CN107652361A - Utilize the method for sucrose density gradient centrifugation separation wheat alcohol soluble protein - Google Patents
Utilize the method for sucrose density gradient centrifugation separation wheat alcohol soluble protein Download PDFInfo
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- CN107652361A CN107652361A CN201711215198.5A CN201711215198A CN107652361A CN 107652361 A CN107652361 A CN 107652361A CN 201711215198 A CN201711215198 A CN 201711215198A CN 107652361 A CN107652361 A CN 107652361A
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- soluble protein
- alcohol soluble
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
The present invention relates to wheat gluten protein separation and purifying process technology.The invention discloses a kind of method using sucrose density gradient centrifugation separation wheat alcohol soluble protein.It is first that sucrose is soluble in water, the solution of different quality percent concentration is configured to, then by the alcohol soluble protein solution for being dissolved in 70% ethanol to be separated, injection centrifuge tube bottom.10 ~ 70% sucrose density gradient centrifugation liquid is prepared using layer spread method.At 4 DEG C, with 5000 ~ 20000g, centrifuge 10 ~ 60 min, after alcohol soluble protein reaches sedimentation equilibrium, take the sucrose solution of respective concentration, using spectrophotometer under 280 nm wavelength, alcohol soluble protein, can be divided into lightweight, middle matter and heavy alcohol soluble protein by the protein content of layer sucrose solution according to different sucrose densities where measure.The present invention is applied to isolate and purify alcohol soluble protein, as a result accurately, reproducible, easy to operate, easy, needs sample size few.
Description
Technical field
The present invention relates to wheat gluten protein separation and purifying process technology, belongs to processing of farm products or intensive processing neck
Domain, particularly relate to the method using sucrose density gradient centrifugation isolated or purified wheat gliadin.
Background technology
Wheat gliadin has very strong ductility, enables mucedin shape in mucedin and dough are formed
One of main component into cubic network system.But the sorting technique of alcohol soluble protein mainly has two kinds at present, one kind is to use
The method of electrophoresis, it is divided into according to the size of molecular weight：The class of α, beta, gamma and ω etc. four；Another kind, it is that we carried in 2017
The Venn sorting techniques gone out, it is divided into according to deliquescent difference：Dan Rong, two molten, three molten and 4 class, 8 kinds of albumen such as four is molten.Mesh
Before, also it is not very perfect and careful in these sorting techniques, albumen point can not be completely covered by also having in specific production
The problem of from purifying.
The content of the invention
The present invention be exactly be directed to above-mentioned prior art in the presence of weak point and provide using sucrose density gradient from
The method of heart method separation wheat alcohol soluble protein.It is first that sucrose is soluble in water, the solution of different quality percent concentration is configured to,
Then by the alcohol soluble protein solution for being dissolved in 70% ethanol to be separated, injection centrifuge tube bottom.10 ~ 70% sugarcane is prepared using layer spread method
Sucrose density gradient centrifugate.At 4 DEG C, with 5 000 ~ 20 000 g, 10 ~ 60 min are centrifuged, it is flat to treat that alcohol soluble protein reaches sedimentation
After weighing apparatus, the sucrose solution of respective concentration is taken, using spectrophotometer under 280 nm wavelength, the egg of layer sucrose solution where measure
White matter content, alcohol soluble protein can be divided into according to different sucrose densities：Lightweight alcohol soluble protein（It is 10%-30% positioned at concentration
Sucrose solution in）, middle matter alcohol soluble protein（In the sucrose solution that concentration is 30%-50%）With heavy alcohol soluble protein（It is located at
Concentration is in 50%-70% sucrose solution）.The present invention reaches separation and purifying wheat using the method for sucrose density gradient centrifugation
Alcohol soluble protein, implementation is simple, the method for supplementing and improving wheat gliadin classification.
Embodiment
The present invention is further described below with reference to embodiment：
Embodiment 1：
The Properties of Wheat Gliadin Solution for taking the ethanol of 5 mL 70% to extract（Solution to be separated）, 50 mL centrifuge tubes bottoms are injected, respectively will
The mL of sucrose solution 10 of 20%, 40% and 60% prepared, according to layer spread method successively from centrifugation bottom of the tube injection, finally from bottom
It is followed successively by upwards：60%-40%-20%- alcohol soluble protein solution to be separated, in horizontal centrifuge, after 5000 g centrifuge 30 min,
Certain solution is drawn from 20% sucrose solution, 40% sucrose solution and 60% sucrose solution respectively, utilizes micro ultraviolet-visible
Spectrophotometer detects the content of Proteins In Aqueous Solutions at 280 nm, and it is at the middle and upper levels（20% sucrose solution）Protein content
It is middle for 6.2 mg/mL（40% sucrose solution）Protein content is 5.4 mg/mL, lower floor（60% sucrose solution）Albumen
Matter content is 2.1 mg/mL.So respectively the sucrose solution of the upper, middle and lower is drawn out, be by dialysis respectively
It can obtain lightweight, middle matter and heavy alcohol soluble protein.
