Biotin supplemented fermentation method for producing glutamic acid

A fermentation method and biotin technology, applied in microorganism-based methods, biochemical equipment and methods, fermentation and other directions, can solve the problem of weakened pyruvate carboxylase activity, reduced glutamic acid fermentation rate, lack of biotin in bacterial cells, etc. problem, to achieve the effect of good cell permeability, easy control of addition time, and convenient control

Active Publication Date: 2014-12-03
COFCO BIO CHEM ENERGY LONGJIANG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the case of limited biotin, the biotin in the fermentation medium is exhausted in the middle and later stages of fermentation, and the bacteria lack biotin, which leads to the key reaction in the synthesis of glutamic acid (that is, the carboxylation of pyruvate to oxaloacetate). The activity of pyruvate carboxylase is weakened, which leads to the decrease of glutamic acid fermentation rate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] There are three kinds of medium used in this embodiment:

[0040] ⑴Seed medium: glucose 26g / L, corn steep liquor 10g / L, molasses 5g / L, MgSO 4 ·7H 2 O 0.9g / L, KCl 1.2g / L, MnSO 4 0.00025g / L, V B1 0.0002g / L, Na 2 HPO 4 1.5g / L, pure biotin 0μg / L, pH 7.0, sterilized at 120°C for 20 minutes;

[0041] ⑵Fermentation medium: glucose 130g / L, corn steep liquor 2.5g / L, molasses 1.5 g / L, MgSO 4 ·7H 2 O 0.5g / L, KCl 2.5g / L, MnSO 4 0.006g / L, V B1 0.0005g / L, Na 2 HPO 4 3.5g / L, pure biotin 14μg / L, foam enemy 0.3g / L, pH 7.0, sterilized at 115°C for 20 minutes;

[0042] (3) Supplementary sugar solution: Supplementary sugar solution is glucose solution with a mass concentration of glucose of 600 g / L, sterilized at 115°C for 20 minutes.

[0043] A fermentation method for producing glutamic acid supplemented with biotin, the steps are as follows:

[0044] (1) Insert the biotin-deficient Corynebacterium glutamicum S9114 on the one-ring slant medium into a sterilized 1L Erle...

Embodiment 2

[0050] There are three kinds of medium used in this embodiment:

[0051] (1) Seed medium: glucose 35g / L, corn steep liquor 11g / L, molasses 5g / L, MgSO 4 ·7H 2 O 0.6g / L, KCl 1.0g / L, MnSO 4 0.00025g / L, V B1 0.0001g / L, Na 2 HPO 4 2.5g / L, pure biotin 0μg / L, pH 7.0, sterilized at 120°C for 20 minutes.

[0052] (2) Fermentation medium: glucose 130g / L, corn steep liquor 2.5g / L, molasses 1.5 g / L, MgSO 4 ·7H 2 O 0.5g / L, KCl 2.5g / L, MnSO 4 0.003g / L, V B1 0.0005g / L, Na 2 HPO 4 3.5g / L, pure biotin 24μg / L, foam enemy 0.3g / L, pH 7.0, sterilized at 115°C for 20 minutes.

[0053] (3) Supplementary sugar: Supplementary sugar solution is glucose solution with a mass concentration of glucose of 600 g / L, sterilized at 115°C for 20 minutes.

[0054] A fermentation method for producing glutamic acid supplemented with biotin, the steps are as follows:

[0055] (1) Insert biotin-deficient Corynebacterium glutamicum S9114 on a ring of slant medium into a sterilized 1L Erlenmeyer flask...

Embodiment 3

[0062] There are three kinds of medium used in this embodiment:

[0063] (1) Seed medium: glucose 20g / L, corn steep liquor 13g / L, molasses 5g / L, MgSO 4 ·7H 2 O 0.7g / L, KCl 1.0g / L, MnSO 4 0.00025g / L, V B1 0.0001g / L, Na 2 HPO 4 1.5g / L, pure biotin 0μg / L, pH 7.0, sterilized at 120°C for 20 minutes.

[0064] (2) Fermentation medium: glucose 130g / L, corn steep liquor 2.5g / L, molasses 1.5 g / L, MgSO 4 ·7H 2 O 0.5g / L, KCl 2.5g / L, MnSO 4 0.003g / L, V B1 0.0005g / L, Na 2 HPO 4 3.5g / L, pure biotin 24μg / L, foam enemy 0.3g / L, pH 7.0, sterilized at 115°C for 20 minutes.

[0065] (3) Supplementary sugar: Supplementary sugar solution is glucose solution with a mass concentration of glucose of 600 g / L, sterilized at 115°C for 20 minutes.

[0066] A fermentation method for producing glutamic acid supplemented with biotin, the steps are as follows:

[0067] (1) Insert the biotin-deficient Corynebacterium glutamicum S9114 on the one-ring slant medium into a sterilized 1L Erlenmeye...

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Abstract

The invention relates to a biotin supplemented fermentation method for producing glutamic acid. The method comprises the following steps: treating biotin deficient strains as production strains, controlling the initial content of biotins in a fermentation culture medium, and supplementing penicillin and biotins in the fermentation process. The fermentation method which allows penicillin and biotins to be added in the middle and later fermentation process can overcome a biotin poorness problem in the later fermentation period of a suboptimal-dose technology, guarantees the sufficient biotins in the culture medium for the growth and the metabolism of thalli, and substantially improves the glutamic acid synthesis speed of the thalli; and compared with present glutamic acid suboptimal-dose technologies, the fermentation method provided by the invention allows the acid production rate of unit thalli to be improved by above 10%, the fermentation period to be substantially shortened and the production cost to be reduced.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation engineering and relates to the production of glutamic acid, in particular to a fermentation method for producing glutamic acid by adding biotin. Background technique [0002] At present, most domestic glutamic acid production enterprises adopt biotin-deficient strains and adopt biotin "sub-appropriate" or "super-sub-appropriate" fermentation process. However, both of these two fermentation processes require the content of biotin in the fermentation medium to be controlled and cannot exceed the limit level, resulting in a serious lack of biotin in the fermentation medium in the middle and late stages of fermentation, and the cell viability and acid production rate are limited. It can be seen that there is a problem of biotin deficiency in the "sub-appropriate" or "super-sub-appropriate" glutamic acid fermentation process, which is not conducive to improving the rate of fermentation ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/18C12R1/15
Inventor 俞建良耿兴山王晓健贺玉明王晓丽白长胜
Owner COFCO BIO CHEM ENERGY LONGJIANG CO LTD
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