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Litchi proinflammatory protein component extraction method

An extraction method and lychee technology, applied in the biological field, can solve the problems that restrict the research and utilization of such proteins, and achieve the effects of good pro-inflammatory activity and high product purity.

Active Publication Date: 2019-07-05
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, there is no extraction method for this type of protein, which seriously restricts the research and utilization of this type of protein.

Method used

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  • Litchi proinflammatory protein component extraction method
  • Litchi proinflammatory protein component extraction method
  • Litchi proinflammatory protein component extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1 A kind of extraction method of litchi proinflammatory protein component

[0069] 1. Experimental method

[0070] 1. Peel and remove the core of the lychee, crush it, add pre-cooled acetone, mix thoroughly with a homogenizer at -20°C at 20,000r / min for 10min, filter with filter paper, and use acetone-ether (1:1v / v) to obtain the precipitate The mixed solution was washed 3 times to obtain the precipitate;

[0071] 2. Grind the precipitate with liquid nitrogen, add 0.01mol / L potassium phosphate buffer extract (containing 0.15mol / L NaCl) at 10 times pH 7.4, shake and extract at 4°C for 3h, and centrifuge at 5000r / min for 10min to obtain the above The clear liquid was freeze-dried in vacuum into powder to obtain litchi crude protein powder.

[0072] 3. Equilibrate the ion-exchange resin DEAE Sepharose FF to room temperature, degas it, add it to the chromatographic column at one time, prepare the lychee crude protein powder into a 40mg / mL solution, pass it thro...

Embodiment 2

[0077] Embodiment 2 A kind of extraction method of litchi proinflammatory protein component

[0078] 1. Experimental method

[0079] 1. Peel and remove the core of the lychee and crush it, add pre-cooled acetone and mix thoroughly in a homogenizer at -30°C at 15000r / min for 15min, filter with filter paper, and use acetone-ether (2:1v / v) to obtain the precipitate The mixed solution was washed 2 times to obtain the precipitate;

[0080] 2. Grind the precipitate with liquid nitrogen, add 0.01mol / L potassium phosphate buffer extract (containing 0.2mol / L NaCl) at 20 times pH 7.4, shake and extract at 2°C for 2 hours, and centrifuge at 5000r / min for 15min to obtain the above The clear liquid was freeze-dried in vacuum into powder to obtain litchi crude protein powder.

[0081] 3. Equilibrate the ion exchange resin DEAE Sephadex A-25 to room temperature, degas it, add it to the chromatographic column at one time, prepare the lychee crude protein powder into a 60mg / mL solution, pass...

Embodiment 3

[0086] Embodiment 3 A kind of extraction method of litchi proinflammatory protein component

[0087] 1. Experimental method

[0088] 1. Peel and remove the core of the lychees and crush them, add pre-cooled acetone and mix thoroughly in a homogenizer at -40°C at 20,000r / min for 10min, filter with filter paper, and use acetone-ether (3:1v / v) to obtain the precipitate The mixed solution was washed 2 times to obtain the precipitate;

[0089] 2. Grind the precipitate with liquid nitrogen, add 0.01mol / L potassium phosphate buffer extract (containing 0.3mol / L NaCl) at 30 times pH 7.4, shake and extract at 6°C for 1 hour, and centrifuge at 5000r / min for 15 minutes to obtain the above The clear liquid was freeze-dried in vacuum into powder to obtain litchi crude protein powder.

[0090] 3. Equilibrate the ion exchange resin DEAE Sephadex A-50 to room temperature, degas it, add it to the chromatographic column at one time, prepare 80 mg / mL solution of litchi crude protein powder, pas...

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Abstract

The invention discloses a litchi proinflammatory protein component extraction method which includes the steps: crushing litchi pulp and blending the litchi pulp and acetone; performing solid-liquid separation, washing precipitates by acetone-ether mixed liquid and grinding the precipitates under liquid nitrogen; mixing the grinded precipitates with extraction liquid, performing extraction and solid-liquid separation and drying supernate to obtain litchi crude proteins; loading samples to a chromatographic column containing weak anion exchange resin, and performing gradient elution; separatingeluent obtained in the previous step by an exclusion chromatographic column, performing elution by ultra-pure water and collecting and drying the eluent. The precipitates are grinded and crushed by the aid of the liquid nitrogen, dissolution of proinflammatory effective components in litchis is accelerated, the components are extracted in a low-temperature environment according to the method, andthe activity of the proteins is less affected. Experimental results display that the litchi proinflammatory proteins promote remarkable increase of the mRNA expression level of inflammation-associatedmedia COX-2, iNOS and TNF-alpha. The litchi proinflammatory protein components are extracted by the method, product purity is high, and proinflammatory activity is good.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, to a method for extracting proinflammatory protein components of litchi. Background technique [0002] Litchi (Litchi chinensis SONN) belongs to the fruit of Sapinaceae Litchi, native to China. At present, my country's litchi cultivation is mainly in Guangdong, Hainan, Guangxi, Fujian, Yunnan, Taiwan and other provinces. Abroad, it is also planted in subtropical regions such as Vietnam, Laos, Sri Lanka, Indonesia, Puerto Rico, the Philippines, Mauritius, and Malagasy. Litchi has high nutritional value and various nourishing and beautifying effects, and enjoys the reputation of "the best fruit in Lingnan" and "treasure among fruits". In the "Compendium of Materia Medica" written by Li Shizhen, a great medical scientist in ancient my country, it is recorded: "Always eat lychees, which can nourish the brain and keep fit, treat miasma and swell, appetize and benefit the spleen, and d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C07K1/36C07K1/30C07K1/18
CPCC07K14/415
Inventor 赵雷陈慧芳胡卓炎王凯
Owner SOUTH CHINA AGRI UNIV
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