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Method for producing guanosine by fermentation with bacillus

A technology of bacillus and guanosine, which is applied in the field of guanosine production, can solve the problems of no patent reports, etc., and achieve the effects of removing metabolic regulation, low cost of use, and simple operation

Active Publication Date: 2013-07-10
JIANGSU ANHUI BIO TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are domestic literature reports using multi-scale theories and methods of biochemical processes to study the fermentation process of guanosine. By adding metabolic regulators to the fermentation medium, combined with the adjustment of fermentation parameters, the accumulation of amino acids, organic acids, and ammonium ions can be inhibited, and the accumulation of ammonium ions can be prevented. The migration of guanosine metabolic flow in the later stage of fermentation allows more carbon sources to flow to the guanosine production pathway and increases the fermentation yield of guanosine, but there is no patent report on this method so far

Method used

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  • Method for producing guanosine by fermentation with bacillus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) Cell activation

[0038] Streak inoculation of Bacillus subtilis CGMCC No.4587 on the slant of nutrient broth agar test tube, then transfer to the slant of agar eggplant bottle to cultivate and mature, ready for transplantation; culture medium and agar eggplant for nutrient broth agar test tube slant culture The culture medium for bottle slant culture is: glucose 5g / L, peptone 10g / L, yeast extract 10g / L, NaCl 5g / L, agar 20g / L, pH 7.0-7.2, culture temperature uniform 32℃, culture time uniform for 24-48 hours.

[0039] (2) Seed cultivation

[0040] Bacillus subtilis JSIM-G518 slant refrigerated strains were streaked and inoculated on the slant of nutrient broth agar test tube. The slant medium consisted of: glucose 5g / L, peptone 10g / L, yeast extract 10g / L, NaCl 5g / L L. Agar 20g / L, pH 7.2, cultured at 32°C for 48 hours, it is mature.

[0041] On the clean bench in the sterile room, pick a ring of activated test tube slant bacteria and inoculate it into the sterilize...

Embodiment 2

[0047] Step (1) is with embodiment 1.

[0048] Step (2) Seed culture

[0049] Bacillus subtilis JSIM-G518 slant refrigerated strains were streaked and inoculated on the slant of nutrient broth agar test tube. The slant medium consisted of: glucose 5g / L, peptone 10g / L, yeast extract 10g / L, NaCl 5g / L L. Agar 20g / L, pH 7.2, cultivated at 32°C for 48 hours. Then transfer to the slant of the agar eggplant bottle (the composition of the medium is the same as that of the test tube slant), culture at 32° C. for 48 hours, and mature.

[0050] Prepare a sterile triangular flask with glass beads inside, scrape and wash the mature fungal slime on the slope of the eggplant bottle with sterile water, put it into the triangular flask, oscillate and disperse the fungal slime to obtain a uniform bacterial suspension; press the bacterial suspension to 10% of the inoculum was inserted into a 100-liter seed tank, and the filling factor of the seed tank was 70% (V / V). The composition of the see...

Embodiment 3

[0060] 1. Collect 70 liters of guanosine fermentation liquid, send it into the storage tank, heat the jacket to about 95°C, and keep it warm for 15 minutes; filter it with a plate and frame filter press while heating, and the filtration pressure is 0.40Mpa. The bacteria are used as feed protein, and the filtrate is sent to the storage tank;

[0061] 2. Add 0.2% volume of natural flocculant ZTC-II (or one of chitosan, chitin, and sodium alginate) to the filtrate (prepared to a solution with a weight concentration of 1%), and keep the jacket at 32°C , flocculation and sedimentation for 12 hours to remove protein, take the supernatant and send it to the storage tank.

[0062] 3. The supernatant was treated with a ceramic membrane, and the membrane filtration temperature was 75°C; the filtrate was concentrated to 1 / 5 volume under reduced pressure and vacuum at 60°C.

[0063] 4. After adjusting the pH of the concentrated solution to 7.0, send it to a crystallization tank, stir, an...

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Abstract

The invention discloses a method for producing guanosine by fermentation with bacillus, which sequentially comprises the following steps: bacillus activation, seed culture, fermentation culture, guanosine extraction and refinement, and the like. The strain is Bacillus subtilis CGMCC No.4587. The invention increases the fermentation yield and conversion rate of guanosine.

Description

technical field [0001] The present invention relates to a method for producing guanosine. Background technique [0002] Guanosine is also known as guanosine nucleoside, the chemical name is 9-β-D-ribofuranosylguanine, the English name is Guanosine, and the molecular formula is C 10 h 13 N 5 o 5 , molecular weight 283.24, guanosine is a white crystalline powder, odorless, tasteless; the UV absorption peak of the aqueous solution is at 256 μm, and the molar extinction coefficient is 12.2×10 3 ; Dissolve 1 gram in 1320ml water at 18°C; Dissolve 1 gram in 33ml water in a boiling water bath; Soluble in acid and alkali, insoluble in organic solvents; Melting point 237-240°C. [0003] Guanosine belongs to nucleoside small molecule compounds, and is mainly used in industry as a key intermediate in the synthesis of antiviral and antitumor drugs, and as a raw material for large-scale synthesis of food additive 5'-guanylic acid (GMP). [0004] 5'-guanylic acid is a strong taste ag...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/40C12R1/125
Inventor 郑惠华匡群孙梅张一平华洵璐何寅琛叶进诸建国
Owner JIANGSU ANHUI BIO TECH
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