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Production process for improving fermentative strength of 2-keto-L-gulonic acid

A production process, the technology of gulong acid, applied in the direction of fermentation, microorganism-based methods, microorganisms, etc., can solve the problems of restricting production efficiency, long cycle, high energy consumption, etc., and achieve the effect of overcoming long production cycle, reducing pollution, and simplifying the process

Inactive Publication Date: 2012-07-18
DSM JIANGSHAN PHARMACEUTICAL (JIANGSU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the foreign "Rayleigh method", the "two-step fermentation method" of vitamin C in China simplifies the process, reduces the cost and reduces the pollution. Physiological relationship, in the actual production, there are still technical problems that restrict production efficiency such as long cycle time and high energy consumption. Therefore, it is very important to improve the original fermentation production process and improve the production efficiency of 2-keto-L-gulonic acid fermentation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Control group: 1 liter of cultured common keto-based cologne and Bacillus megaterium mixed with 3.5 liters containing 2% sorbose, 1.1% corn steep liquor, 0.2% urea, 0.02% magnesium sulfate , 0.05% potassium dihydrogen phosphate fermentation medium, the fermentation vessel adopts a 10-liter automatic fermenter, the stirring speed is 400rpm, the aeration ratio is 1:1vvm, the initial pH of the medium is 6.7, and it is controlled by 25% sodium carbonate solution after 8 hours of cultivation The pH of the fermentation broth was 7.0, and sorbose solution with a concentration of 25% was added after 10 hours of cultivation, the final total sugar concentration of the fermentation was controlled to 10.2%, and the temperature was controlled at 30°C; the content of 2-keto-L-gulonic acid at the end of the fermentation broth was 99.26mg / ml, the fermentation period was 52 hours, and the acid production rate was 1.91mg / ml·h.

[0018] Experimental group: 1 liter of the cultured common ...

Embodiment 2

[0020] Control group: 10 liters of the cultured common keto-based cologne and Bacillus megaterium were mixed into 35 liters containing 2% sorbose, 1.1% corn steep liquor, 0.2% urea, and 0.02% magnesium sulfate , 0.05% potassium dihydrogen phosphate fermentation medium, the fermentation vessel adopts a 100-liter automatic fermenter, the stirring speed is 280rpm, the aeration ratio is 1: 0.8 (vvm), the initial pH of the medium is 6.7, and 25% carbonic acid is used for 8 hours of cultivation. Sodium solution controls the pH of the fermentation broth to 7.0, and starts to add sorbose solution with a concentration of 25% after 10 hours of cultivation, controls the final total sugar concentration of fermentation to 10.5%, and controls the temperature at 31°C; the end point of the fermentation broth is 2-keto-L-gulonic acid The content is 101.46mg / ml, the fermentation period is 50 hours, and the acid production rate is 2.03 mg / ml·h.

[0021] Experimental group: 10 liters of cultured ...

Embodiment 3

[0023] Control group: 6 m mixed cultured cultured common keto-cologronic acid bacteria and Bacillus megaterium 3 , access to 18m 3 In the fermentation medium containing 2% sorbose, 1.1% corn steep liquor, 0.2% urea, 0.02% magnesium sulfate, 0.05% potassium dihydrogen phosphate, the fermentation vessel adopts 50m 3 Standard fermentation tank, stirring speed 150rpm, aeration ratio 1: 0.4 (vvm), medium initial pH 6.7, 25% sodium carbonate solution was used to control the pH of the fermentation broth after 8 hours of cultivation, and the feed concentration was 25 after 10 hours of cultivation. % sorbose solution, control the final total sugar concentration of fermentation to 10.8%, and control the temperature at 30°C; the content of 2-keto-L-gulonic acid at the end of the fermentation liquid is 102.19 mg / ml, the fermentation period is 49 hours, and the acid production rate is 2.09 mg / ml h.

[0024] Experimental group: 6 m of cultured mixed bacteria liquid of common keto-cologron...

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Abstract

The invention discloses a production process for improving the fermentative strength of 2-keto-L-gulonic acid. The production process comprises the following steps of: controlling the temperature to be 28 to 33 DEG C during culture for 0 to 32 hours in the process of preparing the 2-keto-L-gulonic acid by fermenting mixed bacteria of Ketogulonigenium vulgare and Bacillus megaterium; culturing for 32 hours until the fermentation is finished, and controlling the temperature to be 34 to 38 DEG C; supplementing 0.2 to 0.5 percent of corn steep liquor during culture for 16 to 20 hours; and supplementing 0.1 to 0.35 percent of urea during culture for 28 to 32 hours. The physiological relationship of double bacteria in the fermentation of the2-keto-L-gulonic acid is adjusted through the research and development of stage-by-stage and fed-batch culture, so that environments and nutritional conditions are more suitable for the growth of mixed strain cells; therefore, the biosynthesis rate of a mixed strain is effectively improved; and strains are unchanged, and the process is easy to operate and popularize, low in cost, energy-saving and environment-friendly.

Description

Technical field: [0001] The invention relates to a preparation method of vitamin C, belongs to the technical field of microbial fermentation, and in particular relates to a new process for improving the fermentation production efficiency of 2-keto-L-gulonic acid. Background technique: [0002] 2-keto-L-gulonic acid (hereinafter referred to as 2-KLG) is an important component of vitamin C synthesis. At present, the main vitamin C manufacturers in China all adopt the "two-step fermentation" production process. The key step of this technology is the second step fermentation process: L-sorbose is produced in common ketogenic cologne (commonly known as small bacteria in the industry). ) and Bacillus megaterium (commonly known as large bacteria in the industry) are biologically oxidized to 2-KLG under the action of a mixed strain of bacteria. Compared with the foreign "Rayleigh method", the "two-step fermentation method" of vitamin C in China simplifies the process, reduces t...

Claims

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Application Information

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IPC IPC(8): C12P39/00C12P7/60C12R1/01C12R1/11
Inventor 黄建民杜尔凤肖江锋邵丽琴王志伟马华
Owner DSM JIANGSHAN PHARMACEUTICAL (JIANGSU) CO LTD
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