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A kind of extraction method of proinflammatory protein component of litchi

An extraction method and litchi 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, high product purity, and increased mRNA expression level

Active Publication Date: 2022-02-11
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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  • A kind of extraction method of proinflammatory protein component of litchi
  • A kind of extraction method of proinflammatory protein component of litchi
  • A kind of extraction method of proinflammatory protein component of litchi

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 and crush it, add pre-cooled acetone and mix thoroughly in a homogenizer at -20°C at 20,000r / min for 10 min, filter with filter paper, and use acetone-ether (1:1v / v ) 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 through a 0.45μm filter memb...

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, crush it, add pre-cooled acetone, mix thoroughly in a homogenizer at -30°C at 15000r / min for 15 min, filter with filter paper, and use acetone-ether (2:1v / v ) 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 it through a 0.45μm filter membra...

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 10 min, filter with filter paper, and use acetone-ether (3:1v / v ) 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, pass through a 0.45 μm filter m...

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Abstract

The invention discloses a method for extracting proinflammatory protein components of litchi, which comprises the following steps: pulverizing litchi pulp, mixing with acetone; separating solid and liquid, washing the precipitate with acetone-ether mixed solution, and grinding under liquid nitrogen; The precipitate is mixed with the extract, extracted, solid-liquid separated, the supernatant is taken, and dried to obtain litchi crude protein; the sample is loaded onto a chromatographic column containing a weak anion exchange resin, and gradient eluted; the elution obtained in the previous step is obtained by using a size-exclusion chromatographic column The solution was separated, eluted with ultrapure water, and the eluate was collected and dried. The invention uses liquid nitrogen to grind and break to accelerate the dissolution of the pro-inflammatory active ingredients in the litchi, and this method extracts in a low-temperature environment and has little influence on protein activity. The experimental results showed that the litchi pro-inflammatory protein promoted a significant increase in the mRNA expression levels of inflammation-related mediators COX‑2, iNOS, and TNF‑α. The extraction of litchi proinflammatory protein components by the method of the invention has high product purity and good proinflammatory activity.

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 Patents(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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