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Method for producing cyclic adenosine monophosphate by using whole cell biocatalysis

一种环磷酸腺苷、生物催化的技术，应用在生物催化领域，能够解决酶的活性损伤等问题，达到加速扩散、易再生、半衰期长的效果

Active Publication Date: 2010-08-18
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The thinking of the present invention is: for the defect that the activity of the enzyme is easily damaged in the enzymatic reaction, polyhydric organic reagents can be added to protect the activity of the enzyme

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Microbial strain medium: glucose 50g / L, urea 2.0g / L, potassium dihydrogen phosphate 5.0g / L, magnesium sulfate 0.5g / L, biotin 0.05×10 -3 g / L.

[0027] The inoculum amount of the microbial strain was 10%, cultured on a shaker at 120 rpm at 30° C. for 24 hours, and centrifuged at 8000 rpm for 10 minutes. Take the sludge and store it at -10°C for later use.

Embodiment 2

[0028] Example 2: Synthesis of cyclic adenosine monophosphate using hypoxanthine.

[0029] Take a 500mL beaker, prepare 4g / L hypoxanthine from 50g / L glucose, 100g Arthrobacter bacteria sludge, 8g / L potassium dihydrogen phosphate, 1g / L MgSO 4 , 1g / L NH 4 Cl, 4.5mL toluene, 0.5g / L mannitol, 0.5g / L xylitol, and 300mL of water, adjust the pH to 6.5 with sodium hydroxide, and stir at a low speed for 8 hours at 30°C. After the reaction , precipitated with 40% trichloroacetic acid, quantitatively analyzed cyclic adenosine monophosphate by HPLC, and the conversion solution contained cyclic adenosine monophosphate 4.5g / L.

Embodiment 3

[0030] Example 3: Synthesis of cyclic adenosine monophosphate using adenosine.

[0031] Take a 500ml beaker, and prepare 6g / L adenosine, 60g / L glucose, 120g air-dried microbacteria sludge, 10g / L potassium dihydrogen phosphate, 10g / L dipotassium hydrogen phosphate, 0.5g / L MgCl 2 , 2g / L NH 4 Cl, 1g / L glycerin, 1g / L mannitol, and 300mL reaction liquid, adjust pH to 7.0 with potassium hydroxide, stir at 33°C for 6h at low speed, after the reaction, precipitate with 40% trichloroacetic acid , Quantitative analysis of cyclic adenosine monophosphate was carried out by HPLC, and the conversion liquid contained cyclic adenosine monophosphate 3.5g / L.

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Abstract

The invention discloses a method for producing cyclic adenosine monophosphate by using whole cell biocatalysis, which is characterized in that the method comprises the following steps: using polyhydroxy organic reagents for protecting the activity of enzymes; using cyclic adenosine monophosphate precursors and phosphate anions as substrates; using glucose as energy suppliers; adding metal ions; and using permeable microbial bacterial strains for producing the cyclic adenosine monophosphate through whole cell biocatalysis. The invention uses the metal ion combination for regulating and controlling the metabolic flow rate for improving the energy self coupling efficiency, the organic reagents are added for protecting the thalli and the activity of enzymes, the permeable microbial bacterial strains are used for preparing the cyclic adenosine monophosphate, the synthesis time is shortened, and products can be accumulated outside the cells, so the cost of the subsequent separation can be saved, and the operation step can be simplified.

Description

technical field [0001] The invention belongs to the technical field of biocatalysis, and in particular relates to a method for producing cyclic adenosine monophosphate by utilizing whole cell biocatalysis. Background technique [0002] Cyclic adenosine monophosphate 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 sugar, fat metabolism, nucleic acid, protein synthesis and so on. It is clinically used to treat angina pectoris, myocardial infarction, myocarditis and cardiogenic shock; it 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 leukemia combined with chemotherapy, and can also be used for acute leukemia Induction remission; in addition, it also has certain curative effect on elderly chronic bronchitis, various hepatitis a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/32C12R1/06C12R1/01C12R1/13C12R1/19C12R1/125C12R1/645C12R1/15
CPCC12N1/20C12N11/16C12P19/32
Inventor 应汉杰柏建新宋贺陈晓春熊健苑巍陈勇
Owner NANJING TECH UNIV
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