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Method for producing adenosincoenzyme vitamin B12 by enzyme catalysis

A technology of adenosine coenzyme and vitamin, applied in the direction of fermentation, etc., can solve the problems of long production cycle, complicated process, high cost, etc., and achieve the effect of simple catalytic conditions, simple production process and easy purification

Inactive Publication Date: 2008-11-05
吉林省奇健生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The invention provides a method for producing adenosine coenzyme vitamin B12 by enzymatic catalysis, so as to solve the problems of complex process, long production cycle, high cost and heavy pollution in the current production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Enzyme Catalyzed Synthesis of Adenosine Coenzyme Vitamin B12 (1)

[0025] Take a one-liter reaction system as an example:

[0026] 1. Add reagents

[0027] Add the following prepared reagents to the reaction kettle: 50 ml of 1mol / l Tris-HCl buffer solution; 0.1mol / l CoCl 2 5ml; 0.1mol / l MnCl 2 10 ml; 0.5mol / l ATP 10 ml; 0.5mol / l hydroxycobalt vitamin B12 10 ml; deionized water to 1 liter; add Potasium Borohydride (KBH 4 )10 grams.

[0028] 2. Reaction temperature and time

[0029] Set the reaction temperature at 30±5°C, add 100 IU ATP:Vitamin B12 adenosyltransferase after 10 minutes, let the reaction time last for 2-3 hours, monitor the reaction progress with high performance liquid chromatography and UV / VIS spectrophotometer until The reaction is complete. Add hydrochloric acid to the above-mentioned mixed solution that finished the reaction, the final concentration is 0.1mol / l, centrifuge at 15000rpm for 10 minutes, discard the precipitate, and colle...

Embodiment 2

[0030] Example 2 Enzyme Catalyzed Synthesis of Adenosine Coenzyme Vitamin B12 (2)

[0031] Take a one-liter reaction system as an example:

[0032] 3. Add reagents

[0033] Add the following prepared reagents to the reaction kettle: 50 ml of 1mol / l Tris-HCl buffer solution; 0.1mol / l CoCl 2 5ml; 0.1mol / l MnCl 2 10 ml; 0.5mol / l ATP 20 ml; 0.5 mol / l hydroxycobalt vitamin B12 20 ml; deionized water to 1 liter; add Sodium Borohydride (NaBH 4 ) 15 grams.

[0034] 4. Reaction temperature and time

[0035] Set the reaction temperature to 30±5°C, add 200 IU ATP:Vitamin B12 adenosyltransferase after 10 minutes, and let the reaction time continue for 2-3 hours until the reaction is complete. Add hydrochloric acid to the above-mentioned mixed solution that finished the reaction, the final concentration is 0.1mol / l, centrifuge at 15000rpm for 10 minutes, discard the precipitate, and collect the supernatant for later use.

Embodiment 3

[0036]Example 3 Enzyme Catalyzed Synthesis of Adenosine Coenzyme Vitamin B12 (3)

[0037] Take a one-liter reaction system as an example:

[0038] 5. Add reagents

[0039] Add the following prepared reagents to the reaction kettle: 50 ml of 1mol / l Tris-HCl buffer solution; 0.1mol / l CoCl 2 10ml; 0.1mol / l MnCl 2 20ml; 0.5mol / l ATP 30ml; 0.5mol / l hydroxycobalt vitamin B12 30ml; deionized water to 1 liter; add Sodium Cyanoborohydride (NaBH 3 CN) 20 grams.

[0040] 6. Reaction temperature and time

[0041] Set the reaction temperature to 30±5°C, add 250 IU ATP:Vitamin B12 adenosyltransferase after 10 minutes, and let the reaction time continue for 2-3 hours until the reaction is complete. Add hydrochloric acid to the above mixed solution after the reaction is completed, the final concentration is 0.1mol / L, centrifuge at 15000rpm for 10 minutes, discard the precipitate, and collect the supernatant for later use.

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Abstract

This invention relates to a method for producing adenosine coenzyme vitamin B12 by enzyme catalyzation which utilizes the triphosphate adenosine: vitamin B12 adenosine transferring enzyme to produce the adenosine coenzyme vitamin B12, in which, the final concentration used by hydroxocobalt vitamin B12 is 0.001-0.05mol / l.

Description

technical field [0001] The invention relates to a production method of adenosine coenzyme vitamin B12, in particular, it refers to the production by utilizing biocatalysis engineering and medicinal chemical preparation related technologies. Background technique [0002] Adenosine coenzyme vitamin B12 (hereinafter referred to as adenosine coenzyme B12) is a new vitamin B12 preparation made by replacing the cyano group of ordinary vitamin B12 with 5′-deoxyadenosine nucleoside. It is the activated form of vitamin B12 in the body and has anti-anemia , antihistamine, liver protection, neuroprotection, protein assimilation, growth promotion and other physiological effects. Compared with common vitamin B12, adenosine coenzyme B12 has the advantages of low toxicity, direct utilization by the body, high affinity to tissues, high utilization rate in the body, and strong activity. Clinically, it is mainly used for the treatment of pernicious anemia and other megaloblastic anemia and o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/42
Inventor 陈大伟
Owner 吉林省奇健生物技术有限公司
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