Method for extracting total protein of momordica grosvenori leaf for proteomic 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 of inability to effectively remove polyphenols and salts, and achieve the effects of less pollution, improved extraction efficiency, and high-efficiency extraction

Inactive Publication Date: 2019-05-10
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the above-mentioned technical problems, the present invention provides a method for extracting the total protein of Luo Han Guo leaves for proteomic analysis. The method of the present invention can solve the problem that the traditional phenol ext

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0028] Example 1

[0029] S1: Cut the Luo Han Guo leaves into pieces, add liquid nitrogen and polyvinylpyrrolidone, and grind; After grinding the Luo Han Guo leaves, mix them with supercritical carbon dioxide at a mass ratio of 1:16-20, and put them into the supercritical extraction tank. The extraction pressure is 18-22MPa, the extraction temperature is 30-33℃, the powder after extracting the lipids is mixed with water at a mass ratio of 1:80-120, stirring for 5-15min, adding sodium bicarbonate with a molar concentration of 0.4-0.6mol / L The solution is allowed to stand for 2-4 hours at 20-25°C until the pH value is 9.2-9.6, washed with water to a pH value of 6.8-7.1, filtered and freeze-dried to obtain Luo Han Guo leaf powder.

[0030] The pretreated Luo Han Guo leaf powder was mixed with the protein extract, left standing at -20°C for 10 hours, centrifuged at 4°C for 30 minutes, at a speed of 12000 r / min, and the lower layer of precipitation was removed. The protein extract was a...

Example Embodiment

[0036] Example 2

[0037] S1: Cut the leaves of Luo Han Guo into pieces, add liquid nitrogen and polyvinylpyrrolidone, and grind; After grinding the leaves of Luo Han Guo, mix them with supercritical carbon dioxide at a mass ratio of 1:16 and put them into a supercritical extraction tank with an extraction pressure of 18MPa. The extraction temperature is 30°C, the powder after extracting the lipids is mixed with water at a mass ratio of 1:80, stirring for 15 minutes, adding a sodium bicarbonate solution with a molar concentration of 0.4mol / L until the pH is 9.2, and standing at 20°C 4h, wash with clean water to pH 6.8, filter and freeze-dry to obtain Luo Han Guo leaf powder.

[0038] The pretreated Luo Han Guo leaf powder was mixed with the protein extract, left standing at -20°C for 10 hours, centrifuged at 4°C for 30 minutes, at a speed of 12000 r / min, and the lower layer of precipitation was removed. The protein extract was added in an amount of Add 3ml per 1g of Luo Han Guo le...

Example Embodiment

[0044] Example 3

[0045] S1: Cut the Luo Han Guo leaves into pieces, add liquid nitrogen and polyvinylpyrrolidone, and grind; After grinding the Luo Han Guo leaves, mix them with supercritical carbon dioxide at a mass ratio of 1:20, and put them into a supercritical extraction tank with an extraction pressure of 22 MPa. The extraction temperature is 33°C, the powder after extracting the lipids is mixed with water in a mass ratio of 1:120, stirring for 5 minutes, adding a sodium bicarbonate solution with a molar concentration of 0.6mol / L until the pH is 9.6, and leaving it at 25°C After 2h, wash with clean water to pH 7.1, filter, freeze-dry, and obtain Luo Han Guo leaf powder.

[0046] The pretreated Luo Han Guo leaf powder was mixed with the protein extract, left standing at -20°C for 10 hours, centrifuged at 4°C for 30 minutes, at a speed of 12000 r / min, and the lower layer of precipitation was removed. The protein extract was added in an amount of Add 4ml per 1g of Luo Han Guo...

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Abstract

The invention relates to the field of biology, and discloses a method for extracting a total protein of a momordica grosvenori leaf for proteomic analysis. The method comprises steps of S1 mixing pretreated momordica grosvenori leaf powder with a protein extract, standing, centrifuging, and taking a lower layer precipitate; S2 freezing and drying the precipitate, pulverizing to obtain precipitatepowder, and storing for prepare use; S3 mixing the precipitate powder with a protein lysis solution, and conducting pyrolysis to obtain the momordica grosvenori leaf lysis solution; S4 centrifuging the momordica grosvenori leaf lysis solution, taking a supernatant, and drying to obtain the total protein of the momordica grosvenori leaf. The method can solve the problem that a traditional phenol extraction method cannot effectively remove polyphenols and salt substances when a plant leaf protein is produced, the high-efficiency extraction of the total protein of the momordica grosvenori leaf isachieved, the protein purity is high, and the method can be applied to the proteomics analysis.

Description

technical field [0001] The present invention relates to the field of biology, in particular to a method for extracting total protein from Luo Han Guo leaves for proteomics analysis Background technique [0002] Luo Han Guo belongs to Cucurbitaceae and is a dioecious plant of the genus Luo Han Guo, which 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. Luo Han Guo is high in sweetness, low in calories, and non-toxic. 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 active ingredient is mainly Monk Fruit Sap...

Claims

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Application Information

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IPC IPC(8): C07K1/14
CPCY02P20/54
Inventor 朱诚胡铭周杨慧周莲王秉
Owner ZHEJIANG SCI-TECH UNIV
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