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An analysis method for a Corynebacterium glutamicum metabolome

A Corynebacterium glutamicum, analysis method technology, applied in the field of bioengineering, to achieve the effect of promoting metabolism research

Inactive Publication Date: 2018-11-09
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on metabolomics related to Corynebacterium glutamicum, so it is extremely important to study the changes of related metabolites during the fermentation process of Corynebacterium glutamicum

Method used

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  • An analysis method for a Corynebacterium glutamicum metabolome
  • An analysis method for a Corynebacterium glutamicum metabolome
  • An analysis method for a Corynebacterium glutamicum metabolome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A method for analyzing the metabolome of Corynebacterium glutamicum, comprising the steps of:

[0036] (1) Corynebacterium glutamicum (Corynebacterium glutamicum) XV intracellular metabolite treatment:

[0037] ① Cell collection and quenching

[0038] A total of 5 sampling points were taken at the early stage, middle stage, and late stage of Corynebacterium glutamicum XV fermentation, and 50ml of fermentation broth was taken for each, and centrifuged at 4°C for 3min to remove most of the solid impurities in the medium. Then, centrifuge at 8000rmp, 4°C for 10min. The supernatant was discarded, and the cell pellet was washed twice with PBS buffer, and then once with deionized ultrapure water. After centrifugation, the cell pellet was placed in liquid nitrogen to quench the reaction, and stored in liquid nitrogen for later use.

[0039] Described PBS damping fluid (1mol / L) contains NaCl 8g, KCl 0.2g, NaCl 2 HPO 4 1.42g, KH 2 PO 4 0.27g

[0040] ②Metabolite extract...

Embodiment 2

[0063] A method for analyzing the metabolome of Corynebacterium glutamicum, comprising the steps of:

[0064] (1) Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC 13032 intracellular metabolite treatment:

[0065] ① Cell collection and quenching

[0066] A total of 5 sampling points were taken at the early stage, middle stage and late stage of the fermentation of Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC 13032, and 60ml of fermentation broth was taken for each, and centrifuged at 4°C for 3min to remove most of the solid impurities in the medium. Then, centrifuge at 7000rmp and 4°C for 10min. The supernatant was discarded, and the cell pellet was washed twice with PBS buffer, and then once with deionized ultrapure water. After centrifugation, the cell pellet was placed in liquid nitrogen to quench the reaction, and stored in liquid nitrogen for later use.

[0067] Described PBS damping fluid (1mol / L) contains NaCl 8g, KCl 0.2g, NaCl 2 HPO 4 1...

Embodiment 3

[0077] A method for analyzing the metabolome of Corynebacterium glutamicum, comprising the steps of:

[0078] (1) Corynebacterium glutamicum (Corynebacterium glutamicum) CP intracellular metabolite treatment:

[0079] ① Cell collection and quenching

[0080] A total of 5 sampling points were taken at the early stage, middle stage, and late stage of CP fermentation of Corynebacterium glutamicum (Corynebacterium glutamicum), and 50ml of fermentation broth was taken for each, and centrifuged at 4°C for 3min to remove most of the solid impurities in the medium. Then, centrifuge at 8000rmp, 4°C for 10min. The supernatant was discarded, and the cell pellet was washed twice with PBS buffer, and then once with deionized ultrapure water. After centrifugation, the cell pellet was placed in liquid nitrogen to quench the reaction, and stored in liquid nitrogen for later use.

[0081] Described PBS damping fluid (1mol / L) contains NaCl 8g, KCl 0.2g, NaCl 2 HPO 4 1.42g, KH 2 PO 4 0.2...

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Abstract

An analysis method for a Corynebacterium glutamicum metabolome is provided. The method includes Corynebacterium glutamicum cell collection and quenching; intracellular metabolite extraction; metabolite derivatization; and sample analysis through GC-Ms and multivariate statistics to obtain a metabolome result finally. Analysis measures for the Corynebacterium glutamicum metabolome relate to metabolic reaction terminating, metabolite extraction, metabolite identification and quantitative and statistical analysis, thus obtaining qualitative and quantitative results of metabolites in different stages in a Corynebacterium glutamicum fermentation process, and providing a way for analysis of the Corynebacterium glutamicum metabolome. The method is of great significance for promoting metabonomic and metabolic engineering researches of Corynebacterium glutamicum.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for analyzing the metabolome of Corynebacterium glutamicum. Background technique [0002] Corynebacterium glutamicum is an aerobic, non-spore-forming, Gram-positive bacterium with short, rod-shaped cells, blunt ends, and a figure-eight arrangement. It does not move, and the colonies are moist and round. As a food-grade microorganism, Corynebacterium glutamicum has the advantages of easy cultivation and no spore production. Therefore, it is an important class of industrial production strains and can be used to produce various amino acids and antibiotics. At present, the traditional mutagenesis method is still the main way to obtain industrial production strains of Corynebacterium glutamicum, but due to its unknown genetic background, it is difficult to increase amino acid production through further mutagenesis. [0003] It can be seen that with the continuous maturity of va...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 马倩莫晓琳陈宁谢希贤侯正杰张权威徐庆阳李燕军张成林范晓光
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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