Embodiment 2
The Properties of Wheat Gliadin Solution for taking the ethanol of 3 mL 70% to extract（Solution to be separated）, 50 mL centrifuge tubes bottoms are injected, respectively will
The mL of sucrose solution 10 of 10%, 30% and 50% prepared, according to layer spread method successively from centrifugation bottom of the tube injection, finally from bottom
It is followed successively by upwards：50%-30%-10%- alcohol soluble protein solution to be separated, in horizontal centrifuge, 10 000 g centrifuge 30 min
Afterwards, draw certain solution from 10% sucrose solution, 30% sucrose solution and 50% sucrose solution respectively, using it is micro it is ultraviolet-can
See that spectrophotometer detects the content of Proteins In Aqueous Solutions at 280 nm, it is at the middle and upper levels（10% sucrose solution）Protein contains
Measure as 5.4 mg/mL, centre（30% sucrose solution）Protein content is 3.9 mg/mL, lower floor（50% sucrose solution）Egg
White matter content is 1.2 mg/mL.So respectively the sucrose solution of the upper, middle and lower is drawn out, pass through dialysis respectively
It can obtain lightweight, middle matter and heavy alcohol soluble protein.
Claims (2)
1. utilize the method for sucrose density gradient centrifugation separation wheat alcohol soluble protein, it is characterised in that：The sucrose density ladder
Degree centrifugation is configured as 10%-70% using the sucrose solution of various concentrations（v/w）Concentration gradient solution, with 5 000- 20
000 g centrifugal force separate wheat gliadin component.
2. condition according to claim 1 can separate wheat gliadin, it is characterised in that：Alcohol soluble protein is in concentration
To be referred to as lightweight alcohol soluble protein in 10%-30% sucrose solution, alcohol soluble protein is in the sucrose solution that concentration is 30%-50%
Referred to as middle matter alcohol soluble protein, alcohol soluble protein are referred to as heavy alcohol soluble protein in the sucrose solution that concentration is 50%-70%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711215198.5A CN107652361A (en) | 2017-11-28 | 2017-11-28 | Utilize the method for sucrose density gradient centrifugation separation wheat alcohol soluble protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711215198.5A CN107652361A (en) | 2017-11-28 | 2017-11-28 | Utilize the method for sucrose density gradient centrifugation separation wheat alcohol soluble protein |
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| Publication Number | Publication Date |
|---|---|
| CN107652361A true CN107652361A (en) | 2018-02-02 |
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| CN201711215198.5A Pending CN107652361A (en) | 2017-11-28 | 2017-11-28 | Utilize the method for sucrose density gradient centrifugation separation wheat alcohol soluble protein |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111948589A (en) * | 2019-05-17 | 2020-11-17 | 南京林业大学 | Method for centrifugally separating ferritin nanoparticles by sucrose density gradient |
| CN115201356A (en) * | 2022-06-24 | 2022-10-18 | 华中科技大学 | High-throughput unbiased endogenous liquid-liquid phase separation protein screening method |
| CN115201356B (en) * | 2022-06-24 | 2024-06-04 | 华中科技大学 | High-flux unbiased endogenous liquid-liquid phase separation protein screening method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5580959A (en) * | 1991-12-26 | 1996-12-03 | Opta Food Ingredients, Inc. | Purification of zein from corn gluten meal |
| CN106084019A (en) * | 2016-05-01 | 2016-11-09 | 上海大学 | The separation method of corn embryosperm albuminous body |
-
2017
- 2017-11-28 CN CN201711215198.5A patent/CN107652361A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5580959A (en) * | 1991-12-26 | 1996-12-03 | Opta Food Ingredients, Inc. | Purification of zein from corn gluten meal |
| CN106084019A (en) * | 2016-05-01 | 2016-11-09 | 上海大学 | The separation method of corn embryosperm albuminous body |
Non-Patent Citations (1)
| Title |
|---|
| SVETLANA RR等: "Characterization of buckwheat seed storage protein", 《J.AGRIC.CHEM》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111948589A (en) * | 2019-05-17 | 2020-11-17 | 南京林业大学 | Method for centrifugally separating ferritin nanoparticles by sucrose density gradient |
| CN115201356A (en) * | 2022-06-24 | 2022-10-18 | 华中科技大学 | High-throughput unbiased endogenous liquid-liquid phase separation protein screening method |
| CN115201356B (en) * | 2022-06-24 | 2024-06-04 | 华中科技大学 | High-flux unbiased endogenous liquid-liquid phase separation protein screening method |
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Application publication date: 20180202 |