CN1443852A - Extraction method of narcissus seed glycopeptide - Google Patents
Extraction method of narcissus seed glycopeptide Download PDFInfo
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- CN1443852A CN1443852A CN 02110169 CN02110169A CN1443852A CN 1443852 A CN1443852 A CN 1443852A CN 02110169 CN02110169 CN 02110169 CN 02110169 A CN02110169 A CN 02110169A CN 1443852 A CN1443852 A CN 1443852A
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Abstract
The method for extracting narcissus seed glycopeptide includes the following steps: coagulating protein being in narcissus seed and making it into cured narcissus seed, adding water and pulping to obtain pulp liquor, making enzymolysis to obtain liquid eymolyte, inactivating the liquid zymolyte, making centrifugal separation to obtain supernatant fluid, treating supernatant fluid, vacuum concentrating to remote water content and obtain the concrete, centrifugal removing dregs, precipitating the supernatant fluid with ethyl alcohol, separating out precipitate, dissolving the precipitate in water, centrifugal removing impurity from obtained aqueous solution, spray-drying at 120-170 deg.C so as to obtain narcissue glycopeptide.
Description
Technical field
The present invention relates to a kind of method of from Semen Narcissi Chinensis, extracting glycopeptide.
Background technology
Occurring in nature plant, microorganism, animal all contain polysaccharide, and outside existing divided by free state, the form that also can combine with protein exists.Sugar becomes glycoprotein with protein bound, is the structural material of some histoorgans.The at present common chondroitin sulfate that the achyranthes bidentata glycopeptide that extracts from the root of bidentate achyranthes, the heparin that extracts from the small intestine of pig is arranged, from the hog snout laryngeal cartilage, extract etc.Also put down in writing the Lycium chinense glycopeptide that from wolfberry fruit, extracts in the patent documentation.Because raw material sources difference, the biological activity of extract are also different.Be subjected to the different restriction of raw material sources simultaneously, none is unified the extraction process of standard.Existent method is according to Material Characteristics, adopts the mode that is soaked in water behind the raw material crushing is become water-soluble peptide and amino acid with the proteolysis in its intravital glycoprotein, reaches the purpose of extraction then with organic solvent deposit.
Semen Narcissi Chinensis claims Chrysomyia megacephala (Fabricius), engineering fly maggot again, and extracting chitosan from Semen Narcissi Chinensis has prior art.But do not find from Semen Narcissi Chinensis, to extract the record of glycopeptide at present.
Summary of the invention
Technical problem to be solved by this invention provides the extracting method of narcissus seed glycopeptide, and is simple, extraction yield is high, non-environmental-pollution.
The extracting method of narcissus seed glycopeptide of the present invention is characterized in that the protein coagulating in the Semen Narcissi Chinensis is made the boiled water fairy maiden, add water and pull an oar into slurries, carry out enzymolysis then and get enzymolysis solution, the deactivation enzymolysis solution, centrifugation obtains supernatant liquor, then supernatant liquor is handled.
The present invention makes the insoluble sugar-protein in the Semen Narcissi Chinensis be converted into solubility glycopeptide, free peptide, amino acid, efficient height by the mode of enzymolysis.
Semen Narcissi Chinensis forms earlier the boiled water fairy maiden, makes protein coagulated method, can be according to the multiple mode of prior art, and the most common is method by boiling.Concrete technology can be preferred: the water of Semen Narcissi Chinensis and 1～2 times of weight boiled under 100 ℃ 5～10 minutes.
Boiled water fairy maiden water is pulled an oar into slurries, and the water of preferred boiled water fairy maiden and 1～3 times of weight is pulled an oar.Slurries are 7～7.5 at PH, temperature is that enzymolysis obtained enzymolysis solution in 3～5 hours under 40～45 ℃, neutral protease, and the deactivation enzymolysis solution makes the enzyme termination reaction then.The deactivation enzymolysis solution carries out centrifugation, obtains supernatant liquor and slag, isolates supernatant liquor.When enzymolysis, neutral protease can adopt suitable proteolytic enzyme, preferred 1398 neutral proteinase.
The supernatant liquor that more than obtains, its treating processes comprise with organic solvent and precipitating, and throw out are dissolved purification, purification liquid drying.Provide following a kind of concrete operation steps:
Supernatant liquor removes water through vacuum concentration and becomes concentrated solution, centrifugal slagging-off, and the clear liquid ethanol sedimentation is isolated precipitation, and precipitation is used water dissolution, the centrifugal again removal of impurities of the aqueous solution that obtains, clear liquid obtains narcissus seed glycopeptide 120～170 ℃ of following spraying dryings.
The concentration of described concentrated solution is 20～25%.Used concentration of ethanol is best 〉=and 95%, sedimentation time was at least 12 hours.Also can adopt other organic solvent, as other alcohols, acetone.Consumption of ethanol can be selected according to the clear liquid before the precipitation: ethanol=1: 4～5 (volume ratio), amount of alcohol is more more not to influence whole technology.Can suitably heat for the ease of dissolving during resolution of precipitate.
