A method for combined separation and purification of protein by reverse micelle extraction and pulsed electric field

A pulsed electric field and reverse micelle technology, applied in the field of reverse micelle extraction-pulsed electric field combined separation and purification of proteins, can solve the problems of lack of driving force and difficulty in entering micelles for large protein molecules, achieving low cost, high extraction rate, Avoid shear damage effects

Inactive Publication Date: 2020-08-18
TAIZHOU UNIV
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Problems solved by technology

The main reason is that although the mixed reverse micelles system can form a large water pool to accommodate macromolecular proteins, it is difficult for large protein molecules to enter the micelles and lack the driving force to enter the interior of the micelles.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Extraction of rapeseed protein from rapeseed meal by pulsed electric field-assisted reverse micellar method

[0036] Rapeseed meal is a by-product of rapeseed oil extraction process, which contains 35%-45% high-quality vegetable protein. The rapeseed protein liquid extracted from rapeseed meal can be widely used in food additives, functional foods and other related fields, and has broad application prospects. The current extraction methods mainly include aqueous phase method, aqueous phase enzymatic hydrolysis method, acetone extraction method, two-liquid phase extraction method, ultrafiltration and diafiltration methods, etc. These traditional methods all have problems such as long extraction time, low extraction efficiency or large amount of organic solvent used. It has been proved that rapeseed protein can be extracted from rapeseed meal by pulsed electric field-assisted reverse micellar method. The main process is as follows:

[0037] (1) Pretreatment o...

Embodiment 2

[0044] Example 2 Pulsed electric field assisted reverse micellar method to extract soybean protein from soybean raw materials

[0045] The plant protein content in soybean is rich, up to about 40%, and has high nutritional value, close to complete protein, and is an important raw material of plant protein. At present, the industrial extraction method of soybean protein is mainly the traditional alkali-soluble acid precipitation method. This method introduces a large amount of acid and alkali in the production process, which not only has high production costs, but also easily causes a series of environmental pollution problems. lead to changes in protein fold configuration. At present, some new processes have emerged, such as the use of reverse micelles to extract soybean protein, but the extraction time and efficiency need to be further improved. The invention utilizes the method of pulsed electric field assisted reverse micelles, and proves that the method can be used to rea...

Embodiment 3

[0053] Example 3 Extraction of phycocyanin from Spirulina Platensis by pulsed electric field-assisted reverse micellar method

[0054] Spirulina platensis is rich in protein, vitamins and fatty acids, among which the protein content is as high as 50%-70% of the dry weight of the cells. Among the proteins contained, phycocyanin has strong biological activity, and has the functions of anti-radiation, antibacterial and anti-inflammatory, anti-oxidation, anti-tumor, enhancing immunity, assisting hematopoietic stem cell repair, and protecting human nerve tissue from damage. About 7% of the total protein. Our country produces 900 tons of spirulina powder per year, but the processing capacity of the deep-processed product cyanobacteria protein is relatively weak. At present, it is mainly purified by multi-stage column chromatography. The steps of extraction and purification are long and the cost is high. The invention provides a method for extracting phycocyanin from Spirulina plate...

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Abstract

The invention discloses a method for separating and purifying protein by combining reverse micelle extraction with a pulsed electric field. The method comprises the following steps: (1) adding a target protein raw material into a reverse micelle solution, evenly mixing and then transferring to an extraction cavity of the pulsed electric field, wherein the extraction cavity of the pulsed electric field is arranged in a cooling tank, and implementing pulsed electric field combined with reverse micelle extraction; (2) carrying out solid-liquid separation on the solution, treated in the step (1),in the extraction cavity of the pulsed electric field, taking supernatant to obtain a pre-extracted solution of the target protein; (3) carrying out back extraction on the obtained pre-extracted solution to obtain post-extraction liquid containing the target protein; (4) concentrating and drying the post-extraction liquid to obtain the target protein. The method can increase the extraction yield of the protein and shorten the extraction time; furthermore, the quality of the product is also obviously superior to that of products obtained by using the traditional extraction method.

Description

technical field [0001] The invention relates to the technical field of protein separation and purification, in particular to a method for separating and purifying proteins combined with reverse micellar extraction and pulsed electric field. Background technique [0002] With the development of genetic engineering and cell engineering, traditional solvent extraction technology is widely used in the production of antibiotics and other substances, and shows excellent separation performance, but it is difficult to apply to the extraction and separation of proteins. One is that most proteins are insoluble in organic solvents and are easily deformed when exposed to organic solvents. The other is that the surface of protein molecules has a lot of charges, and it is difficult for ordinary ion-associated extractants to work. In recent years, with the development of reverse micellar extraction technology, a new technology with good development prospects has been provided for protein s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/36C07K1/24C07K1/14
CPCC07K1/14C07K1/24C07K1/36
Inventor 章志成袁玲姚俊孔庆娜
Owner TAIZHOU UNIV
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