Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

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
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 have carried out a large number of studies on protein extraction methods from tea tree leaves, but the results are not good. The number of extracted proteins is limited to only about 2000 protein spots, which is far from meeting the needs of proteomics research.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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
Comparison scheme
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...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com