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2-keto-L-gulonic acid high tolerance type gluconobacteroxydans and application thereof in vitamin C fermentation production

A technology of gluconobacterium oxidizing and gulon acid, which is applied in the direction of microorganism-based methods, fermentation, bacteria, etc., can solve the problems of low fermentation efficiency and poor tolerance, and achieve the goal of improving production efficiency, reducing production costs and increasing production Effect

Active Publication Date: 2012-10-31
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 problem in the prior art that Gluconobacter oxydans has poor tolerance to 2-keto-L-gulonic acid, which leads to low fermentation efficiency in the second step of vitamin C production, and provides a 2-keto -L-gulonic acid high tolerance type Gluconobacter oxydans TL-1045, which has strong tolerance to 2-keto-L-gulonic acid, and has a high conversion efficiency of L-sorbose in the two-step fermentation, and the obtained Significantly increased yield of 2-keto-L-gulonic acid

Method used

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  • 2-keto-L-gulonic acid high tolerance type gluconobacteroxydans and application thereof in vitamin C fermentation production
  • 2-keto-L-gulonic acid high tolerance type gluconobacteroxydans and application thereof in vitamin C fermentation production
  • 2-keto-L-gulonic acid high tolerance type gluconobacteroxydans and application thereof in vitamin C fermentation production

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Effect test

Embodiment 1

[0044] Mutation Screening of Gluconobacter oxydans TL-1045

[0045] Pick out a single colony of the originating strain Gluconobacter oxydans TL-347, insert it into a sterile Erlenmeyer flask equipped with glass beads, add 30 ml of sterile saline, shake at 150 rpm for 20 min, break up the colony, and prepare bacterial suspension. Take 5 ml of bacterial suspension and add it to a petri dish, and irradiate it at 20-40 cm under a 40-watt ultraviolet lamp for 1 min, 5 min, and 10 min, and then dilute the three treated bacterial suspensions to 10 -3 -10 -5 , coated with selection medium (2-keto-L-gulonic acid 110 g / L, sorbose 15 g / L, tryptone 3 g / L, urea 0.1 g / L, corn steep liquor 4 g / L, beef cream 1.2 g / L, yeast extract 1.2 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.2 g / L, agar powder 17 g / L, pH 6.7), cultured in the dark for 72 h, picked The colonies on the plate were taken and paired with Bacillus megaterium respectively, and then the mixed bacteria were in...

Embodiment 2

[0048] The production of 2-keto-L-gulonic acid by mixed fermentation of Gluconobacter oxydans TL-1045 and Bacillus megaterium comprises the following steps:

[0049] (1) Activation of strains: Wash all the large and small bacteria stored in the eggplant bottle with the activation medium, inoculate them into the activation medium with a liquid volume of 20%, and incubate at 29°C for 24 h; the composition of the activation medium is For: sorbose 5 g / L, glucose 3 g / L, yeast extract 5 g / L, corn steep liquor 5 g / L, urea 0.1 g / L, pH 6.7;

[0050] (2) Separation and purification of strains: Under sterile conditions, use sterile physiological saline to gradiently dilute the strains activated in (1), take 10 -5 Spread 0.1 ml of the dilution on the separation plate, and culture the plate at 29°C for 6 days; the medium composition of the plate is: sorbose 15 g / L, tryptone 3 g / L, urea 0.1 g / L, corn steep liquor 4 g / L, beef extract 1.2 g / L, yeast extract 1.2 g / L, potassium dihydrogen phos...

Embodiment 3

[0056] The production of 2-keto-L-gulonic acid by mixed fermentation of Gluconobacter oxydans TL-1045 and Bacillus megaterium comprises the following steps:

[0057] (1) Activation of strains: Wash all the large and small bacteria stored in the eggplant bottle with the activation medium, inoculate into the activation medium with a liquid volume of 20%, and incubate at 31°C for 22 hours; the composition of the activation medium is For: sorbose 15 g / L, glucose 2 g / L, yeast extract 8 g / L, corn steep liquor 3 g / L, urea 0.05 g / L, pH 7.0;

[0058] (2) Separation and purification of strains: Under sterile conditions, use sterile physiological saline to gradiently dilute the strains activated in (1), take 10 -6 Spread 0.1 ml of the diluent on the separation plate, and culture the plate at 31°C for 4 days; the medium composition of the plate is: sorbose 10 g / L, tryptone 1 g / L, urea 0.2 g / L, corn steep liquor 6 g / L, beef extract 0.6 g / L, yeast extract 0.5 g / L, potassium dihydrogen phosph...

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Abstract

The invention discloses 2-keto-L-gulonic acid high tolerance type gluconobacteroxydans TL-1045 and also provides application of the bacterial strain in vitamin C fermentation production. The gluconobacteroxydans TL-1045 can tolerate the 2-keto-L-gulonic acid which has high concentration, the feedback inhibition of a product for a thallus in a second step during vitamin C fermentation production is eliminated, the output of 2-keto-L-gulonic acid can reach above 125g / L, the production efficiency of the vitamin C is improved remarkably and the production cost of the vitamin C is reduced effectively.

Description

technical field [0001] The present invention relates to a 2-keto-L-gulonic acid highly tolerant strain - Gluconobacter oxydans TL-1045 and its application in the fermentative production of vitamin C, specifically its use in the fermentative production of vitamin C The application in body 2-keto-L-gulonic acid and vitamin C belongs to the field of microbial fermentation 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...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P39/00C12P7/60C12P17/04C12R1/01C12R1/11
Inventor 张全景付吉明王乔隆刘敏郑秀宁庄祎
Owner SHANDONG TIANLI PHARMA
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