Method for improving production capacity of 2-KGA (2-keto-L-gulonic acid) through enhancement of Ketogulonogeniumvulgarum carbon metabolism level

A technology of keto-gulonic acid bacteria and gulon acid, which is applied in the field of transforming vitamin C fermentation technology, and can solve problems such as unseen

Inactive Publication Date: 2015-02-18
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Before the disclosure of the present invention, there were no reports or patents on the application

Method used

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  • Method for improving production capacity of 2-KGA (2-keto-L-gulonic acid) through enhancement of Ketogulonogeniumvulgarum carbon metabolism level
  • Method for improving production capacity of 2-KGA (2-keto-L-gulonic acid) through enhancement of Ketogulonogeniumvulgarum carbon metabolism level
  • Method for improving production capacity of 2-KGA (2-keto-L-gulonic acid) through enhancement of Ketogulonogeniumvulgarum carbon metabolism level

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Implementation Example 1: Investigate the impact of adding two-carbon and six-carbon units on the ability to produce 2-KGA in conventional seed solution under single factor conditions

[0027] The concentrations of the above-mentioned two-carbon and six-carbon unit carbon sources added to each conventional seed medium were 0.02%, 0.2%, and 2% in sequence.

[0028] Conventional seed medium is: 2% L-sorbose, 0.5% corn steep liquor, 0.2% glucose, 0.1% urea, 0.2% calcium carbonate, pH6.7-7.0.

[0029] The conventional fermentation medium is: 8% L-sorbose, 1.5% corn steep liquor, 0.1% potassium dihydrogen phosphate, 0.1% urea, 0.01% magnesium sulfate, 0.5% calcium carbonate, pH6.7-7.0.

[0030] Culture conditions: the slant of the mixed bacteria culture was inoculated in common seed liquid medium with different two-carbon or six-carbon units, 30°C, 220 rpm, shake flask culture for 20 hours to make seed liquid, 10% was inoculated in ordinary fermentation In the culture mediu...

Embodiment 2

[0035] Implementation Example 2: Investigate the impact of adding two-carbon and six-carbon units on the ability to produce 2-KGA in conventional fermentation broth under single factor conditions

[0036] The above-mentioned two-carbon and six-carbon unit carbon sources were respectively added to each conventional fermentation medium at a concentration of 0.025%, 0.25% and 2.5%.

[0037] Conventional seed medium: see Example 1.

[0038] Conventional fermentation medium is: referring to example 1.

[0039] Culture conditions: see Example 1. The experimental results are shown in Table 2.

[0040] Table 2. Under single factor conditions, the effects of adding 0.025%, 0.25% and 2.5% two-carbon and six-carbon unit carbon sources on fermentation in the fermentation broth

[0041]

[0042][Note] The bold font is the effect of the optimum concentration of active substances on fermentation.

[0043] It can be seen from Table 2 that the above-mentioned 6 kinds of carbon sources al...

Embodiment 3

[0044] Implementation example 3: The effect of adding the optimized combination of two carbon and six carbon units to the seed liquid or fermentation liquid on the ability to produce 2-KGA

[0045] Conventional seed medium: see Example 1.

[0046] Conventional fermentation medium is: referring to example 1.

[0047] Culture conditions: see Example 1.

[0048] Using the orthogonal analysis method to add different combinations of two-carbon and six-carbon units in the seed solution, and taking the fermentation period of the fermentation broth as the result, investigate the effect of adding different combinations of two-carbon and six-carbon units in the seed solution on the final fermentation cycle. The experimental results are shown in table 3.

[0049] Table 3.L 32 Orthogonal analysis results of (4^6) 4 levels and 6 factors seed solution adding 2-carbon and 6-carbon unit combinations.

[0050]

[0051]

[0052] [Note] The bold font is the effect of the optimum concen...

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Abstract

The invention discloses a method which enhances glycometabolism efficiency of Ketogulonogeniumvulgarum, increases the number of bacteria, greatly shortens the incubation period of 2-KGA (2-keto-L-gulonic acid) and improves the production efficiency by adding carbon sources including two-carbon units such as ethyl alcohol, acetaldehyde and acetic acid as well as six-carbon units such as D-sorbitol, D-mannitol and D-mannose which facilitates utilization of Ketogulonogeniumvulgarum in a mixed fermentation process of a vitamin C two-step fermentation method according to research results of Ketogulonogeniumvulgarum metabolic network analysis. In a 5L fermentation tank, the period can be shortened by 14.04%, and the yield is increased by 3.40 %. According to another application, L-sorbose is taken as a carbon source by reducing Ketogulonogeniumvulgarum, so that the efficiency for converting L-sorbose into 2-KGA is also improved. According to the method, the production process is advanced, the raw material cost is low, and the method is suitable for large-scale industrial production and is improvement and promotion of vitamin C preparation technology with a biological fermentation method in about forty years of China; and besides, harmful substance emission in a production process is prevented, pollution and public hazard to the ambient environment are avoided, and environment protection requirements are met.

Description

Technical field: [0001] The invention belongs to the field of microbial fermentation, and relates to a method for improving the 2-KGA (2-keto-L-gulonic acid) production capacity of ketoglobulina bacteria by selecting and optimizing the composition of microbial carbon sources, and transforming vitamin C fermentation process. Background technique: [0002] Vitamin C (VC) is an essential vitamin for the human body. It has a wide range of physiological functions in anti-oxidation and maintaining metabolic balance. It has important uses in the pharmaceutical industry, food industry, feed industry and cosmetics industry. It has a wide range of applications and a huge market. [0003] At present, more than 90% of the world's VC production uses the "two-step fermentation method" invented by my country. The first step of fermentation is to oxidize D-sorbitol to L-sorbose under the action of Gluconobacter oxidans. The second step is to further oxidize L-sorbose to 2-keto-L-gulonic ...

Claims

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Application Information

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IPC IPC(8): C12P39/00C12P7/60C12R1/11C12R1/01
Inventor 张怡轩付阳于哲张盛李野韩立涛张海宏
Owner SHENYANG PHARMA UNIVERSITY
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