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
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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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