Extraction method of fructus momordicae leaf total protein for proteomics analysis

A technology of proteomics and extraction method, which is applied in the field of extraction of total protein from Luo Han Guo leaves, can solve the problem that plant leaves cannot effectively remove polyphenols and salts, achieve low pollution, high protein purity, and improve extraction efficiency Effect

Inactive Publication Date: 2015-09-02
GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for extracting the total protein of Luo Han Guo leaves for proteomic analysis, to solve the problem that polyphenols and salts cannot be effectively removed when the traditional phenol extraction method extracts plant leaf protein, and realizes The high-efficiency extraction of the total protein from the leaves of Luo Han Guo results in high protein purity, which can be applied to proteomic analysis and provides at least the advantages described later

Method used

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  • Extraction method of fructus momordicae leaf total protein for proteomics analysis

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Embodiment 1

[0029] The invention provides a method for extracting the total protein of Luo Han Guo leaves for proteomics analysis,

[0030] Mix the pretreated Luo Han Guo leaf powder with the first protein extract, let it stand at -20°C for 10 hours, centrifuge at 4°C for 30 minutes at a speed of 12000r / min, and remove the lower layer of sediment to obtain the first precipitate, wherein the The addition amount of the first protein extract is to add 2ml per 1g of Luo Han Guo leaf powder, and utilize the first protein extract to effectively crush the cells of Luo Han Guo leaf to obtain proteins, carbohydrates, lipids, organelles, pigments, sterols, etc., and 12000r / Centrifugation at a high speed of 1 min can effectively separate proteins and lipids. Proteins are mainly concentrated in the sediment layer, while most carbohydrates, lipids, pigments, sterols and other substances are dissolved in the first protein extract, that is, Dissolved in the supernatant and taken away; if the amount of ...

Embodiment 2

[0045] The invention provides a method for extracting the total protein of Luo Han Guo leaves for proteomics analysis,

[0046] Mix the pretreated Luo Han Guo leaf powder with the first protein extract, let it stand at -20°C for 12h, centrifuge at 4°C for 30min at a speed of 12000r / min, remove the lower layer of sediment to obtain the first precipitate, wherein the The amount of the first protein extract is to add 3ml per 1g of Luo Han Guo leaf powder, and the first protein extract is used to effectively break the cells of the Luo Han Guo leaf to obtain proteins, carbohydrates, lipids, organelles, pigments, sterols, etc., and 12000r / Centrifugation at a high speed of 1 min can effectively separate proteins and lipids. Proteins are mainly concentrated in the sediment layer, while most carbohydrates, lipids, pigments, sterols and other substances are dissolved in the first protein extract, that is, Dissolved in the supernatant and taken away; if the amount of the first protein e...

Embodiment 3

[0061] The invention provides a method for extracting the total protein of Luo Han Guo leaves for proteomics analysis,

[0062] Mix the pretreated Luo Han Guo leaf powder with the first protein extract, let it stand at -20°C for 14h, centrifuge at 4°C for 30min at a speed of 12000r / min, remove the lower layer of sediment to obtain the first precipitate, wherein the The addition amount of the first protein extract is to add 4ml per 1g of Luo Han Guo leaf powder, utilize the first protein extract to effectively crush the cells of Luo Han Guo leaf, obtain proteins, carbohydrates, lipids, organelles, pigments, sterols, etc., and 12000r / Centrifugation at a high speed of 1 min can effectively separate proteins and lipids. Proteins are mainly concentrated in the sediment layer, while most carbohydrates, lipids, pigments, sterols and other substances are dissolved in the first protein extract, that is, It dissolves in the supernatant and is taken away; if the amount of the first prot...

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Abstract

The invention discloses an extraction method of fructus momordicae leaf total protein for proteomics analysis, which comprises the following steps: mixing pretreated fructus momordicae leaf powder with a first protein extracting solution, standing at -20 DEG C for 10-14 hours, centrifuging at 4 DEG C for 30 minutes at the rate of 12000 r / min, and taking the understratum precipitate to obtain a first precipitate. The preparation method of the first protein extracting solution comprises the following steps: mixing acetone and beta-mercaptoethanol in a volume ratio of 10000:(5-10), adding trichloroacetic acid, mixing and storing at -20 DEG C for later use. The method has the advantages of high total protein extraction efficiency, time saving and low protein degradability, solves the problem that the traditional phenol extraction process for extracting the plant leaf protein can not effectively remove polyphenols and salts, and implements high-efficiency extraction of the fructus momordicae leaf total protein. The obtained protein has high purity, and is suitable for proteomics analysis.

Description

technical field [0001] The invention belongs to the field of protein extraction from Luo Han Guo leaves, in particular to a method for extracting total protein from Luo Han Guo leaves for proteomics analysis. Background technique [0002] Luo Han Guo (Siraidia grosvenorii (Swingle) C. Jeffrey) is a dioecious plant of Cucurbitaceae (Cucurbitaceae) and has important medical and health value. Luo Han Guo fruit has high nutritional value and is rich in fructose, glucose, protein and multivitamins. It is cool in nature and sweet in taste. Mogroside has high sweetness, low calorie, and non-toxicity. It can be used as an ideal food additive for obese patients, diabetics, and hypertensive patients. It has been recorded in the previous "Chinese Pharmacopoeia" that it has the effects of clearing away heat and moistening the lungs, smoothing the intestines and laxatives, and is used to treat dry cough due to lung fire. According to modern pharmacological research, its pharmacological...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/36C07K1/30G01N1/28
Inventor 潘丽梅付强韦荣昌马小军白隆华黄芩芬
Owner GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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