The narcissus seed glycopeptide that obtains by above-mentioned technology, wherein glycoprotein content 75～97%, glycoprotein is the integrated structure (glycopeptide) of polysaccharide and peptide, molecular weight is 2600～4200, wherein polysaccharide is not less than 25.0% of glycopeptide (by dry product) total amount in dextrose anhydrous, and peptide is not less than 50% of glycopeptide (by dry product) total amount.Glycoprotein is the main effect place of pharmacodynamics.In addition, also has the not impurity of principal component of the parts such as substituting group that may exist in moisture, sodium ion and the saccharide residue in the glycopeptide.
Other quality index is as follows:
Proterties: be brown or pale brown look amorphous powder, gas is little, and it is special to distinguish the flavor of, have draw moist.This product is easily molten in water, and is insoluble in ethanol, acetone, chloroform.
Fastness to alkali: PH5.5～7.5
Weight loss on drying:＜6.0%
Ignition residue:＜13%
Heavy metal:＜20ppm
Arsenic salt:＜2ppm
Through pharmacodynamic study, narcissus seed glycopeptide has to be regulated body's immunity significantly, suppress tumor growth, share the synergy that alleviates its toxic side effect and strengthen tumor killing effect with chemotherapeutic.
Advantage of the present invention:
Extract glycopeptide from Semen Narcissi Chinensis, technology is simple, and extraction yield height, yield are up to 2.8%, and leaching process does not have the three wastes and produces, and has avoided environmental pollution.
Description of drawings
Fig. 1 is the efficient gel color atlas of narcissus seed glycopeptide.
Embodiment
The water of 10kg Semen Narcissi Chinensis and 1.5 times of weight boiled under 100 ℃ 5 minutes, obtain the boiled water fairy maiden, mix with the water of 2 times of weight and pull an oar into slurries, slurries are 7 at PH, temperature is that enzymolysis obtained enzymolysis solution in 3 hours under 45 ℃, 1398 neutral proteinase, and 90 ℃ of following deactivations obtain the deactivation enzymolysis solution.The deactivation enzymolysis solution carries out centrifugation, removes slag, obtains supernatant liquor.Supernatant liquor becomes concentrated solution at 60 ℃ of following vacuum concentration, and concentration is 20～25%, centrifugal slagging-off, and clear liquid is with 95% ethanol sedimentation 12 hours, clear liquid: ethanol=1: 5 (volume ratio).Quiescent settling is isolated precipitation, and precipitation is used water dissolution, the centrifugal again removal of impurities of the aqueous solution that obtains, and clear liquid obtains narcissus seed glycopeptide 250 grams 120 ℃ of following spraying dryings.
The above-mentioned narcissus seed glycopeptide that obtains, glycoprotein content are 90%, and wherein polysaccharide is 28%, peptide 62%.
By the color atlas of Fig. 1, go out the place, peak at sample and have only an independent peak shape and do not have other peak shape appearance, can infer that narcissus seed glycopeptide is the one-piece construction that polysaccharide and peptide link together.
Claims (9)
1, the extracting method of narcissus seed glycopeptide is characterized in that the protein coagulating in the Semen Narcissi Chinensis is made the boiled water fairy maiden, adds water and pulls an oar into slurries, and carry out enzymolysis then and get enzymolysis solution, the deactivation enzymolysis solution, centrifugation obtains supernatant liquor, then supernatant liquor is handled.
2, extracting method according to claim 1 is characterized in that wherein protein coagulating of the method for Semen Narcissi Chinensis by boiling.
3, extracting method according to claim 2 is characterized in that the water of Semen Narcissi Chinensis and 1～2 times of weight boiled 5～10 minutes under 100 ℃.
4, extracting method according to claim 1 is characterized in that slurries are 7～7.5 at PH, temperature is enzymolysis 3～5 hours under 40～45 ℃, neutral protease.
5, extracting method according to claim 4 is characterized in that described neutral protease is a 1398 neutral proteinase.
6, extracting method according to claim 1, it is characterized in that treating processes to supernatant liquor comprises with organic solvent precipitates, and throw out is dissolved purification, purification liquid drying.
7,, it is characterized in that supernatant liquor is a concentrated solution through vacuum concentration, centrifugal slagging-off according to claim 1 or 6 described extracting method, the clear liquid ethanol sedimentation is isolated precipitation, the precipitation water dissolution, the aqueous solution that obtains carries out centrifugal removal of impurities, and clear liquid is 120～170 ℃ of following spraying dryings.
8, extracting method according to claim 7, the concentration that it is characterized in that described concentrated solution is 20～25%.
9, extracting method according to claim 7 is characterized in that described concentration of ethanol 〉=95%, and sedimentation time is no less than 12 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02110169 CN1443852A (en) | 2002-03-11 | 2002-03-11 | Extraction method of narcissus seed glycopeptide |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 02110169 CN1443852A (en) | 2002-03-11 | 2002-03-11 | Extraction method of narcissus seed glycopeptide |
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| CN1443852A true CN1443852A (en) | 2003-09-24 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103397068A (en) * | 2013-08-15 | 2013-11-20 | 河北大学 | Preparation method of fly maggot peptides |
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2002
- 2002-03-11 CN CN 02110169 patent/CN1443852A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103397068A (en) * | 2013-08-15 | 2013-11-20 | 河北大学 | Preparation method of fly maggot peptides |
| CN103397068B (en) * | 2013-08-15 | 2015-03-04 | 河北大学 | Preparation method of fly maggot peptides |
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