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Method for fermentation production of 2-keto-L-ulonic acid

The technology of ketocoulonic acid bacillus and couronic acid is applied in the field of fermentation, which can solve the problems of high production cost, short fermentation period, long fermentation period and the like, and achieves the effects of not easy to contaminate bacteria, easy cultivation and simplified production process.

Inactive Publication Date: 2015-03-11
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of this, the object of the present invention is to provide a short fermentation cycle for the defects of long fermentation cycle, large energy consumption and high production cost for the production of vitamin C precursor 2-keto-L-gulonic acid by two-step fermentation in the prior art. Method for the fermentative production of 2-keto-L-gulonic acid

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  • Method for fermentation production of 2-keto-L-ulonic acid
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  • Method for fermentation production of 2-keto-L-ulonic acid

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Embodiment 1

[0053] Embodiment 1, the preparation of the culture medium of fermentative production 2-keto-L-gulonic acid

[0054] 1. Go bacteria seed medium:

[0055] D-sorbitol 2.0%, corn steep liquor 0.3%, beef extract 0.3%, yeast extract powder 0.3%, urea 0.1%, peptone 1.0%, KH 2 PO 4 0.1%, MgSO 4 ·7H 2 O 0.02%, CaCO 3 0.1%, the balance is water. Adjust the pH to 6.8, and sterilize at 121°C for 20 minutes.

[0056] 2. Kv bacteria seed medium:

[0057] L-sorbose 2.0%, corn steep liquor 0.3%, beef extract 0.3%, yeast extract powder 0.3%, urea 0.1%, peptone 1.0%, KH 2 PO 4 0.1%, MgSO 4 ·7H 2 O 0.02%, CaCO 3 0.1%, the balance is water. Adjust the pH to 6.8, and sterilize at 121°C (L-sorbose is sterilized alone, and added to the seed medium before inoculation) for 20 minutes.

[0058] 3. Fermentation medium:

[0059] D-Sorbitol 8.0%, Corn Steep Steep 1.0%, Urea 1.2%, KH 2 PO 4 0.1%, MgSO 4 ·7H 2 O0.02%, CaCO 3 0.1%, the balance is water. Adjust the pH to 6.8-7.0, and ster...

Embodiment 2

[0060] Embodiment 2, production of 2-keto-L-gulonic acid by fermentation

[0061] 1. Seed cultivation

[0062] Primary seed culture: Take 300 μl of glycerol bacteria and add it to 50 ml of seed medium, and culture at 30°C and 250 rpm on a shaker for 24 hours.

[0063] Secondary seed culture: According to the inoculation ratio of 10%, take 5ml of primary seeds and inoculate them into 50ml of seed medium, and culture them on a shaker at 250rpm at 30°C for 24h.

[0064] 2. Fermentation tank culture

[0065] According to the conditions of 10% inoculum amount and bacterium ratio Go:Kv=5:1, a total of 300ml of secondary seed liquid was inserted into a 5L fermenter (filling factor 60%). The initial input material is 4.0% D-sorbitol, after 3 hours, start to feed the remaining 4%, and ferment for about 48 hours under the conditions of cultivation temperature 30°C, stirring speed 450rpm, ventilation rate 1.5vvm.

Embodiment 3

[0066] Embodiment 3, production of 2-keto-L-gulonic acid by fermentation

[0067] 1. Seed cultivation

[0068] With embodiment 2.

[0069] 2. Fermentation tank culture

[0070] According to the conditions of 10% inoculum amount and bacterium ratio Go:Kv=5:1, a total of 300ml of secondary seed liquid was inserted into a 5L fermenter (filling factor 60%). The initial input material is 4.0% D-sorbitol, after 3 hours, start to feed the remaining 4%, and ferment for 28 hours under the conditions of cultivation temperature 30°C, stirring speed 500rpm, ventilation rate 1.5vvm.

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Abstract

The invention relates to the technical field of fermentation, and discloses a method for fermentation production of vitamin C precursor 2-keto-L-ulonic acid. The method is that the mixing of Go fungus and Kv fungus forms one-step coversion from D-sorbitol to 2-keto-L-ulonic acid, simplifies the production technology, shortens the fermentation cycle, and reduces the production cost. Meanwhile, the mixed culture of the Go fungus and the Kv fungus is simple, is not easy to cause microbiological contamination, is stable, can utilize an original fermentation tank to perform fermentation, and is easy for realizing industrialization. Furthermore, the method for fermentation production of precursor 2-keto-L-ulonic acid adopts gluconobacter oxydans, with shunt metabolism-related genes of sorbose / sorbitol being knocked out, to be matched with the Kv to subjected to mixed fermentation, prevents the Go fungus and the Kv fungus from competitively fighting for substrate sorbose to a certain degree, and improves the conversion efficiency.

Description

technical field [0001] The invention relates to the technical field of fermentation, in particular to a method for fermenting and producing vitamin C, in particular to a method for fermenting and producing vitamin C precursor 2-keto-L-gulonic acid and related fermentation strains. Background technique [0002] Vitamin C (Vitamin C, VC), also known as L-ascorbic acid (L-ascorbic acid), is an essential nutrient for the human body. Vitamin C is for the formation of antibodies and collagen, tissue repair (including certain redox effects), metabolism of phenylalanine, tyrosine, and folic acid, utilization of iron and carbohydrates, synthesis of fat and protein, maintenance of immune function, hydroxyl Vitamin C is necessary for oxidizing and serotonin, maintaining the integrity of blood vessels, and promoting the absorption of non-heme iron. As an antioxidant, vitamin C plays an important role in regulating redox metabolic reactions in the human body. At the same time, vitamin C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P7/60C12N1/20C12R1/01
Inventor 元英进王恩旭丁明珠
Owner TIANJIN UNIV
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