CN1702076A - Process for extracting cyclic adenosine monophosphate (cAMP) from Chinese date - Google Patents
Process for extracting cyclic adenosine monophosphate (cAMP) from Chinese date Download PDFInfo
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- CN1702076A CN1702076A CN 200410018617 CN200410018617A CN1702076A CN 1702076 A CN1702076 A CN 1702076A CN 200410018617 CN200410018617 CN 200410018617 CN 200410018617 A CN200410018617 A CN 200410018617A CN 1702076 A CN1702076 A CN 1702076A
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Abstract
The invention provides a process for extracting adenosine 3, 5-monophosphate (cAMP). The invention comprises the following steps: cleaning, airing and stoning jujube; steeping jujube for 24-28 h in water; treating supernatant in ion exchange column for several times; treat it with separation, purification and freeze drying to generate dry powder. Compared with existing technology, the invention has the advantages of low cost and non-toxicity.
Description
Technical field
The invention provides the technology of from jujube, extracting cyclic monophosphate (cAMP)
Background technology
The cyclic monophosphate (cAMP) of manufacturer production is the synthetic product both at home and abroad at present, its raw material is Yeast Nucleic Acid (RNA), it is made up of adenine nucleotide, guanylic acid, uridylate, cytidylic acid(CMP), wherein contains VITAMIN B4, ribose and phosphoric acid in the molecule of adenine nucleotide.Also contain VITAMIN B4, ribose and phosphoric acid in the molecule of cyclic monophosphate (cAMP), constitute two ester bonds by 3 ' and 5 ' two hydroxyl of phosphate and ribose and form ring texture, thereby must be called cyclic monophosphate (cAMP).Because Yeast Nucleic Acid (RNA) contains the raw material of synthetic cyclic monophosphate (cAMP), therefore at present both at home and abroad producer all adopts Yeast Nucleic Acid (RNA) to do raw material to prepare cyclic monophosphate (cAMP) by following two key steps: 1. Yeast Nucleic Acid (RNA) hydrolysis repeatedly: with in Yeast Nucleic Acid (RNA) molecule by 3 ', four kinds of Nucleotide that 5 ' phosphodiester bond connects are hydrolyzed into four kinds of isolating Nucleotide repeatedly with catalyzer, this step catalyst system therefor must have specificity, can separate four kinds of nucleotide hydrolysis, unlikely again excessive hydrolysis produces more small-molecule substance, so difficulty is big and cost is high; 2. adopt chemical process to make 3 ' and 5 ' two hydroxyl of phosphate and ribose constitute two ester bonds and form ring texture.
The problem that existing technology exists:
1. cost height: owing to use Yeast Nucleic Acid (RNA) as the raw material of producing cyclic monophosphate (cAMP), Yeast Nucleic Acid (RNA) itself is just very expensive, per kilogram is about 25000 yuan and wherein adenine nucleotide only accounts for 1/4 of Yeast Nucleic Acid (RNA), so at present domestic only several families adopt Yeast Nucleic Acid (RNA) be cyclic monophosphate (cAMP) per kilogram of raw material production at least more than 120,000 yuan, abroad producer such as Sigma company per kilogram cyclic monophosphate (cAMP) are roughly equal to 178.5 ten thousand yuan of Renminbi up to 21.5 ten thousand dollars.
2. toxic side effect is big: owing to added some non-natural catalyzer and chemical reagent in hydrolytic process and the cyclisation step repeatedly at Yeast Nucleic Acid (RNA), make and be mixed with small amount of impurities in the final product, enter and cause certain toxic side effect in the human body, that some impurity even be detained for a long time becomes in the body is carcinogenic, teratogenesis, mutagenic matter, therefore the cyclic monophosphate of producing both at home and abroad at present (cAMP) only is used for biological chemistry, molecular biology research as using in biochemical reagents use and the experimentation on animals treatment, and Shang Weijian is used for the clinical report of patient.
