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Peach bud protein extracting method

An extraction method and protein technology, applied in the field of plant protein extraction, can solve the problems of poor quality of peach blossom bud protein, inability to effectively remove polyphenols and salts, inability to separate by proteomics, and the like, and the method and steps are simple. , Easy to remove impurities, high protein purity effect

Inactive Publication Date: 2019-03-08
CHANGLI INST OF POMOLOGY HEBEI ACADEMY OF AGRI & FORESTRY SCI
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  • Abstract
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Problems solved by technology

[0003] The traditional phenol method is used to extract protein from plant leaves. Taking advantage of the fact that polysaccharides are insoluble in the phenol phase, saturated phenol can be used to extract proteins, thereby removing polysaccharides and other phenol-insoluble substances in the sample. However, the phenol method Cannot effectively remove polyphenols and salts
Therefore, the quality of the peach bud protein extracted by the phenol method is not good, and it cannot be used for the next step of proteomic separation, and thus cannot be used for proteomic analysis

Method used

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Embodiment Construction

[0016] The invention provides an extraction method suitable for peach flower bud protein. The method includes the following steps:

[0017] Step 1. Picking peach blossom buds in winter, peeling off the dry scales of the peach blossom buds, freezing the peach blossom buds with liquid nitrogen and storing them in a refrigerator at -70°C;

[0018] Step 2. Weigh 1g of peach blossom buds stored in the refrigerator, grind the weighed peach blossom buds into powder in liquid nitrogen with a mortar, add 0.1g of PPVP during the grinding process, and weigh 0.5g of peach blossom bud powder as a sample. ;

[0019] Step 3. Take a 10ml centrifuge tube and add 4.5ml of -20°C pre-chilled vegetable protein extract, 10ul PMSF and 10ulDTT, transfer the sample to a 10ml centrifuge tube containing the extract, vortex for 30s, and hold at 4°C. Lower extraction for 1h;

[0020] Step 4. Shake the sample well and place it in a high-speed refrigerated centrifuge, centrifuge for 15 minutes at 4°C and 8000r / mi...

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Abstract

The invention belongs to the technical field of plant protein extraction and particularly relates to a peach bud protein extracting method. The peach bud protein extracting method achieves separationand purification of proteins in peach buds through dissolution, precipitation, purification and drying. The peach bud protein extracting method is simple in process; peach bud proteins extracted through the peach bud protein extracting method are high in purity and low in content of impurities and facilitate research related to peach bud proteomics.

Description

Technical field [0001] The invention belongs to the technical field of plant protein extraction, and specifically relates to an extraction method suitable for peach flower bud protein. Background technique [0002] The difficulty of plant protein extraction is that the protein content in plant cells is low, and it also contains many organic substances that interfere with protein extraction, such as phenols, pigments, lipids and nucleic acids. The peach blossom buds are even more suitable for extracting protein from the inside due to the outer scales. The buds are very small, the nucleic acid content is high, and the phenolic substances are high. It is difficult to extract high-quality proteins for a series of experiments. However, the conventional TCA-acetone extraction method cannot obtain an ideal electrophoretic pattern, which severely restricts its proteomics. Research. Proteomics technology is a powerful tool to study the synthesis of secondary metabolites in peach blossom ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K1/34C07K1/36
CPCC07K1/145C07K1/34C07K1/36
Inventor 李永红王召元常瑞丰刘国俭陈湖徐金涛韩继成
Owner CHANGLI INST OF POMOLOGY HEBEI ACADEMY OF AGRI & FORESTRY SCI
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