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Single-strain preparation and fermentation technology based on fermentation of 2-keto-L-gulonic acid

A technology for preparing ketogenic cologne bacteria and strains, which is applied in the directions of fermentation, microorganism-based methods, biochemical equipment and methods, etc. Improvement, screening and inconvenience of preservation, etc.

Inactive Publication Date: 2013-05-22
DSM JIANGSHAN PHARMACEUTICAL (JIANGSU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, due to the mixed culture, it is extremely inconvenient for the genetic improvement, screening and preservation of small bacteria, and it is also not conducive to the sterility management of the strain culture and fermentation process

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0008] First inoculate the small bacteria individually into the eggplant bottle slant medium with a capacity of 70ml, and add 5ml of large bacteria lysate at the same time, cultivate for 72 hours, and use the freeze-vacuum drying method to make the cultured pure small bacterial slant and store it in a frozen tube. After 2 years of cryopreservation in cryogenic tubes, the survival rate was over 85%.

[0009] After activating and culturing the frozen tubes of preserved small bacteria through liquid seed medium for 40 hours, dilute the activated seed solution to 10 -7 , smeared and inoculated on the separation plate, cultured for 72 hours, and a single small bacterial colony was generated, and a number of excellent small bacteria were selected, inoculated into the eggplant bottle slant medium with a capacity of 70ml, cultivated for 72 hours, and the cultivated pure small bacteria The slant was inoculated into the liquid seed culture medium, and cultured for 24 hours to obtain a p...

Embodiment 2

[0012] First inoculate the small bacteria individually into the eggplant bottle slant medium with a capacity of 70ml, and add 5ml of large bacteria lysate at the same time, cultivate for 72 hours, and use the freeze-vacuum drying method to make the cultured pure small bacterial slant and store it in a frozen tube. After 2 years of cryopreservation in cryogenic tubes, the survival rate was over 85%.

[0013] After activating and culturing the frozen tubes of preserved small bacteria through liquid seed medium for 40 hours, dilute the activated seed solution to 10 -7 , smeared and inoculated on the separation plate, cultured for 72 hours, and a single small bacterial colony was generated, and a number of excellent small bacteria were selected, inoculated into the eggplant bottle slant medium with a capacity of 70ml, cultivated for 72 hours, and the cultivated pure small bacteria The slant was inoculated into the liquid seed culture medium, and cultured for 22 hours to obtain...

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PUM

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Abstract

The invention provides a single-strain preparation and fermentation technology based on the fermentation of 2-keto-L-gulonic acid. According to the technology, in the strain preparation and fermentation process, the bacillus megatherium and the ketogulonicigenium vulgaer are separately cultured, the bacillus megatherium is cultured in a liquid medium until reaching the end point so as to obtain the cell lysis solution for standby use; and the independently stored ketogulonicigenium vulgaer is sequentially activated and cultured, and isolated and screened, and is continuously inoculated to an inclined surface to enlarge in cultivation together with seed solution, and then is transferred into a fermentation medium to ferment. The bacillus megatherium is respectively added to the culture at each level, of the ketogulonicigenium vulgaer based on the 3 to 30% of the inoculum size, so that the normal growth and metabolism of the ketogulonicigenium vulgaer is promoted. By adopting the technology, the negative effect of a large strain to a small strain during mixing the large strain with the small strain to culture and ferment in the conventional technology can be avoided, the benefits are provided for genetic improvement, screening, culturing and storage of the small strain and management in the strain growth and metabolism process, and the fermentation period is reduced.

Description

technical field [0001] The invention relates to a preparation method of vitamin C, in particular to a 2-keto-L-gulonic acid fermentation single strain preparation and fermentation process. technical background [0002] At present, my country's vitamin C manufacturers all adopt two-step fermentation production technology. Its main feature is that the two-step fermentation process is completed by the mixed culture of two kinds of bacteria. Among them, common ketogenic cologne converts sorbose into 2-KLG. Ordinary ketogenic cologne, commonly known as small bacteria, is an acid-producing bacterium. The other type of bacteria is Bacillus megaterium, which promotes the growth and acid production of small bacteria. Bacillus megaterium, commonly known as large bacteria, is an associated bacterium. Both bacteria work together to convert sorbose to 2-KLG. In the known production technology of 2-KLG fermentation strains: from the activation of preserved strains to the expansion ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/60C12R1/01C12R1/11
Inventor 刘杰胡少斌陈红常芳
Owner DSM JIANGSHAN PHARMACEUTICAL (JIANGSU) CO LTD
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