Method for detecting change of proteins inside cells in industrial mixed fermentation process of vitamin C

A technology of mixed bacteria fermentation and fermentation process, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve the problems of increasing the difficulty of fermentation process, unclear internal regulation mechanism, and difficulty in optimizing industrial production process, etc., to achieve Increase yield, reduce environmental pollution, optimize the effect of fermentation process

Active Publication Date: 2011-09-07
TIANJIN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The industrial mixed bacterial fermentation of vitamin C is completed by the joint action of Bacillus megaterium and Gluconobacter oxydans. The internal regulation mechanism is still unclear, which makes it difficult to optimize the industrial production process and increases the need to control the fermentation process during production. difficulty

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  • Method for detecting change of proteins inside cells in industrial mixed fermentation process of vitamin C
  • Method for detecting change of proteins inside cells in industrial mixed fermentation process of vitamin C
  • Method for detecting change of proteins inside cells in industrial mixed fermentation process of vitamin C

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Embodiment 1

[0036] A method for detecting intracellular protein changes during vitamin C industrial mixed bacterial fermentation, comprising the following steps:

[0037] (1) Determination of proteins in cells of Bacillus megaterium CGMCC No 1.459 and Gluconobacter oxydans CGMCC No 1.110 used in the vitamin C fermentation process:

[0038] ① Cell collection and quenching:

[0039] Ferment the Bacillus megaterium and Gluconobacter oxydans, select 3-5 time points during the fermentation process, the time points are located in the early, middle and late stages of the fermentation process, and quickly take out the fermentation broth at the time points The sample was centrifuged, and the cells in the lower layer were collected, washed with phosphate buffered saline, and immediately quenched with liquid nitrogen to terminate the metabolic reaction; ground with liquid nitrogen for broken cells, and 5 parts of broken cells were obtained;

[0040] ② Extraction of intracellular protein:

[0041] ...

Embodiment 2

[0073] A method for detecting intracellular protein changes during vitamin C industrial mixed bacterial fermentation, comprising the following steps:

[0074] (1) Determination of proteins in cells of Bacillus megaterium CGMCC No 1.459 and Gluconobacter oxydans CGMCC No 1.110 used in the vitamin C fermentation process:

[0075] ① Cell collection and quenching:

[0076] Ferment the Bacillus megaterium and Gluconobacter oxydans, select 3-5 time points during the fermentation process, the time points are located in the early, middle and late stages of the fermentation process, and quickly take out the fermentation broth at the time points The sample was centrifuged, and the cells in the lower layer were collected, washed with phosphate buffered saline, quenched with liquid nitrogen immediately, and the metabolic reaction was terminated; ground with liquid nitrogen for breaking the cells, and 4 parts of broken cells were obtained;

[0077] ③ Determination of protein concentration...

Embodiment 3

[0095] A method for detecting intracellular protein changes during vitamin C industrial mixed bacterial fermentation, comprising the following steps:

[0096] (1) Determination of proteins in cells of Bacillus megaterium CGMCC No 1.459 and Gluconobacter oxydans CGMCC No 1.110 used in the vitamin C fermentation process:

[0097] ① Cell collection and quenching:

[0098] Ferment the Bacillus megaterium and Gluconobacter oxydans, select 3-5 time points during the fermentation process, the time points are located in the early, middle and late stages of the fermentation process, and quickly take out the fermentation broth at the time points The sample was centrifuged, and the cells in the lower layer were collected, washed with phosphate buffered saline, and immediately quenched with liquid nitrogen to terminate the metabolic reaction; ground with liquid nitrogen to break the cells, and 3 parts of broken cells were obtained;

[0099] ③ Determination of protein concentration:

[0...

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Abstract

The invention discloses a method for detecting the change of proteins inside cells in the industrial mixed fermentation process of vitamin C, comprising the following steps of: (1) measuring the proteins inside cells, such as huge Bacillus megaterium and Gluconobacter oxydans which are used for the industrial mixed fermentation process of the vitamin C; (2) carrying out cluster analysis; and (3) carrying out process analysis. The method can be used for integrally researching the change law of the proteins inside the cells in the industrial mixed fermentation process of the vitamin C and finding the important proteins in the industrial mixed fermentation process according to the change law of the proteins inside the cells; the change law of the content of the proteins provides basis for the inner mechanism of the industrial mixed fermentation process so as to provide theoretical basis for further optimizing the industrial mixed fermentation process, increasing the yield of the vitamin C and reducing the environmental pollution; in addition, the invention also provides a new thought and method for the research of other mixed fermentation processes.

Description

technical field [0001] The invention belongs to the field of industrial microorganisms, and relates to a method for detecting intracellular protein changes in vitamin C industrial mixed bacteria fermentation. Background technique [0002] Protein is an important component of cells, the regulator and executor of physiological functions, and the direct embodiment of life phenomena. The study of protein structure and function will directly clarify the mechanism of changes in cells under different growth conditions. On the one hand, it can verify its upstream genomics information, and on the other hand, it can provide guidance for its downstream metabolite research. The development of proteomics mainly relies on efficient protein separation and identification technology. Two-dimensional gel electrophoresis is currently the most classic protein separation method, which can separate hundreds or even thousands of proteins on one gel. At the same time, the development of mass spectr...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/27G01N27/26G01N27/62G01N1/28G01N1/38C12Q1/00C12R1/11C12R1/01
Inventor 元英进马倩米造吉谢萍
Owner TIANJIN UNIV
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