Method for extracting cyclic adenosine monophosphate and application of cyclic adenosine monophosphate

A cyclic adenosine monophosphate and the obtained technology are applied in chemical instruments and methods, preparation of sugar derivatives, sugar derivatives, etc., and can solve the problems of large waste water volume and rising waste water treatment costs, and achieve shortened production cycle and stable product quality. , the effect of reducing labor intensity

Inactive Publication Date: 2014-11-19
武利 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the extraction process of cyclic adenosine monophosphate produced by fermentation is mainly ion exchange and adsorption chromatography. The biggest disadvantage of this method is the large amount of waste water, which is also a common problem in all

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The preparation of embodiment 1 fermented liquid

[0037] The fermented liquid used in the present invention is Arthrobacter fermented liquid, among which, the fermented liquid of Arthrobacter crystallopoietes, Arthrobacter oxidans, Arthrobacter CCTCC NO: M2013431, Arthrobacter sp.A302 is preferred. This embodiment provides a method for preparing the fermentation broth of the above-mentioned strains.

[0038] The concrete composition of medium used in this embodiment is as follows:

[0039] Incline medium (g / L): glucose 10, peptone 10, yeast extract 10, beef extract 10, NaCl3, agar 20. pH 6.8.

[0040] Seed medium (g / L): glucose 10, peptone 10, yeast extract 10, beef extract 10, NaCl3. pH 6.8.

[0041] Fermentation medium (g / L): glucose 30, K 2 HPO 4 15, KH 2 PO 4 5. MgSO 4 0.1, urea 10, biotin 0.005, CoCl 2 0.005, NaCl0.4, hypoxanthine 5, pH 7.0.

[0042] The fermentation conditions are as follows:

[0043] The -80°C refrigerator-preserved bacteria were ino...

Embodiment 2

[0045] This embodiment provides a method for extracting adenosine phosphate by ion exchange and resin chromatography, the steps are as follows:

[0046] 1) Take 1 L of the fermented liquid prepared in Example 1, the fermentation unit is 3 g / L, centrifuge at 5000 rpm for 5 min, collect the supernatant, and adjust the pH to 2.0 with concentrated hydrochloric acid.

[0047] 2) The resin used for ion exchange separation is 001×7 type cation exchange resin, the volume of the resin column is 200mL, and it is treated with 4% hydrochloric acid to form hydrogen form. The flow rate of the upper column is 0.3 column volume per hour. After the upper column, use 2 times the bed volume of purified water to press the material. After the pressing is completed, use 0.1N sodium hydroxide to elute. The elution flow rate is the same as above, and the eluent is collected 450mL .

[0048] 3) Resin chromatography The chromatographic resin is macroporous adsorption resin HP-20, and the column volume...

Embodiment 3

[0053] 1) Take 1 L of the fermentation broth prepared in Example 1, the fermentation unit is 4 g / L, centrifuge at 5000 rpm for 5 min, collect the supernatant, and adjust the pH to 3.0 with concentrated hydrochloric acid.

[0054] 2) The resin used for ion exchange separation is 001×7 type cation exchange resin, the volume of the resin column is 200mL, and it is treated with 4% hydrochloric acid to form hydrogen form. The flow rate of the upper column is 0.3 column volume per hour. After the upper column, use 2 times the bed volume of purified water to press the material. After the pressing is completed, use 0.1N sodium hydroxide to elute. The elution flow rate is the same as above, and the eluent is collected 450mL .

[0055] 3) Resin chromatography The chromatographic resin is macroporous adsorption resin HP-20, and the column volume of the resin is 300mL. The eluate collected after separation is added to the resin, and the sample loading flow rate is 0.3 column volume per ho...

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Abstract

The invention discloses a method for extracting cyclic adenosine monophosphate and application of cyclic adenosine monophosphate, and belongs to the technical field of extraction of biological active substances. The method provided by the invention comprises the following steps: adjusting the pH of a fermentation liquor by using an alkaline liquor; performing centrifugal separation to obtain a supernatant; concentrating the supernatant, and performing first discoloration; fractionally precipitating a discolored solution by using an organic solvent; dissolving an obtained precipitate, and performing second discoloration; performing micro porous filtration, and concentrating a second discolored solution; adding the organic solvent for precipitating the concentrated second discolored solution; and performing centrifugal separation, and drying the precipitate in vacuum to obtain a finished product. By adopting the method provided by the invention, the conventional ionic exchange and resin chromatography processes can be replaced, and the waste water amount can be lowered by 82.5 percent; meanwhile, the production period is shortened greatly, and the production time is shortened by 36 percent; and the prepared cyclic adenosine monophosphate is stable in quality, can be over 97 percent in purity, and can be widely applied to the fields of medicine, foods, feeds and the like.

Description

technical field [0001] The invention relates to a method and application for extracting cyclic adenosine monophosphate, and belongs to the technical field of microbial fermentation product extraction. Background technique [0002] Cyclic adenosine monophosphate is a second messenger substance involved in the regulation of cell function. It has a wide range of effects, which can increase myocardial contractility, cause blood pressure to increase, and increase cardiac output. It can relax smooth muscle, dilate coronary vessels, improve liver function, promote nerve regeneration, inhibit the division of epithelial cells in the outer layer of the skin and transform abnormal cells, promote the activity of respiratory chain oxidase, and improve myocardial hypoxia. [0003] At present, the main production method of cyclic adenosine monophosphate is chemical synthesis, which has the disadvantages of low yield, high cost, and high toxicity and side effects; in order to overcome these...

Claims

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Application Information

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IPC IPC(8): C07H19/213C07H1/06
Inventor 刘琬一王智敏
Owner 武利
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