Summary of the invention
The production technique that the purpose of this invention is to provide the cyclic monophosphate that a kind of production cost is low, the product pure natural is without any side effects (cAMP).
The present invention extracts cyclic monophosphate (cAMP) from jujube technology realizes according to following program:
Soak: jujube is weighed after clean, airing, stoning, adds 5 liters of ratios of distilled water in 500 grams, and adding distil water soaked 24-48 hour;
Homogenate: smash above-mentioned soak solution to pieces homogenate with tissue mashing machine or tissue refiner, each continuous 3 minutes, static 20 minutes, to carry out repeatedly 3 times, each step was all carried out under 4 ℃ ± 1 ℃ temperature condition after this step reached;
Centrifugal: is 12000g centrifugal 15 minute with high speed freezing centrifuge at rotating speed with above-mentioned homogenate; Taking-up supernatant liquor, the throw out after the taking-up supernatant liquor are pressed condition homogenate and centrifugal by condition under the centrifugal item under the homogenate item, supernatant liquor and last time merging with 2.5 liters of distilled water again;
Concentrate: the supernatant liquor after will merging evaporates with thin-film evaporator or water-bath, and making its last volume is about 1000ml;
Column chromatography for the first time: the supernatant liquor after will be above-mentioned concentrated carries out ion-exchange chromatography by anion-exchange resin column; Begin to elute after dark-brown liquid discards, collect the light yellow liquid that elutes with 0.05N formic acid with a little distilled water.
Adjust potential of hydrogen (pH): with the light yellow liquid of above-mentioned collection 1N HN
4OH transfers pH to neutral (pH=7);
Column chromatography for the second time: above-mentionedly be neutral elutriant, elutriant and collecting with column chromatography for the first time for the second time by the anion-exchange resin column chromatography;
Alumina column chromatography: for the second time the column chromatography elutriant is by alumina column chromatography, discards behind the brown liquid with the Tris buffer solution elution with a little distilled water wash-out earlier and collects elutriant;
The 4th column chromatography: above-mentioned elutriant is for the third time by the anion-exchange resin column chromatography, and elutriant and collection thereof are with the 2nd column chromatography;
Lyophilize: above-mentioned elutriant is become cyclic monophosphate (cAMP) lyophilized powder with the freeze drier lyophilize.
The advantage that the present invention compared with prior art has:
1. with low cost: because this technology is extracted cyclic monophosphate (cAMP) raw materials used is jujube, as everyone knows, jujube contains the cyclic monophosphate (cAMP) of high level, with Mare Tranquillitatis jujube is example, its content is 749nmol/g, calculate by 70% yield, cyclic monophosphate (cAMP) cost of per kilogram 95% purity is about 50,000 yuans, and its cost is starkly lower than the cyclic monophosphate (cAMP) of synthetic method preparation;
2. have no side effect: the zizyphus integration of drinking and medicinal herbs, not only can be as fresh and dried fruit edible but also can be used as Chinese medicinal materials and be used as medicine, therefore the cyclic monophosphate (cAMP) that extracts from jujube belongs to all-natural product, has no side effect.
Description of drawings
The present invention does not have accompanying drawing
Embodiment
Embodiment 1, and the technology that is raw material production cyclic monophosphate (cAMP) with the regional Golden jujube in Mare Tranquillitatis realizes according to following program:
Soak: Golden jujube is weighed after clean, airing, stoning, adds 5 liters of ratios of distilled water in 500 grams, and adding distil water soaked 24-48 hour;
Homogenate: smash above-mentioned soak solution to pieces homogenate with tissue mashing machine or tissue refiner, each continuous 3 minutes, static 20 minutes, to carry out repeatedly 3 times, each step was all carried out under 4 ℃ ± 1 ℃ temperature condition after this step reached;
Centrifugal: is 12000g centrifugal 15 minute with high speed freezing centrifuge at rotating speed with above-mentioned homogenate; Taking-up supernatant liquor, the throw out after the taking-up supernatant liquor are pressed condition homogenate and centrifugal by condition under the centrifugal item under the homogenate item, supernatant liquor and last time merging with 2.5 liters of distilled water again;
Concentrate: the supernatant liquor after will merging evaporates with thin-film evaporator or water-bath, and making its last volume is about 1000ml;
Column chromatography for the first time: the supernatant liquor after will be above-mentioned concentrated carries out ion-exchange chromatography by anion-exchange resin column; Begin to elute after dark-brown liquid discards, collect the light yellow liquid that elutes with 0.05N formic acid with a little distilled water.
