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Polypeptide of ascidicea and preparation thereof

A sea squirt polypeptide and sea squirt technology, applied in the field of sea squirt polypeptide and its preparation, can solve problems such as applications that have not yet been reported

Inactive Publication Date: 2006-08-16
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the patent application of sea squirts has been reported on sea squirts and fatty acids, but the extraction of sea squirt polypeptides from sea squirts and its application in the treatment of hepatitis B have not been reported yet.

Method used

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  • Polypeptide of ascidicea and preparation thereof
  • Polypeptide of ascidicea and preparation thereof
  • Polypeptide of ascidicea and preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 50Kg of fresh sea squirts, eviscerated, washed, soaked with 50kg of 75% ethanol at room temperature for 2 weeks, recovered alcohol with a rotary evaporator to obtain 1000g of extract. Dissolve 200g of the extract in 500ml of distilled water, extract with 500ml of dichloromethane, dry up the solvent to obtain 825g of the water-soluble part; then dissolve as much as possible in 500ml of distilled water, extract with the same volume of n-butanol, dry up the solvent to obtain 712g of the water-soluble part Then take 100g of the water-soluble part, put it on D-101 macroporous resin column chromatography, elute with 500ml of 50% ethanol, the flow rate is 3ml / min, and dry the solvent to obtain 14g of product; carry out silica gel column chromatography with CHCl 3 :MeOH:H 2 O ratio is 7:3:0.5 eluent gradient elution, flow rate is 3ml / min, collect thin-layer chromatography ultraviolet light to detect R f The eluent is 0.5-0.6, and the solvent is dried to obtain the crude extrac...

Embodiment 2

[0047] 50Kg of fresh sea squirts, eviscerated, washed, soaked in 50kg of 75% ethanol at room temperature for 2 weeks, recovered the alcohol with a rotary evaporator at 60°C until it was odorless, and obtained 1000g of extract. Take 200g of the extract and dissolve it with 500ml of distilled water as much as possible, extract with the same volume of chloroform, dry up the solvent to obtain 805g of the water-soluble part; then dissolve it with 500ml of distilled water as much as possible, extract with the same volume of n-butanol, and dry up the solvent to get 683g of the water-soluble part Then take 100g of the water-soluble part, put it on D-101 macroporous resin column chromatography, elute with 500ml of 50% ethanol, the flow rate is 2.5ml / min, and dry the solvent to obtain 10g of product; carry out silica gel column chromatography with CHCl 3 :MeOH:H 2 O ratio is 7:3:0.5 eluent gradient elution, flow rate is 2.5ml / min, collect thin-layer chromatography ultraviolet light dete...

Embodiment 3

[0049] 50 kg of fresh sea squirts, eviscerated, washed, soaked with 50 kg of 90% ethanol at room temperature for 2 weeks, recovered the alcohol with a rotary evaporator at 60 ° C until it was odorless, and obtained 1300 g of extract. Take 200g of the extract and dissolve it in 600ml of distilled water, extract with the same volume of chloroform, dry up the solvent to obtain 830g of water-soluble part; repeat the above extraction method 3 times, dry up the solvent to get 730g of water-soluble part; 8 Macroporous resin column chromatography, eluting with 500ml of 50% ethanol, the flow rate is 2ml / min, and the solvent is collected to obtain 15g of product; silica gel column chromatography is carried out with CHCl 3 :MeOH:H 2 O ratio is 7:3:0.5 eluent gradient elution, flow rate is 3ml / min, collect thin-layer chromatography ultraviolet light to detect R f The eluent is 0.5-0.6, and the solvent is dried to obtain the crude extract of sea squirt polypeptide, and finally, the above ...

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PUM

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Abstract

Ascidian polypeptide and its production are disclosed. The procedure is carried out by immersion depressing at normal temperature by alcohol, recovering alcohol to obtain extract, extracting by organic solvent, concentrating, separation purifying by macroporous resin chromatography, silica-gel chromatography and Sephadex LH20 chromatography, gradient eluting by eluent in proportion of CHCl3: MeOH: H2O=7:3:0.5, and collecting concentrated eluent with thin-layer chromatographic ultraviolet verification Rf=0.5~0.6. The molecular weight of ascidian polypeptide is 711.4. It is simple, has less investment, convenient control for production and various raw materials.

Description

field of invention [0001] The invention belongs to the field of biochemistry and biomedicine, and specifically relates to ascidian polypeptide and a preparation method thereof. Background technique [0002] Ascidia is a Uronordata subphylum Uronordata, and a few of the anticancer components extracted from Ascidia have entered the clinical or preclinical stage. Studies have found that sea squirts contain dozens of compounds such as alkaloids, peptides, indoles, heavy metal chelating agents, polysulfides, macrolides, terpenes, etc., which have strong antitumor, antiviral, antibacterial, Inducing sarcoplasmic reticulum to release calcium, inhibiting calmodulin activity and other physiological activities, especially anti-tumor research reports (Li Zhijun, Xue Changhu, Wang Changhai. Anti-tumor bioactive substances in sea squirts. Haiyang Bulletin, 2004, 23 (5): 82-86; Ji Yubin, Chi Wenjie. Research on anti-tumor active substances in sea squirts. Journal of Harbin University of ...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K1/14
Inventor 万新祥胡文军曾凡林王瑞
Owner SOUTHERN MEDICAL UNIVERSITY
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