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A kind of preparation method of cyclic adenosine monophosphate

A technology of cyclic adenosine monophosphate and tricarboxylic acid, applied in the direction of using bacteria, methods based on microorganisms, biochemical equipment and methods, etc., to achieve the effect of reducing the formation of organic acids

Inactive Publication Date: 2011-12-07
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield of the existing fermentation method for producing cyclic adenosine monophosphate still needs to be further improved

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Fermentation culture medium is in ordinary culture medium (based on the weight percent of culture medium, comprises glucose 5%, K 2 HPO 4 1%, KH 2 PO 4 1%, MgSO 4 1%, urea 0.5%, peptone 0.5%), add 0.1g NaF, 0.1g VB1 and 5g hypoxanthine per liter of medium.

[0026] Culture method: insert Arthrobacter A302 into the seed medium (based on the weight percentage of the medium, containing 1% glucose, 1% peptone, 0.5% yeast extract, 1% beef extract, and 0.3% NaCl), the initial pH is 7.0 , cultivated at 30° C. and 240 rpm for 18 hours. 10% of the inoculum was inserted into the fermentation medium in a 5L fermenter, the pH was controlled to be 7.0 with NaOH, the dissolved oxygen was controlled to be 30%, and fermentation was carried out at 400 rpm and 30° C. for 72 hours. When putting into tanks, the output of cyclic adenosine monophosphate is 10.2g / L, which is 3.25 times higher than the output (2.4g / L) when only using common culture medium.

Embodiment 2

[0028] The fermentation medium was added 6 g of hypoxanthine per liter of medium on the basis of ordinary medium (same as Example 1). The cultivation method is the same as in Example 1. When putting into tanks, the output of cyclic adenosine monophosphate is 4.6g / L, which is 0.92 times higher than the output (2.4g / L) when only using common culture medium.

Embodiment 3

[0030] The fermentation medium was added with 0.2 g of sodium fluoride per liter of medium on the basis of ordinary medium (same as in Example 1). The cultivation method is the same as in Example 1. When putting into tanks, the output of cyclic adenosine monophosphate is 3.2g / L, which is 0.33 times higher than the output (2.4g / L) when only using common culture medium.

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Abstract

The present invention provides a method of preparing cyclic adenosine monophosphate, which includes the use of Arthrobacter sp. as an engineering bacterium, and the addition of precursors, inhibitors of glycolytic pathway and / or activators of tricarboxylic acid cycle into a fermentation medium, so as to acquire the cyclic adenosine monophosphate by fermentation. Further provided is a use of precursors, inhibitors of glycolytic pathway and / or activators of tricarboxylic acid cycle for preparing cyclic adenosine monophosphate by Arthrobacter sp.. The present invention inhibits the excessive glycolysis, increases the flux of pentose phosphate pathway, and enhances the tricarboxylic acid cycle by the addition of precursors, inhibitors of glycolytic pathway and / or activators of tricarboxylic acid cycle, so as to redistribute rationally the metabolic flux of central metabolic pathways, make the carbon flux more efficiently direct to the target product - cyclic adenosine monophosphate, and synthesize cyclic adenosine monophosphate by using carbon source efficiently.

Description

technical field [0001] The invention relates to a preparation method of cyclic adenosine monophosphate, in particular to a method for obtaining cyclic adenosine monophosphate with a high yield by using Arthrobacter as a production bacterium, adding regulating substances to a fermentation medium, and obtaining cyclic adenosine monophosphate with high yield through fermentation. Background technique [0002] Cyclic adenosine monophosphate (cAMP) is an important substance with physiological activity that widely exists in the human body. As a second messenger in cells, it plays an important role in the regulation of metabolism and synthesis of sugar, fat, nucleic acid, protein, etc. . It can be used clinically to treat angina pectoris, myocardial infarction, myocarditis and cardiogenic shock; it also has a certain effect on improving symptoms such as palpitations, shortness of breath and chest tightness of rheumatic heart disease; it can improve the curative effect of acute leuk...

Claims

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Application Information

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IPC IPC(8): C12P19/32C12R1/06
CPCC12R1/06C12P19/32C12N1/205C12R2001/06
Inventor 应汉杰柏建新陈晓春熊健陈勇
Owner NANJING UNIV OF TECH
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