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Fermentation medium, bacterial strain and production method for fermentation production of adenosine cyclophosphate

A fermentation medium and cyclic adenosine monophosphate technology, applied in the field of bioengineering, can solve the problems of expensive raw materials, complicated process, and many by-products, and achieve the effect of low raw material cost, simple operation, and many by-products

Active Publication Date: 2014-02-12
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently used synthetic methods are all chemical synthesis methods, which have the disadvantages of complex process, expensive and difficult to obtain raw materials, many by-products, low yield, high cost and small output, which will cause environmental pollution and be unfavorable for industrial production.
At present, there is no literature report on the production of cyclic adenosine monophosphate by biotransformation

Method used

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  • Fermentation medium, bacterial strain and production method for fermentation production of adenosine cyclophosphate
  • Fermentation medium, bacterial strain and production method for fermentation production of adenosine cyclophosphate
  • Fermentation medium, bacterial strain and production method for fermentation production of adenosine cyclophosphate

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Embodiment 1

[0022] This embodiment is a screening method for bacterial strains that ferment and produce cyclic adenosine monophosphate, comprising the following steps:

[0023] Soil samples collected from a soil rich in organic matter are inoculated in an enrichment medium with hypoxanthine as a substrate for enrichment and isolation culture. Focusing on the appearance of the colony, observe the shape of the bacteria with a microscope, and obtain a number of Arthrobacter strains. After multiple purifications, they were respectively inserted into the slope, and stored in the refrigerator at 4°C for later use. The five purified strains were selected and named A.sp01, A.sp02, A.sp03, A.sp04, A.sp5 respectively.

[0024] A.sp01, A.sp02, A.sp03, A.sp04, A.sp5 strains were respectively inoculated in 100mL microbial enrichment medium for shaking culture at 42°C, and the conversion of hypoxanthine was measured every day. After complete conversion, the conversion time was recorded, and A.sp01 had...

Embodiment 2

[0052] The fermentation medium for producing cyclic adenosine monophosphate in this example includes the following mass fractions: glucose 5%, peptone 0.5%, potassium dihydrogen phosphate 1%, dipotassium hydrogen phosphate 1%, magnesium sulfate 0.3%, urea 1%, purine base 0.3%; pH natural.

[0053] The method for fermentatively producing cyclic adenosine monophosphate of the present embodiment comprises the following steps:

[0054] 1) Inoculate the A.sp01 strain on the prepared slant medium, and incubate at a constant temperature of 30°C for 48 hours until a large number of colonies grow;

[0055] 2) Pick colonies and inoculate them in the seed medium for 3 days with shaking at 28°C and 160rpm;

[0056] 3) Inoculate the seed liquid in step 2) into the fermentation medium at an inoculum amount of 5% for fermentation and culture, and ferment and cultivate for 60 hours at 28°C and 60% dissolved oxygen.

Embodiment 3

[0058] The fermentation medium for producing cyclic adenosine monophosphate in this example includes the following mass fractions: glucose 5%, peptone 0.5%, potassium dihydrogen phosphate 1%, dipotassium hydrogen phosphate 1%, magnesium sulfate 0.3%, urea 1%, purine base 0.3%; pH natural.

[0059] The method for fermentatively producing cyclic adenosine monophosphate of the present embodiment comprises the following steps:

[0060] 1) Inoculate the A.sp01 strain on the prepared slant medium, and incubate at a constant temperature of 33°C for 42 hours until a large number of colonies grow;

[0061] 2) Pick colonies and inoculate them in the seed medium for 1 day with shaking at 37°C and 160rpm;

[0062] 3) Inoculate the seed solution in step 2) into the fermentation medium at an inoculum amount of 5% for fermentation and culture, and ferment and cultivate for 65 hours at 32°C and 62% dissolved oxygen.

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Abstract

The invention discloses a fermentation medium, a bacterial strain and a production method for fermentation production of adenosine cyclophosphate. The medium comprises the following components in percentage by mass: 5-15% of glucose, 0.5-1% of peptone, 1-6% of monopotassium phosphate, 1-6% of dipotassium phosphate, 0.3-0.6% of magnesium sulfate, 1-6% of urea, and 0.3-0.6% of purine base. The bacterial strain is Arthrobacter sp.A.sp01; the preservation number is CCTCC NO:M 2013431; the preservation data is September 17, 2013; the preservation authority is the China center for type culture collection; the preservation address is Wuhan university in China. The fermentation medium disclosed by the invention has the advantages of being simple to operate, low in raw material cost, safe and environment-friendly, and can be applied to industrial production. An experiment proves that the content of the adenosine cyclophosphate obtained by the production method disclosed by the invention is at least more than 7.8g / L, and can be up to 9.16g / L. A new route is opened for fermentation production of the adenosine cyclophosphate by employing microorganisms.

Description

technical field [0001] The invention relates to a fermentation medium for fermenting and producing cyclic adenosine monophosphate, and also relates to a bacterial strain and a production method using the medium to produce cyclic adenosine monophosphate, belonging to the technical field of bioengineering. Background technique [0002] Cyclic adenosine monophosphate is adenosine-3′,5′-cyclic phosphate, referred to as cAMP, which is a substance that exists widely in the human body and has important physiological activities. Adenosine is catalyzed by adenosine activating enzyme. As a second messenger substance involved in the regulation of cell functions, it plays a role in hormone regulation of physiological functions and substance metabolism in cells, and plays an important role in sugar, fat metabolism, synthesis and regulation of nucleic acid and protein. [0003] Cyclic adenosine monophosphate is a protease activator, a nucleotide derivative, and an important pharmaceutica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/32C12R1/06
Inventor 常景玲王洪立李东霄张志宏张勉李畅孙寅玮
Owner HENAN INST OF SCI & TECH
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