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Method for producing cyclic adenosine monophosphate through middle feeding fermentation

A technology of cyclic adenosine monophosphate and cyclic phosphoric acid, applied in the field of microbial fermentation, can solve problems such as limited production scale, and achieve the effects of low production cost, stable high-yield properties, and low carbon source concentration

Active Publication Date: 2015-01-28
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the scale of production is limited (only reaches the gram scale), and there is still a long way to go from real industrialization.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035]Fermentation medium (g / L): glucose 50, dipotassium hydrogen phosphate 10, potassium dihydrogen phosphate 10, magnesium sulfate 1, peptone 4, biotin 0.003, hypoxanthine 6, pH 6.0, autoclaved at 121°C for 15 minutes .

[0036] Arthrobacter A302 on the slant test tube was connected with 2 loops to the seed medium, cultivated for 18 hours at 30° C. on a shaking table at 200 r / min, and obtained the seed solution in the exponential phase.

[0037] Insert the seed liquid into a fermenter containing 3L of fermented liquid according to the inoculation amount of 8% (v / v) of the fermentation volume. The fermentation temperature is 28°C, the ventilation rate is 1.5vvm, and the stirring speed is 350r / min. , 44h, and 52h, add 600g / L glucose solution at a constant speed, once every 8h, 5 times, 40mL each time, and the amount of feeding is about 200ml.

[0038] After culturing for 72 hours, HPLC detection showed that the cAMP content of the product was 11.21g / L, and the production inte...

Embodiment 2

[0040] Fermentation medium (g / L): glucose 45, dipotassium hydrogen phosphate 10, potassium dihydrogen phosphate 5, magnesium sulfate 0.5, peptone 4, biotin 0.005, adenine 6, pH 7.0, autoclaved at 121°C for 15 minutes.

[0041] Other conditions were the same as the method in Example 1. Arthrobacter A302 on the slant test tube was connected to the seed medium with 2 rings, and cultured at 30° C. on a shaker at 200 r / min for 18 hours to obtain the seed liquid in the exponential phase.

[0042] Put the inoculum of 10% of the fermentation volume (v / v) of the seed liquid into a fermenter of 3.5L fermentation liquid, the fermentation temperature is 32°C, the ventilation rate is 1.2vvm, the stirring speed is 300r / min, between 20 and 50h Glucose solution with a concentration of 400g / L was fed every 3 hours, 10 times in total, 20ml each time, and the feeding volume was about 200ml.

[0043] After culturing for 72 hours, HPLC detection showed that the cAMP content of the product was 10.8...

Embodiment 3

[0045] Fermentation medium (g / L): molasses 50, disodium hydrogen phosphate 10, sodium dihydrogen phosphate 10, magnesium sulfate 1, urea 4, biotin 0.005, adenosine 6, pH 6.0, autoclaved at 121°C for 15 minutes.

[0046] Other conditions were the same as the method in Example 1. Arthrobacter A302 on the slant test tube was connected to the seed medium with 2 rings, and cultured at 30° C. on a shaker at 200 r / min for 18 hours to obtain the seed liquid in the exponential phase.

[0047] Insert the inoculum of the seed liquid into a 4L fermentation liquid fermenter according to the inoculum amount of 4% (v / v) of the fermentation volume, the fermentation temperature is 34°C, the ventilation rate is 1.5vvm, and the stirring speed is 350r / min, at 20h, 26h, 32h, Add 300g / L molasses at a constant speed at 38h, 44h, and 50h respectively, once every 6h, 6 times, 30mL each time. The flow rate is about 180ml.

[0048] After culturing for 66 hours, HPLC detection showed that the content of...

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Abstract

The invention provides a method for producing cyclic adenosine monophosphate through middle feeding fermentation. The method for producing cyclic adenosine monophosphate through middle feeding fermentation allows the cyclic adenosine monophosphate to be produced through batch fermentation in an intermittent uniform-speed feeding manner, fermentation strains are Arthrobacter sp having a preservation number of CGMCC No.3584, and a feeding carbon source is intermittently fed after the exponential phase of fermentation in a uniform speed manner each 3-10h 4-10 times. The method has the advantages of low concentration of the carbon source in the whole culture process, basic exhaustion of the carbon source after the fermentation ending, feedback inhibition reduction, and great increase of the output of the cyclic adenosine monophosphate.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a method for producing cyclic adenosine monophosphate by intermediate feeding fermentation. Background technique [0002] Cyclic adenosine monophosphate (cAMP for short) is one of the important contents of current molecular biology research and has the function of a second messenger. Basic medical research has proved that at least 40 kinds of diseases (including cancer, hypertension, coronary heart disease, myocardial infarction and cardiogenic shock, psoriasis, etc.) are related to the metabolism of cAMP (Hong D, Peng X R.2003.Role of the cAMP in immunological liver injury in mice: comparing LPS-induced model with LPS+BCG-induced model. Chinese Pharmacol Bull. 19: 940~943). In terms of pharmaceuticals, cyclic adenosine monophosphate can also be used as a drug intermediate to prepare dibutyryl cyclic adenosine monophosphate and cyclic adenosine monophosphate meg...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/40C12R1/06
Inventor 应汉杰李磊陈晓春柏建新陈勇吴菁岚谢婧婧
Owner NANJING TECH UNIV
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