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Process method for improving fermenting property of cAMP (cyclic adenosine monophosphate) with polyphosphate

A technology of polyphosphate and cyclic adenosine monophosphate is applied in the field of microbial fermentation process optimization to achieve the effect of improving fermentation strength and other indicators

Pending Publication Date: 2019-03-01
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the traditional process, the production is carried out by adding precursors to the initial medium and supplementing carbon sources midway, and adding NaF and other EMP pathway inhibitors to rationally regulate the metabolic flow, and energy supply becomes the main limiting factor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Fermentation medium (g / L): glucose 80, dipotassium hydrogen phosphate 10, potassium dihydrogen phosphate 10, magnesium sulfate 3, peptone 5, biotin 0.03, hypoxanthine 5, urea 10, pH 7.4, 121°C high-pressure steam Sterilize for 20 minutes.

[0027] Arthrobacter sp.CCTCC 2013431 on the slant test tube was connected to the seed medium in 3 loops, 30°C, shaker 220r / min, and cultivated for 24h to obtain the seed solution in the exponential phase.

[0028] The seed liquid is inserted into the fermentor containing 5L fermented liquid by the inoculum amount of 15% (v / v) of the fermentation volume, the fermentation temperature is 32°C, the ventilation rate is 1.0vvm, and the stirring speed is 500r / min.

[0029] Cultivate until the end of fermentation for 72 hours, and detect by HPLC, the cAMP content of the product is 3.32g / L, and the production intensity is 0.046g / (L·h).

Embodiment 2

[0031] The composition of the fermentation medium, the concentration of each component and the main fermentation conditions and steps are the same as in Example 1.

[0032] After the fermentation was carried out for 36 hours, a sufficient amount of sodium tripolyphosphate was added at one time to make the concentration of the substance in the fermentation broth reach 5 g / L.

[0033] Cultivate until the end of 60h fermentation, HPLC detection, product cAMP content is 3.74g / L, production intensity is 0.062g / (L·h).

Embodiment 3

[0035] The composition of the fermentation medium, the concentration of each component and the main fermentation conditions and steps are the same as in Example 1.

[0036] After 36 hours of fermentation, a certain amount of hypoxanthine solution was pulsed every 8 hours to increase the concentration of the substance in the fermentation broth by 1 g / L, and added 4 times.

[0037] The cAMP content of the product was 2.92g / L and the production intensity was 0.041g / (L·h) when the fermentation was completed at 72h.

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Abstract

The invention provides a process method for improving fermenting property of cAMP (cyclic adenosine monophosphate) with polyphosphate. According to the related method, cAMP is produced with a processof adding a precursor substance and polyphosphate in a batch fermentation coupled pulse manner, and a fermentation strain is Arthrobacter sp. A. sp01 with the collection number of CCTCC NO:M2013431. After batch fermentation reaches a logarithmic phase (20-40 h), 1-5 g / L of polyphosphate is added once, and 1-5 g / L of the precursor substance is supplemented every 4-15 h and added 3-6 times in total.By means of the process method, the yield of cAMP is increased to 2.18 times that of an original process, the production efficiency is 119.5% higher than that of the original process, and the fermentation performance is significantly improved. The method has the advantages that addition of polyphosphate ensures supply of energy in the whole fermentation process, the pulse type addition strategy can keep the concentration of the precursor substance in a fermentation broth at the level of 1-5 g / L, the inhibition phenomenon can be avoided, and product synthesis cannot be affected by lack of theprecursor, so that the fermentation performance of cAMP is greatly improved.

Description

technical field [0001] The invention belongs to the field of microbial fermentation process optimization, and specifically relates to a process method for improving the fermentation performance of cyclic adenosine monophosphate by using polyphosphate. Background technique [0002] Cyclic adenosine monophosphate (cAMP) is a physiologically active substance that widely exists in the human body. As a second messenger in cells, it plays an important role in regulating glucose metabolism, fat metabolism, nucleic acid and protein synthesis. The substance is widely used as medicine and feed additive in the treatment of cardiovascular diseases and in the production of livestock and poultry products, and has very important application value. [0003] At present, cAMP is mainly produced by chemical synthesis, the raw materials and reagents involved are expensive, and the production cost is relatively high. The cost per kilogram of cAMP is about 6,000 yuan, which seriously restricts th...

Claims

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Application Information

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IPC IPC(8): C12P19/32C12R1/06
CPCC12P19/32
Inventor 李志刚常景玲陈宝峰张中华李兰
Owner HENAN INST OF SCI & TECH
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