Method for detecting nutrition environment change in subculture process of vitamin C production strain

A technology for the production of bacterial strains and environmental changes, applied in the field of industrial microorganisms, can solve problems such as unclear mechanism of action, achieve the effect of increasing production and optimizing the fermentation process

Inactive Publication Date: 2012-08-15
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through subculture of mixed bacteria, the ability of mixed bacteria to ferment and produce 2-keto-L-gulonic acid is improved, but the mechanism of action is still unclear

Method used

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  • Method for detecting nutrition environment change in subculture process of vitamin C production strain
  • Method for detecting nutrition environment change in subculture process of vitamin C production strain
  • Method for detecting nutrition environment change in subculture process of vitamin C production strain

Examples

Experimental program
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Effect test

Embodiment 1

[0040] A method for detecting changes in the nutritional environment during the subculture of vitamin C production strains, comprising the steps of:

[0041] (1) Subculture of mixed bacteria:

[0042] ① Solid culture:

[0043] Inoculate 500 μL of Gluconobacter oxydans stored in 15% aqueous glycerol and 500 μL of Bacillus megaterium stored in 15% aqueous glycerol in liquid nitrogen, respectively. Incubate on solid medium at 28°C for 24 hours;

[0044] ②Seed cultivation:

[0045] Transfer the Bacillus megaterium and Gluconobacter oxidans cultivated in step (1) into the seed culture medium respectively, and cultivate them on a shaker at 28°C and 200r / min for 24 hours to obtain the Bacillus megaterium seed liquid and Gluconobacter oxidans seed liquid ;

[0046] Inoculate the new seed medium with Bacillus megaterium and Gluconobacter oxydans so that the density of Bacillus megaterium is 2×10 7 CFU / mL, so that the density of Gluconobacter oxidans is 2×10 8CFU / mL, at 28°C, 200r...

Embodiment 2

[0077] A method for detecting changes in the nutritional environment during the subculture of vitamin C production strains, comprising the steps of:

[0078] (1) Subculture of mixed bacteria:

[0079] ① Solid culture:

[0080] Take 10 μL of Gluconobacter oxydans (Gluconobacter oxydans) preserved in 20% aqueous glycerol solution and 10 μL of Bacillus megaterium (Bacillus megaterium) preserved in 20% aqueous glycerol solution and inoculate them respectively. Incubate on solid medium at 30°C for 36 hours;

[0081] ②Seed cultivation:

[0082] Transfer the Bacillus megaterium and Gluconobacter oxidans cultured in step (1) into the seed culture medium respectively, and cultivate them on a shaker at 30°C and 250r / min for 36 hours to obtain the Bacillus megaterium seed liquid and Gluconobacter oxidans seeds respectively liquid;

[0083] Inoculate a new seed medium with Bacillus megaterium and Gluconobacter oxydans so that the density of Bacillus megaterium is 2 x 10 8 CFU / mL, so ...

Embodiment 3

[0101] A method for detecting changes in the nutritional environment during the subculture of vitamin C production strains, comprising the steps of:

[0102] (1) Subculture of mixed bacteria:

[0103] ① Solid culture:

[0104] Take 200 μL of Gluconobacter oxydans (Gluconobacter oxydans) preserved in 30% aqueous glycerol solution and 200 μL of Bacillus megaterium (Bacillus megaterium) preserved in 30% aqueous glycerol solution stored in liquid nitrogen and inoculate them respectively Incubate on solid medium at 35°C for 48 hours;

[0105] ②Seed cultivation:

[0106] Transfer the Bacillus megaterium and Gluconobacter oxidans cultivated in step (1) ① into the seed culture medium respectively, and culture at 35°C and 280r / min shaking table for 48 hours to obtain the Bacillus megaterium seed liquid and Gluconobacter oxidans seed liquid ;

[0107] Inoculate a new seed medium with Bacillus megaterium and Gluconobacter oxydans so that the density of Bacillus megaterium is 2 x 10 ...

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Abstract

The invention discloses a method for detecting nutrition environment change in a subculture process of a vitamin C production strain. The method comprises the following steps of: subculture of mixed strains, detection on matters in a nutrition environment, analysis on main ingredients and analysis on the process. The method can be used for finding important matters influencing the subculture nutrient environment from revelation of the influence of the mixed strain subculture on bacillus megaterium, gluconobacter oxydans and mixed strain systems, and the change rules of the contents of the matters provide fundamental basis for understanding the mechanism of action of the subculture to promote the growth of gluconobacter oxydans and the production of 2-keto-L-gulonic acid hydrate, thus providing fundamental basis for further optimizing the fermentation process and improving the yield of vitamin C.

Description

technical field [0001] The invention belongs to the field of industrial microbes, and relates to a method for detecting nutritional environment changes in the process of subculture of vitamin C production strains. Background technique [0002] At present, the method of producing vitamin C in my country is a "two-step fermentation method". The first step of fermentation uses Acetobacter black to convert sorbitol into L-sorbose, and the second step of fermentation is a mixed fermentation of Bacillus megaterium and Gluconobacter oxidans. Converts sorbose to vitamin C precursor 2-keto-L-gulonic acid. Wherein, in the second step of fermentation, the former is an associated bacterium, and the latter is an acid-producing bacterium. In the process of mixed fermentation, the two bacteria promote the growth and acid production of acid-producing bacteria through interaction. Through subculture of mixed bacteria, the ability of mixed bacteria to ferment and produce 2-keto-L-gulonic aci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N30/02C12R1/11C12R1/01
Inventor 元英进高赟邹旸胡梦龙任恒千
Owner TIANJIN UNIV
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