Adjust potential of hydrogen (pH): with the light yellow liquid of above-mentioned collection 1N HN
4OH transfers pH to neutral (pH=7);
Column chromatography for the second time: above-mentionedly be neutral elutriant, elutriant and collecting with column chromatography for the first time for the second time by the anion-exchange resin column chromatography;
Alumina column chromatography: with above-mentioned elutriant by alumina column chromatography, discard brown liquid with a little distilled water wash-out earlier after, with the Tris buffer solution elution and collect elutriant;
The 4th column chromatography: above-mentioned elutriant is for the third time by the anion-exchange resin column chromatography, and elutriant and collection thereof are with the 2nd column chromatography;
Lyophilize: above-mentioned elutriant is become cyclic monophosphate (cAMP) lyophilized powder with the freeze drier lyophilize.
Claims (1)
1, extract cyclic monophosphate (cAMP) technology from jujube, it is characterized in that: the present invention extracts cyclic monophosphate (cAMP) from jujube technology realizes according to following program:
Soak: jujube is weighed after clean, airing, stoning, adds 5 liters of ratios of distilled water in 500 grams, and adding distil water soaked 24-48 hour;
Homogenate: smash above-mentioned soak solution to pieces homogenate with tissue mashing machine or tissue refiner, each continuous 3 minutes, static 20 minutes, to carry out repeatedly 3 times, each step was all carried out under 4 ℃ ± 1 ℃ temperature condition after this step reached;
Centrifugal: is 12000g centrifugal 15 minute with high speed freezing centrifuge at rotating speed with above-mentioned homogenate; Taking-up supernatant liquor, the throw out after the taking-up supernatant liquor are pressed condition homogenate and centrifugal by condition under the centrifugal item under the homogenate item, supernatant liquor and last time merging with 2.5 liters of distilled water again;
Concentrate: the supernatant liquor after will merging evaporates with thin-film evaporator or water-bath, and making its last volume is about 1000ml;
Column chromatography for the first time: the supernatant liquor after will be above-mentioned concentrated carries out ion-exchange chromatography by anion-exchange resin column; Begin to elute after dark-brown liquid discards, collect the light yellow liquid that elutes with 0.05N formic acid with a little distilled water.
