises the following steps:

[0054] (1) Preparation of kidney bean lectin aqueous extract:

[0055] Select high-quality kidney bean powder, pass through a 80-mesh sieve, dissolve in 10 times (volume) phosphate buffer solution (pH7.2, 25mM), microwave-assisted extraction at 25°C for 30min (325W), centrifuge at 12000rpm, 4°C for 20min, and take supernatant.

[0056] (2) Preparation of reverse micellar solution:

[0057] The cationic surfactant cetyltrimethylamine bromide / cetyltrimethylammonium bromide (CTAB) was completely dissolved in isooctane to prepare a reverse micellar solution with a concentration of 150 mM. Add a certain amount of water to the reverse micellar system W o The value is 25. In the present embodiment, the cosolvent is butanol, and the volume ratio of the cosolvent addition is 10%.

[0058] (...

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Abstract

The invention discloses a method for microwave-assisted reverse micelle separation and purification of kidney bean agglutinin, and relates to a separation and purification method of the agglutinin, aiming at solving the problems of low ratio, high cost, more processes and long time for separation and purification of the kidney bean agglutinin. According to the method for the microwave-assisted reverse micelle separation and purification of the kidney bean agglutinin, the agglutinin is directly separated and purified from kidney bean agglutinin water extract; and the method comprises a pre-extraction process and a back extraction process. Under an auxiliary effect of microwave, water-soluble protein in kidney bean is quickly dissolved into the water extract, so that the dissolubility is greatly improved, and the leaching time is shortened. Based on a liquid-liquid extraction principle, the selectivity of extraction is improved, and high-efficiency pre-extraction and back extraction of the kidney bean agglutinin can be realized. By applying a reverse micelle extraction technology, a better separation and purification effect can be achieved with only the pre-extraction and back extraction steps; the method has simple steps and is convenient to operate; and the pre-treatment time can be greatly shortened in combination with microwave assistance so as to save the cost.

Description

technical field [0001] The invention relates to a method for separating and purifying lectin, in particular to a method for directly separating and purifying kidney bean lectin from water extract by using reverse micellar extraction technology. Background technique [0002] Lectin (lectin or agglutinin) is a kind of non-immune glycoprotein or sugar that has sugar specificity and can bind to the oligosaccharide structure of special glycoproteins and glycolipids on the cell surface to promote cell agglutination or sugar complex precipitation. binding protein. Lectins have a wide range of uses. In biochemical research, they can be used for the separation and purification of sugar-containing polymers and the identification of sugar chain structures; in biological research, they can specifically identify cell surface signal molecules, and can be used to select mutant cell lines and study cellular mechanisms. and cell surface characteristics and identification of microorganisms; ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/42C07K1/14
Inventor 马莺何述栋李琳何胜华王超
Owner HARBIN INST OF TECH
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