Efficient extraction method for proteins in tea leaves

An extraction method and protein technology, applied in the field of protein extraction, can solve the problems of poor effect, limited tea proteomics research, and influence on protein extraction and separation of tea leaves, achieving low extraction efficiency, good separation effect, and clear protein spots. Effect

Pending Publication Date: 2019-07-09
LONGYAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because tea tree leaves contain a large amount of crude fiber and secondary metabolites, the extraction and separation of tea tree leaf proteins are seriously affected, which limits further research on tea proteomics
In recent years, many scholars hav

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  • Efficient extraction method for proteins in tea leaves
  • Efficient extraction method for proteins in tea leaves
  • Efficient extraction method for proteins in tea leaves

Examples

Experimental program
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Effect test

Embodiment 1

[0029] A kind of efficient tea tree leaf protein extraction method

[0030] Preparation of TCA / acetone mixed solution A: Acetone solution is used for preparation, wherein the content of TCA is 15%, the content of mercaptoethanol is 0.1%, and the content of acetone is 84.9%.

[0031] Preparation of TCA / acetone mixed solution B: Acetone solution is used for preparation, wherein the content of TCA is 12%, the content of mercaptoethanol is 0.08%, and the content of acetone is 87.2%.

[0032] Preparation of protein sample lysate: use sterile water to prepare, including 9.5 mol / L urea, 0.8% dithiothreitol content, 3.8% CHAPS content, and 1.8% Ampholine content with a pH value of 5-8.

[0033] 1) Preliminary grinding, extraction and standing of leaves

[0034] Take tea tree leaves (two leaves and one core), cut them into 1cm flakes with scissors and mix well, weigh 4g of tea tree leaves and put them in a mortar, add about 0.08g of PVP, add liquid nitrogen, quickly grind for 8s, add ...

Embodiment 2

[0043] A kind of efficient tea tree leaf protein extraction method

[0044] Preparation of TCA / acetone mixed solution A: Acetone solution is used for preparation, wherein the content of TCA is 15%, the content of mercaptoethanol is 0.1%, and the content of acetone is 84.9%.

[0045] Preparation of TCA / acetone mixed solution B: Acetone solution is used for preparation, wherein the content of TCA is 12%, the content of mercaptoethanol is 0.08%, and the content of acetone is 87.2%.

[0046] Preparation of protein sample lysate: use sterile water to prepare, including 9.5 mol / L urea, 0.8% dithiothreitol content, 3.8% CHAPS content, and 1.8% Ampholine content with a pH value of 5-8.

[0047] 1) Preliminary grinding, extraction and standing of leaves

[0048] Take tea tree leaves (two leaves and one core), cut them into 1cm flakes with scissors and mix well, weigh 4g of tea tree leaves and place them in a mortar, add about 0.12g of PVP, add liquid nitrogen, quickly grind for 10s, add...

Embodiment 3

[0057] A kind of efficient tea tree leaf protein extraction method

[0058] Preparation of TCA / acetone mixed solution A: Acetone solution is used for preparation, wherein the content of TCA is 15%, the content of mercaptoethanol is 0.1%, and the content of acetone is 84.9%.

[0059] Preparation of TCA / acetone mixed solution B: Acetone solution is used for preparation, wherein the content of TCA is 12%, the content of mercaptoethanol is 0.08%, and the content of acetone is 87.2%.

[0060] Preparation of protein sample lysate: use sterile water to prepare, including 9.5 mol / L urea, 0.8% dithiothreitol content, 3.8% CHAPS content, and 1.8% Ampholine content with a pH value of 5-8.

[0061] 1) Preliminary grinding, extraction and standing of leaves

[0062] Take tea tree leaves (two leaves and one core), cut them into 1.5cm flakes with scissors and mix well, weigh 5g of tea tree leaves and put them in a mortar, add about 0.12g of PVP, add liquid nitrogen, quickly grind for 10s, a...

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Abstract

The invention discloses an efficient extraction method for proteins in tea leaves. Fresh tea leaves are picked, a mixed TCA/acetone solution is adopted for extraction, separation and standing are conducted, sediments after standing are further washed and purified, low-temperature vacuum freezing drying is conducted, crude proteins are obtained, then lysate is adopted for splitting decomposition ofthe crude proteins, supernatant is obtained through centrifugation, and a pure protein solution is obtained. According to the method, the quantity of the proteins obtained through extraction is effectively increased, after 2D-PAGE is adopted for separation, the separation effect is good, the proteins can be effectively separated, and the protein spots are clear; LC/MS/MS is adopted for identifying the proteins, and the result shows that there are 4,117-4,218 proteins extracted through the method. The method lays an important foundation for the research of tea leaf proteomics.

Description

technical field [0001] The invention belongs to the technical field of protein extraction, and in particular relates to an efficient method for extracting protein from tea tree leaves. Background technique [0002] Tea is one of the important economic crops in our country, and aroma is an important indicator of tea evaluation grade. Therefore, in recent years, many scholars have studied the reasons for the formation of tea aroma from different angles, hoping to increase the content of tea aroma substances and improve the quality of tea through artificial adjustment. Among them, proteomics is an important method for studying the metabolism of tea tree leaves. Using proteomics methods, it can effectively analyze the changes in the metabolic process of tea tree leaves, find key enzymes in the aroma synthesis pathway of tea tree leaves, and then regulate them. However, because tea tree leaves contain a large amount of crude fiber and secondary metabolites, the extraction and se...

Claims

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Application Information

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IPC IPC(8): C07K1/14
CPCC07K1/145
Inventor 王海斌陈晓婷王裕华张华彬丁力孔祥海
Owner LONGYAN UNIV
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