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Method for extracting vegetable butter raft

A lipid raft and plant technology, applied in the field of plant lipid raft extraction, can solve the problems that the method of extracting plant lipid raft has not been reported yet, and achieve the effect of preventing degradation, high quality and good repeatability

Inactive Publication Date: 2011-04-13
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are only methods for the extraction and purification of plant plasma membranes, but methods for further extraction of plant lipid rafts have not been reported

Method used

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  • Method for extracting vegetable butter raft
  • Method for extracting vegetable butter raft
  • Method for extracting vegetable butter raft

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1, the extraction of lipid rafts in Arabidopsis leaves

[0042] 1) Take the leaves of Arabidopsis thaliana, and add 3ml of 50mM MOPS buffer solution with pH 7.5 per 1g of leaves;

[0043] The composition of MOPS buffer is: 50mM 3-(N-morpholino)propanesulfonic acid (MOPS), 0.33M sucrose, 10% mass percent glycerol, 5mM ethylenediaminetetraacetic acid (EDTA), 0.2% Mass percent casein, 0.6% mass percent polyvinylpyrrolidone (PVPP), 5mM dithiothreitol (DTT), 5mM ascorbic acid, the concentration of each substance is their final concentration in MOPS buffer;

[0044] 2) Polish the leaves of the above step 1) in a high-speed tissue homogenizer for 4-5 times, each time for 6-8s;

[0045] 3) Filter the crushed plant leaves in the above step 2) with four layers of gauze to obtain a supernatant, and add PMSF to the supernatant to make the final concentration 0.5mM;

[0046] 4) centrifuge the reaction liquid in the above step 3) at 10,000 g for 15 min, and collect the supe...

Embodiment 2

[0062] Example 2, the extraction of lipid rafts in spinach leaves

[0063] 1) Take spinach leaves, add 3ml of 50mM MOPS buffer solution with pH 7.5 per 1g of leaves;

[0064] The composition of MOPS buffer is: 50mM 3-(N-morpholino) propanesulfonic acid (MOPS), 0.33M sucrose, 10% by mass content glycerol, 5mM ethylenediaminetetraacetic acid EDTA, 0.2% by mass Content casein, 0.6% mass percent polyvinylpyrrolidone (PVPP), 5mM dithiothreitol (DTT), 5mM ascorbic acid, the concentration of each substance is their final concentration in MOPS buffer;

[0065] 2) Polish the leaves of the above step 1) in a high-speed tissue homogenizer for 4-5 times, each time for 6-8s;

[0066] 3) Filter the crushed plant leaves in the above step 2) with four layers of gauze to obtain a supernatant, and add PMSF to the supernatant to make the final concentration 0.5mM;

[0067] 4) centrifuge the reaction liquid in the above step 3) at 10,000 g for 15 min, and collect the supernatant;

[0068] 5) C...

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PUM

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Abstract

The invention discloses a method for extracting vegetable tallow raft. The method comprises the following steps: (1) the plasma membrane is split to obtain lysis solution; (2) the density gradient centrifugation is carried out on the lysis solution in the step (1) to obtain the plant raft. The method uses dithiothreitol (DTT) to reduce the disulfide linkage, thus causing that the protein is fullydissolved. But phenylmethylsulfonyl fluoride (PMS) can effectively prevent the degradation of the protein in the extraction process. The method for extracting the plant raft has the advantages of being fast, simple and practicable and obtaining the tallow raft with higher quality and better repeatability. The method can play important role in the research of the vegetable tallow raft and has veryhigh practical using value.

Description

technical field [0001] The invention relates to a method for extracting plant lipid rafts. Background technique [0002] Simons et al. found in 1997 that most mammalian cell plasma membranes have microdomains rich in cholesterol and sphingolipids (sphingomyelin and glycosphingolipids), called "lipid rafts". In recent years, similar membrane microdomains have been found in plants such as Arabidopsis, tobacco and alfalfa. Lipid rafts are about 70nm in size and are dynamic structures located on the outer leaflet of the plasma membrane. Since sphingomyelin has longer saturated fatty acid chains and stronger intermolecular forces, these regions have dense structures, which are between disordered liquids and liquid crystals, and are called ordered liquids (Liquid-ordered). These regions can resist the extraction of non-ionic detergents at low temperatures, so they are also called detergent-resistant membranes (DRMs). [0003] The composition and structural characteristics of li...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C11B1/00
Inventor 林金星李瑞丽陈彤
Owner INST OF BOTANY CHINESE ACAD OF SCI
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