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Method for separating wheat alcohol-soluble protein by utilizing sucrose density gradient centrifugation method

A wheat gliadin and sucrose density gradient technology is used in the separation and purification of wheat gluten protein, and the sucrose density gradient centrifugation method is used to separate or purify wheat gliadin. Protein separation and purification issues

Inactive Publication Date: 2018-02-02
HENAN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, these classification methods are not very perfect and detailed, and there are still problems that cannot completely cover protein separation and purification in specific production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0006] Take 5 mL of 70% ethanol-extracted wheat prolamin solution (solution to be separated), inject it into the bottom of a 50 mL centrifuge tube, add 10 mL of prepared 20%, 40% and 60% sucrose solutions, and follow the layer laying method Inject from the bottom of the centrifuge tube, and finally from the bottom up: 60%-40%-20%-prolamin solution to be separated, in a horizontal centrifuge, after centrifuging at 5000 g for 30 min, respectively from 20% sucrose solution, 40% % sucrose solution and 60% sucrose solution to absorb a certain amount of solution, and use a micro-ultraviolet-visible spectrophotometer to detect the protein content in the solution at 280 nm, and the protein content in the upper layer (20% sucrose solution) is 6.2 mg / mL, The middle (40% sucrose solution) had a protein content of 5.4 mg / mL and the lower layer (60% sucrose solution) had a protein content of 2.1 mg / mL. In this way, the sucrose solutions in the upper layer, middle layer and lower layer are ...

Embodiment 2

[0008] Take 3 mL of 70% ethanol-extracted wheat prolamin solution (solution to be separated), inject it into the bottom of a 50 mL centrifuge tube, add 10 mL of prepared 10%, 30% and 50% sucrose solutions, and follow the layer laying method Inject from the bottom of the centrifuge tube, and finally from the bottom up: 50%-30%-10%-prolamin solution to be separated, in a horizontal centrifuge, after centrifuging at 10 000 g for 30 min, respectively from 10% sucrose solution, A certain amount of solution was drawn from 30% sucrose solution and 50% sucrose solution, and the protein content in the solution was detected by a micro-ultraviolet-visible spectrophotometer at 280 nm, and the protein content in the upper layer (10% sucrose solution) was 5.4 mg / mL , the middle layer (30% sucrose solution) had a protein content of 3.9 mg / mL, and the lower layer (50% sucrose solution) had a protein content of 1.2 mg / mL. In this way, the sucrose solutions in the upper layer, middle layer and ...

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Abstract

The invention relates to a wheat gluten protein separation and purification processing technology, and discloses a method for separating wheat alcohol-soluble protein by utilizing a sucrose density gradient centrifugation method. Firstly, sucrose is dissolved in water to prepare solutions with different mass percent concentrations, then a to-be-separated alcohol-soluble protein solution soluble in70% ethanol is injected into the bottom of a centrifugal tube; the 10-70% sucrose density gradient centrifugate is prepared through the layer spreading method; centrifugation is conducted on 5000-20000 g of centrifugate for 10-60 min at the temperature below 4 DEG C, after alcohol-soluble protein settles and is balanced, a sucrose solution with the corresponding concentration is obtained, the protein content in the layer, where the protein is located, of the sucrose solution is measured under the wavelength of 280 nm by means of a spectrophotometer, and the alcohol-soluble protein can be separated into light alcohol-soluble protein, intermediate alcohol-soluble protein and heavy alcohol-soluble protein according to different sucrose densities. The method is suitable for separating and purifying the alcohol-soluble protein, results are accurate, the repeatability is good, operation is easy and feasible, and the quantity of required samples is small.

Description

technical field [0001] The invention relates to wheat gluten protein separation and purification processing technology, belongs to the field of agricultural product processing or deep processing, in particular to a method for separating or purifying wheat prolamin by means of sucrose density gradient centrifugation. Background technique [0002] Wheat prolamin has strong extensibility in the formation of gluten protein and dough, and is one of the main components that enable gluten protein to form a three-dimensional network system. However, there are currently two main classification methods for gliadins. One is to use electrophoresis to divide them into four categories according to their molecular weight: α, β, γ and ω; the other is our 2017 According to the Venn classification method proposed in 1999, it is divided into 8 kinds of proteins in 4 categories according to their solubility: single-soluble, two-soluble, three-soluble and four-soluble. At present, these classif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C07K1/36C07K1/14C07K1/34
CPCC07K14/415
Inventor 贾峰王金水陈康李桂玲王琦孙东东
Owner HENAN UNIVERSITY OF TECHNOLOGY
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