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Method for preparing 2-keto-L-gulonic acid

A gulonic acid and ketone-based technology is applied in the field of preparation of vitamin C precursor - 2-keto-L-gulonic acid (2-KLG), which can solve the problems of low production efficiency, high efficiency, and short time. Achieve the effects of improving production efficiency, reducing production costs, and increasing product concentration

Active Publication Date: 2013-09-11
SHANDONG TIANLI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at the low production efficiency of the second step of fermentation in the existing vitamin C production process, and provides a method for preparing 2-keto-L-gulonic acid, in which sorbose is converted into 2-keto- L-gulonic acid has short time and high efficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] A preparation method of 2-keto-L-gulonic acid (2-KLG), comprising the following steps:

[0052] 1. The preparation and fermentation process of the large bacteria seed liquid are as follows:

[0053] (1) Activation of strains: Wash and inoculate the large bacteria culture medium stored in the eggplant bottle into a shaker flask filled with activation medium, fill with 20% liquid, culture at 31°C and 100 rpm for 14 hours, and activate The composition of the medium is: 3 g / L sorbose, 1 g / L glucose, 3 g / L tryptone, 3 g / L yeast extract, 4 g / L corn steep liquor, 2 g / L magnesium sulfate, pH 7.5;

[0054] (2) Purification of strains: gradiently dilute the activated strain solution obtained in step (1) with sterile physiological saline, take 10 -6 Spread 0.1 ml of the dilution on the plate, and then incubate the plate at 31°C for 36 hours. The plate medium consists of: sorbose 5g / L, tryptone 3g / L, urea 0.05 g / L, corn steep liquor 2 g / L , beef extract 2 g / L, yeast extrac...

Embodiment 2

[0069] A method for producing vitamin C precursor——2-keto-L-gulonic acid (2-KLG), the bacterial species used is the same as that in Example 1, comprising the following steps:

[0070] The preparation and fermentation process of the large bacteria seed liquid are as follows:

[0071] (1) Activation of strains: Wash and inoculate the large bacteria culture medium stored in the eggplant bottle into a shaker flask filled with activation culture medium. The composition of the medium is: sorbose 2 g / L, glucose 4 g / L, tryptone 4 g / L, yeast extract 2 g / L, corn steep liquor 6 g / L, magnesium sulfate 1 g / L, pH 7.0;

[0072] (2) Purification of strains: gradiently dilute the activated strain solution obtained in step (1) with sterile physiological saline, take 10 -7 Spread 0.1 ml of the dilution on the plate, and then incubate the plate at 30°C for 30 hours. The plate medium composition is: 6 g / L sorbose, 1 g / L tryptone, 0.1 g / L urea, 3 g / L corn steep liquor , beef extract 1.5 g...

Embodiment 3

[0087] A method for producing vitamin C precursor-2-keto-L-gulonic acid (2-KLG), the strain used is the same as in Example 1, comprising the following steps:

[0088] The preparation and fermentation process of the large bacteria seed liquid are as follows:

[0089] (1) Activation of strains: Wash and inoculate the large bacteria medium stored in the eggplant bottle into a shaker flask filled with activation medium, with a liquid volume of 20%, culture at 33°C and 150 rpm for 13 hours, and activate The medium composition is: sorbose 5 g / L, glucose 3 g / L, tryptone 2 g / L, yeast extract 2 g / L, corn steep liquor 2 g / L, magnesium sulfate 3 g / L, pH7.3;

[0090] (2) Purification of strains: gradiently dilute the activated strain solution obtained in step (1) with sterile physiological saline, take 10 -5 Spread 0.1 ml of the dilution on the plate, and then incubate the plate at 33°C for 26 hours. The plate medium composition is: 6 g / L sorbose, 1 g / L tryptone, 0.1 g / L urea, 3 ...

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Abstract

The invention discloses a method for preparing 2-keto-L-gulonic acid. The method is characterized by comprising the steps of: independently cultivating large bacteria, successively carrying out microfiltration and ultrafiltration treatment on the obtained large bacteria fermentation liquor to obtain an active substance concentrated solution; and feeding the obtained active substance concentrated solution of the large bacteria to a small bacteria culture medium to promote small bacteria to convert sorbose into 2-keto-L-gulonic acid. By adopting the method, the large bacteria are independently cultivated; preferably, the large bacteria are synchronously cultivated, so that the optimum growth condition of the large bacteria can be met; the production efficiency is improved; an active enzyme substance of the large bacteria is collected to transfer into small bacteria fermentation liquor by adopting advanced microfiltration and ultrafiltration technologies; bottom sugar and pH of the small bacteria fermentation liquor can be further stabilized by feeding the sorbose solution and a pH maintaining agent; the quantity of the small bacteria is increased; the growth time of thallus is shortened; the reaction rate of converting the sorbose oxidized by the small bacteria into 2-KLG is quickened; the fermentation cycle is shortened; the concentration of a product is improved; the production cost is reduced; and the market competitiveness of an enterprise is improved.

Description

technical field [0001] The present invention relates to a preparation method of a vitamin C precursor - 2-keto-L-gulonic acid (2-KLG), in particular to separately cultivating large and small bacteria under optimal conditions to produce 2-keto- The method for L-gulonic acid belongs to the technical field of vitamin C production. Background technique [0002] Vitamin C, also known as L-ascorbic acid, can participate in various redox reactions and hydroxylation reactions in the body. Blood disease, enhance the body's resistance. Green plants can synthesize vitamin C by themselves. However, humans and many animals cannot synthesize vitamin C by themselves due to the lack of gurolactone oxidase in their livers, and must ingest it from the outside. At present, vitamin C has been widely used in the fields of medicine, food, health care products and cosmetics. And with the development of its use and the improvement of people's material and cultural living standards, the demand fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/60C12R1/01C12R1/11C12R1/085C12R1/07C12R1/125
Inventor 庄祎王乔隆刘敏李慧君郑秀宁
Owner SHANDONG TIANLI PHARMA
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