Adjust potential of hydrogen (pH): with the light yellow liquid of above-mentioned collection 1N HN
4OH transfers pH to neutral (pH=7);
Column chromatography for the second time: above-mentionedly be neutral elutriant, elutriant and collecting with column chromatography for the first time for the second time by the anion-exchange resin column chromatography;
Alumina column chromatography: for the second time the column chromatography elutriant is by alumina column chromatography, discards behind the brown liquid with the Tris buffer solution elution with a little distilled water wash-out earlier and collects elutriant;
The 4th column chromatography: above-mentioned elutriant is for the third time by the anion-exchange resin column chromatography, and elutriant and collection thereof are with the 2nd column chromatography;
Lyophilize: above-mentioned elutriant is become cyclic monophosphate (cAMP) lyophilized powder with the freeze drier lyophilize.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100186172A CN100497366C (en) | 2004-01-15 | 2004-01-15 | Process for extracting cyclic adenosine monophosphate (cAMP) from Chinese date |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100186172A CN100497366C (en) | 2004-01-15 | 2004-01-15 | Process for extracting cyclic adenosine monophosphate (cAMP) from Chinese date |
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| Publication Number | Publication Date |
|---|---|
| CN1702076A true CN1702076A (en) | 2005-11-30 |
| CN100497366C CN100497366C (en) | 2009-06-10 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB2004100186172A Expired - Fee Related CN100497366C (en) | 2004-01-15 | 2004-01-15 | Process for extracting cyclic adenosine monophosphate (cAMP) from Chinese date |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100425620C (en) * | 2006-11-09 | 2008-10-15 | 常月梅 | Technology of extracting adenosin phosphoric acid from date |
| CN102268055A (en) * | 2010-06-04 | 2011-12-07 | 南京工业大学 | Method for separating adenosine cyclophosphate |
| WO2011153872A1 (en) * | 2010-06-04 | 2011-12-15 | 南京工业大学 | Arthrobacter strain used for producing cyclic adenosine monophosphate by fermentation and use thereof |
| WO2011153874A1 (en) * | 2010-06-04 | 2011-12-15 | 南京工业大学 | Crystallization process of cyclic adenosine 3',5'-monophosphate |
| CN102875625A (en) * | 2012-10-29 | 2013-01-16 | 北京林业大学 | Extraction method of cyclic adenosine monophosphate (cAMP) with antiallergic activity from Chinese date |
| CN104761606A (en) * | 2014-01-07 | 2015-07-08 | 中国科学院兰州化学物理研究所 | Method for extracting cyclic adenosine monophosphate from jujube |
| CN105249086A (en) * | 2015-11-13 | 2016-01-20 | 侯彦国 | Preparation process of red date condensed juice high in cAMP content |
| CN105647995A (en) * | 2016-02-29 | 2016-06-08 | 山东大学 | Method for extracting 2',3'-cNMPs |
-
2004
- 2004-01-15 CN CNB2004100186172A patent/CN100497366C/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100425620C (en) * | 2006-11-09 | 2008-10-15 | 常月梅 | Technology of extracting adenosin phosphoric acid from date |
| CN102268055A (en) * | 2010-06-04 | 2011-12-07 | 南京工业大学 | Method for separating adenosine cyclophosphate |
| WO2011153872A1 (en) * | 2010-06-04 | 2011-12-15 | 南京工业大学 | Arthrobacter strain used for producing cyclic adenosine monophosphate by fermentation and use thereof |
| WO2011153874A1 (en) * | 2010-06-04 | 2011-12-15 | 南京工业大学 | Crystallization process of cyclic adenosine 3',5'-monophosphate |
| CN102268055B (en) * | 2010-06-04 | 2014-09-24 | 南京工业大学 | Method for separating adenosine cyclophosphate |
| CN102875625A (en) * | 2012-10-29 | 2013-01-16 | 北京林业大学 | Extraction method of cyclic adenosine monophosphate (cAMP) with antiallergic activity from Chinese date |
| CN102875625B (en) * | 2012-10-29 | 2015-07-08 | 北京林业大学 | Extraction method of cyclic adenosine monophosphate (cAMP) with antiallergic activity from Chinese date |
| CN104761606A (en) * | 2014-01-07 | 2015-07-08 | 中国科学院兰州化学物理研究所 | Method for extracting cyclic adenosine monophosphate from jujube |
| CN105249086A (en) * | 2015-11-13 | 2016-01-20 | 侯彦国 | Preparation process of red date condensed juice high in cAMP content |
| CN105647995A (en) * | 2016-02-29 | 2016-06-08 | 山东大学 | Method for extracting 2',3'-cNMPs |
| CN105647995B (en) * | 2016-02-29 | 2019-03-08 | 山东大学 | A kind of method for extracting 2 ', 3 '-cyclic nucleoside monophosphates |
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| Publication number | Publication date |
|---|---|
| CN100497366C (en) | 2009-06-10 |
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