Method for fermentation production of cyclic adenosine monophosphate by controlling dissolved oxygen content at two phases

A cyclic adenosine monophosphate and dissolved oxygen technology, which is applied in the field of microbial fermentation, can solve problems such as limited production scale, and achieve the effects of less environmental pollution, mild reaction conditions and less by-products

Active Publication Date: 2013-01-30
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

But the production scale of this method is limited (only reaching the gra

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; Autoclaved at 121°C for 15 minutes.

[0036] Add the culture medium to the fermenter (BioFlo 110 7.5L NEW BRUNSWICK SCIENTIFIC), the liquid volume is 3L, after sterilization, inoculate with 10% inoculum, the temperature is 30°C, the ventilation rate is 1.5vvm, the rotation speed is 300r / min, and the dissolved oxygen is calibrated percentage point. The dissolved oxygen parameter is related to the speed of the impeller, and with appropriate ventilation, the dissolved oxygen is controlled to be 30% in the first stage of fermentation (about 0~18h), and the dissolved oxygen is controlled to be 10% in the second stage (about 18~60h). %. The pH was controlled to 7.0 by feeding ammonia water with a mass concentration of 20%.

[0037] After 60 hours of fermentation, HPLC detection showed that the cAMP content...

Embodiment 2

[0039] Fermentation medium (g / L): molasses 50, dipotassium hydrogen phosphate 10, potassium dihydrogen phosphate 10, magnesium sulfate 10, peptone 4, biotin 0.003, adenosine 6, sterilized by high-pressure steam at 121°C for 15 minutes.

[0040] Add the culture medium to the fermenter (BioFlo 1107.5L NEW BRUNSWICK SCIENTIFIC), the liquid volume is 3.5L, after sterilization, inoculate with 8% inoculum, the temperature is 32°C, the ventilation rate is 1.2vvm, the rotation speed is 350r / min, and the dissolved oxygen is calibrated percentage point. The dissolved oxygen parameter is related to the speed of the impeller, and with appropriate ventilation, the dissolved oxygen is controlled at 30% in the first stage of fermentation (about 0~18h), and the dissolved oxygen is controlled at 8% in the second stage (about 18~60h). %. The pH was controlled to 7.2 by feeding sodium hydroxide with a mass concentration of 20%.

[0041] After 54 hours of fermentation, HPLC detection showed tha...

Embodiment 3

[0043] Fermentation medium (g / L): 50 sucrose, 10 dipotassium hydrogen phosphate, 10 potassium dihydrogen phosphate, 10 magnesium sulfate, 4 peptone, 0.003 biotin, 6 adenosine triphosphate, sterilized by high-pressure steam at 121°C for 15 minutes.

[0044] Add the culture medium into the fermenter (BioFlo 110 7.5L NEW BRUNSWICK SCIENTIFIC), the liquid volume is 4L, after sterilization, then inoculate with 5% inoculum, the temperature is 28°C, the ventilation rate is 1.0vvm, the rotation speed is 400r / min, and the calibration solution oxygen percentage. The dissolved oxygen parameter is related to the impeller speed, and with appropriate ventilation, the dissolved oxygen is controlled at 25% in the first stage of fermentation (about 0~18h), and the dissolved oxygen is controlled at 5% in the second stage (about 18~60h). %. The pH was controlled to 7.5 by feeding potassium hydroxide with a mass concentration of 20%.

[0045] After 60 hours of fermentation, HPLC detection showe...

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Abstract

The invention provides a method for fermentation production of cyclic adenosine monophosphate by controlling the dissolved oxygen content at two phases. The method adopts the two-phase dissolved oxygen content control manner to carry out batch fermentation in order to produce the cyclic adenosine monophosphate, fermentation strains are Arthrobacter sp having a preservation number of CGMCC No.3584, the dissolved oxygen in a thallus growth phase is 20-30%, and the dissolved oxygen is 5-10% after the thallus amount stabilizes. The method provided by the invention has the advantages of substantial improvement of the output of the cyclic adenosine monophosphate, no need of the addition of extra equipment in the fermentation process, saving of the electric energy for stirring and air supply in the later fermentation phase, and suitableness for industrialized production.

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 two-stage dissolved oxygen controlled fermentation. Background technique [0002] Cyclic adenosine monophosphate (cAMP for short) is a derivative of nucleotide, which plays an important regulatory role in the metabolism of sugar and fat, and the synthesis of nucleic acid and protein. Clinically, cyclic adenosine monophosphate can inhibit cancer cells under certain conditions, and the determination of cAMP metabolism levels in plasma and body tissues has certain reference value for early diagnosis of tumor patients, tumor monitoring and patient prognosis; cAMP can activate fat Decomposing enzymes, promoting fat decomposition and lipid metabolism, and preventing atherosclerosis (Niu Shuling, Liu Jingbo. 1996. The application effect of cyclic nucleotides. Jilin Animal Husbandry and Veterinary Medicine, 5: 42-4...

Claims

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Application Information

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IPC IPC(8): C12P19/32C12R1/06
Inventor 应汉杰陈晓春李磊柏建新陈勇谢婧婧吴菁岚
Owner NANJING UNIV OF TECH
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