Method for repairing metabolic defects of ketogulonigenium sp. cofactors and improving 2-KGA (2-keto-L- gulonic acid) producing capacity

A technology for cofactor and production capacity of uric acid bacteria, which is applied in the field of microbial fermentation to achieve the effects of improving electron transfer efficiency, increasing yield and improving bacterial growth.

Active 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

Simultaneously, before the disclosure of the present invention, there was no report on applying

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  • Method for repairing metabolic defects of ketogulonigenium sp. cofactors and improving 2-KGA (2-keto-L- gulonic acid) producing capacity
  • Method for repairing metabolic defects of ketogulonigenium sp. cofactors and improving 2-KGA (2-keto-L- gulonic acid) producing capacity
  • Method for repairing metabolic defects of ketogulonigenium sp. cofactors and improving 2-KGA (2-keto-L- gulonic acid) producing capacity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Implementation Example 1: Investigating the Effect of Adding Small Bacterial Metabolism Deficient Substances on the Production Capacity of 2-KGA in the Seed Solution under Single Factor Conditions

[0041] 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.

[0042] 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.

[0043] Culture conditions: the slant of the mixed bacteria culture is inoculated in the conventional seed liquid medium, 30°C, 220 rpm, shake flask culture for 20 hours to make the seed liquid, 10% is inoculated into the conventional fermentation culture with different metabolic defective substances of small bacteria added medium, 30°C, 220 rpm, shake flask culture until the substrate L-sorbose is less than 1mg / L and the fermentation ends.

[0044]...

Embodiment 2

[0049] Implementation Example 2: Investigating the Effect of Adding Small Bacterial Metabolism Deficient Substances on the Production Capacity of 2-KGA in Conventional Fermentation Liquid under Single Factor Conditions

[0050] The concentrations of the microbacterial metabolic defect substances added respectively in the conventional fermentation medium are shown in Table 2.

[0051] Conventional seed medium: see Example 1.

[0052] Conventional fermentation medium is: referring to example 1.

[0053] Culture conditions: see Example 1. The experimental results are shown in Table 2.

[0054] Table 2. The effect of adding microbacterial metabolic defect substances in the fermentation broth on fermentation under single factor conditions.

[0055]

[0056] [Note] The bold font is the effect of the optimum concentration of active substances on fermentation.

[0057] It can be seen from Table 2 that the above-mentioned 5 bacterial metabolic defect substances all promoted the g...

Embodiment 3

[0058] Implementation Example 3: The effect of adding the optimized small bacterial metabolic defect substance combination to the seed liquid or fermentation liquid on the ability to produce 2-KGA

[0059] Conventional seed medium: see Example 1.

[0060] Conventional fermentation medium is: referring to example 1.

[0061] Culture conditions: see Example 1.

[0062] The orthogonal analysis method was used to add different combinations of metabolic defect substances to the seed solution, and the fermentation period of the fermentation broth was used as the result to investigate the effect of adding different combinations of active substances in the seed solution on the final fermentation cycle. The experimental results are shown in Table 3.

[0063] Table 3.L 16 The results of (4^5)4-level 5-factor orthogonal analysis.

[0064]

[0065] [Note] The bold font is the effect of the optimum concentration of active substances on fermentation.

[0066] As can be seen from Tabl...

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Abstract

The invention discloses a method for improving the respiratory chain electron transport level of the ketogulonigenium sp. and increasing the thallus metabolic efficiency by adding a cofactor ubiquinone (CoQ10) and menaquinone (VK2) (in oxidized forms and reduced forms) as well as precursors including PHBA (p-hydroxybenzoic acid), chorismic acid and isochorismic acid of CoQ10 and VK2 in a two-step mixed fermentation process of vitamin C. By means of enhancement of the respiratory function of the ketogulonigenium sp., on one hand, the basal metabolic rate of the ketogulonigenium sp. can be increased, and the thallus quantity can be remarkably increased; on the other hand, the2-KGA producing speed can be increased due to the fact that a 2-KGA producing process of the ketogulonigenium sp. is coupled with the electron transport level of a respiratory chain electron transport process, and the fermentation period can be shortened by 15.38% to the maximum for a 5L fermentation tank. The method is advanced in production technology, low in raw material cost and suitable for large-scale industrial production, and is an improvement and upgrading of a biological fermentation process which is used for nearly 40 years for the vitamin C preparation; and further, emission of harmful substances in the production process is avoided, the surrounding environment is not contaminated, and the method is pollution-free and meets environmental requirements.

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 by selecting and optimizing microbial growth factors, and improving vitamin C fermentation craft. 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 acid (2-KGA), an important interm...

Claims

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Application Information

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IPC IPC(8): C12P39/00C12P7/60C12R1/01
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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