Method for separating and purifying soybean agglutinin through reverse micelle

A soybean lectin, separation and purification technology, which is applied in the field of chemical separation, can solve the problems of low separation purity, high cost, and many processes of soybean lectin, and achieves easy continuous production and industrial amplification, low equipment requirements, and reduced environmental pollution. Effect

Active Publication Date: 2016-12-07
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art, solve the problems of low purity, high cost, multiple processes, and long time for the separation of soybean lectin, and provide a high-efficiency, easy-to-operate, low-cost, safe and environmentally friendly reverse micellar separation and purification Soy Lectin Methods

Method used

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  • Method for separating and purifying soybean agglutinin through reverse micelle
  • Method for separating and purifying soybean agglutinin through reverse micelle

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Effect test

Embodiment 1

[0030] (1) Surfactant preparation process:

[0031] Take 50g of pine bark extract, dissolve it in 50g of acetone, add 35g of pyridine as an ancidic agent at -10°C, then slowly add 50g of hexanoyl chloride dropwise, react for 1 hour, add water to dilute the reaction solution. Separate the organic phase, extract the aqueous phase with 50g ethyl acetate, combine the organic phases to obtain the esterified grafted pine bark extract solution; use 1 times the volume of 1mol / L HCl, saturated Na 2 CO 3 solution, saturated NaCl solution, and vacuum-dried to obtain a surfactant.

[0032] (2) Preparation of soybean lectin water extract: Take 50g of high-quality soybean powder, pass through a 40-mesh sieve, dissolve in 250mL of distilled water, and extract with ultrasonic assistance for 60min (120W) at 10°C. After extraction, centrifuge at 12000rpm and 4°C 15min, take the supernatant;

[0033] (3) Preparation of reverse micelle solution: 45g surfactant and 2g co-solvent glycerin are co...

Embodiment 2

[0038] (1) Surfactant preparation process:

[0039]Take 50g of pine bark extract, dissolve it in 200g of acetone, add 50g of pyridine as an ancidic agent at -10°C, then slowly add 60g of octanoyl chloride dropwise, react for 2 hours, and then add water to dilute the reaction solution. Separate the organic phase, extract the aqueous phase with 50g ethyl acetate, combine the organic phases to obtain the esterified grafted pine bark extract solution; use 1.5 times the volume of 1mol / L HCl, saturated Na 2 CO 3 solution, saturated NaCl solution, and vacuum-dried to obtain a surfactant.

[0040] (2) Preparation of soybean lectin water extract: Take 50g of high-quality green soybeans, grind them through a 70-mesh sieve, dissolve them in 500mL of distilled water, and extract them with ultrasonic assistance for 30min (120W) at 25°C. After extraction, centrifuge at 12000rpm and 4°C 15min, take the supernatant;

[0041] (3) Preparation of reverse micellar solution: 70g surfactant and ...

Embodiment 3

[0046] (1) Surfactant preparation process:

[0047] Take 50g of pine bark extract, dissolve it in 150g of acetone, add 60g of pyridine as an acid application agent at -10°C, then slowly add 95g of myristoyl chloride dropwise, react for 8 hours, add water to dilute the reaction solution. Separate the organic phase, extract the aqueous phase with 12.5g ethyl acetate, combine the organic phases to obtain the esterified grafted pine bark extract solution; use 4 times the volume of 1mol / L HCl, saturated Na 2 CO 3 solution, saturated NaCl solution, and vacuum-dried to obtain a surfactant.

[0048] (2) Preparation of soybean lectin water extract: high-quality black soybean powder, passed through a 100-mesh sieve, dissolved in 50 times (volume) distilled water, ultrasonically assisted extraction at 60°C for 5min (120W), after extraction, it was subjected to 12000rpm, 4 Centrifuge at ℃ for 15 min, and take the supernatant;

[0049] (3) Preparation of reverse micellar solution: 130g ...

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Abstract

The invention provides a method for separating and purifying soybean agglutinin through reverse micelle. The method includes the steps of preparation of soybean agglutinin water extract, preparation of a reverse micelle solution, a forward extraction process and a reverse extraction process. The method for separating and purifying soybean agglutinin is short in needed time, low in requirement on equipment, low in implementation cost and rapid and easy and convenient to operate, continuous production and industrial enlargement are easy, separation, purification and concentration can be completed at the same time, the purity of soybean agglutinin is 85% or above, the yield of soybean agglutinin is 85% or above, structure modified pine bark extract is adopted as a surfactant in the extraction process, soybean agglutinin can be effectively protected and is not prone to be inactivated, purity is high, and a high-purity product can be obtained in the following process only through one step of simple ion exchange chromatography or chromatography.

Description

technical field [0001] The invention belongs to the technical field of chemical separation, and in particular relates to a method for separating and purifying soybean lectin, in particular to a method for separating and purifying soybean lectin by reverse micelles. 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 aggregation 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/42C07K1/18C07K1/14
CPCC07K14/42
Inventor 黄洁周良普闫晓玉童耀阳金志敏
Owner ZHEJIANG UNIV OF TECH
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