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Method for producing cyclic adenosine monophosphate through middle feeding fermentation

A technology of cyclic adenosine monophosphate and feed feeding, which is applied in the field of intermediate fed feed fermentation to produce cyclic adenosine monophosphate, can solve the problems of limited production scale and achieve the effects of low production cost, increased final concentration, and low environmental pollution

Active Publication Date: 2013-01-30
NANJING UNIV OF TECH
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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, 121 ℃ high pressure steam sterilization for 15min .

[0036] The Arthrobacter A302 on the inclined tube was connected to the seed medium by 2 loops at 30° C., shaking at 200 r / min, and cultivated for 18 h to obtain the seed liquid in the exponential phase.

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

[0038] After culturing for 72h, the cAMP content of the product was 11.21g / L, and the production intensity was 0...

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, high pressure steam sterilization at 121° C. for 15 min.

[0041] The other conditions were the same as those in Example 1. Arthrobacter A302 on the inclined tube was connected to the seed medium for 2 loops at 30° C., shaking at 200 r / min, and cultured for 18 h to obtain the seed liquid in the exponential phase.

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

[0043] After culturing for 72 h, the cAMP content of the product...

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, high pressure steam sterilization at 121° C. for 15 min.

[0046] The other conditions were the same as those in Example 1. Arthrobacter A302 on the inclined tube was connected to the seed medium for 2 loops at 30° C., shaking at 200 r / min, and cultured for 18 h to obtain the seed liquid in the exponential phase.

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

[0048] After culturing for 66 h, the cAMP content of the product was 8.83 g / L, and the prod...

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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, in particular to a method for producing cyclic adenosine monophosphate by intermediate feed fermentation. Background technique [0002] Cyclic adenosine monophosphate (cAMP) is one of the important contents of current molecular biology research and has the function of second messenger. Basic medical research has proved that at least 40 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-indudced model. Chinese Pharmacol Bull. 19: 940-943). In terms of pharmaceuticals, cyclic adenosine monophosphate can also be used as a pharmaceutical intermediate to prepare dibutyryl cyclic adenosine monophosphate and meglumine cyclomonophosphate, which can improve the lipid s...

Claims

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Application Information

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Patent Type & Authority Applications(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 UNIV OF TECH
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