Method for analyzing change rule of metabolites during fermentation of cephalosporium acremonium

A technology of fermentation process and change law, applied in the direction of analysis of materials, material separation, measurement devices, etc., can solve problems such as loss of valuable information, single information, etc., to achieve the effect of complete analysis

Active Publication Date: 2012-11-21
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The information obtained by traditional data processing methods is relatively single, and still requires a lot of manual operations, which often results in the loss of a lot of valuable information

Method used

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  • Method for analyzing change rule of metabolites during fermentation of cephalosporium acremonium
  • Method for analyzing change rule of metabolites during fermentation of cephalosporium acremonium
  • Method for analyzing change rule of metabolites during fermentation of cephalosporium acremonium

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Embodiment 1

[0028] The method for analyzing the change rule of metabolites in the fermentation process of Cephalosporium acremonium comprises the following steps:

[0029] (1) Take 3 sampling points at the early stage, middle stage, and late stage of fermentation of the cephalosporin C production strain Cephalosporium acremonium CICC No 40515, take 100 ml of fermentation broth and centrifuge, and discard the supernatant.

[0030] (2) Add 100ml of phosphate buffer solution with a pH value of 7.2 to the precipitated part, mix well and centrifuge. The precipitated cells were quenched in liquid nitrogen to terminate the metabolic reaction of the cells, and ground to a fine powder with liquid nitrogen;

[0031] (3) Put 20 mg of finely powdered cells in a centrifuge tube, add 0.5 ml of methanol aqueous solution with a volume concentration of 50% stored at -40°C as the extract, and add 10 μl of 0.14 mg / ml deuterium-labeled succinic acid aqueous solution As an internal standard, mix well; place ...

Embodiment 2

[0041] The method for analyzing the change rule of metabolites in the fermentation process of Cephalosporium acremonium comprises the following steps:

[0042](1) Take 3 sampling points respectively at the early stage, middle stage and late stage of fermentation of cephalosporin C production strain Cephalosporium acremonium CICC No 40514, take 200ml of fermentation broth and centrifuge, and discard the supernatant.

[0043] (2) Add 200ml of phosphate buffer solution with a pH value of 7.2 to the precipitated part, mix well and centrifuge. The pelleted cells were quenched in liquid nitrogen to stop the metabolic reaction of the cells, and ground to a fine powder with liquid nitrogen.

[0044] (3) Place 50 mg of finely powdered cells in a centrifuge tube, add 1 ml of 50% aqueous methanol with a volume concentration of 45° C. as an extract, and add 15 μl of 0.14 mg / ml deuterium-labeled succinic acid aqueous solution as The internal standard was mixed evenly; placed in liquid nit...

Embodiment 3

[0050] The method for analyzing the change rule of metabolites in the fermentation process of Cephalosporium acremonium comprises the following steps:

[0051] (1) Take 3 sampling points at the early stage, middle stage, and late stage of fermentation of the cephalosporin C production strain Cephalosporium acremonium CICC No 2444, take 300ml of fermentation broth and centrifuge, and discard the supernatant.

[0052] (2) Add 300 ml of phosphate buffer solution with a pH value of 7.2 to the precipitated part, mix well and centrifuge. The pelleted cells were quenched in liquid nitrogen to stop the metabolic reaction of the cells, and ground to a fine powder with liquid nitrogen.

[0053] (3) Put 80 mg of finely powdered cells in a centrifuge tube, add 1.5 ml of 50% methanol aqueous solution stored at -50°C as the extract, and add 20 μl of 0.14 mg / ml deuterium-labeled succinic acid aqueous solution As an internal standard, mix well; place in liquid nitrogen and freeze-thaw 4 time...

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Abstract

The invention discloses a method for analyzing a change rule of metabolites during fermentation of cephalosporium acremonium. The method comprises the following steps of: sampling and centrifuging before fermentation, during fermentation and after fermentation of cephalosporium acremonium generated by cephalosporin C, mixing phosphate buffer solution into sediment uniformly and centrifuging; quenching by using liquid nitrogen and grinding; adding powdered cells into methanol aqueous solution and butanedioic acid aqueous solution which is marked by deuterium and is used as an internal standardsubstance, and mixing uniformly; freezing and melting, centrifugally collecting the supernate and drying; performing oximation reaction; performing silanization reaction; determining components and relative content of a sample by using a gas chromatography-flying time mass spectrometer; analyzing principal components of preprocessed data by using software so as to analyze metabolism difference ofthe samples; performing cluster analysis on the preprocessed data by using the software so as to analyze the metabolism difference of metabolites among the samples; determining the nature of the metabolites to classify the metabolites; and analyzing the change rule of all types of the metabolites during the fermentation so as to comprehensively disclose metabolism information of the cephalosporium acremonium according to a large amount of metabolism physiome data.

Description

technical field [0001] The invention relates to a method for analyzing the change rule of metabolites in the fermentation process of Cephalosporium acremonium. Background technique [0002] Cephalosporin antibiotics are a kind of β-lactam antibiotics, which achieve the purpose of sterilization by inhibiting the formation of bacterial cell walls. Because of its high antibacterial infection activity and low incidence of side effects, it is widely used in clinical practice. As a raw material for the synthesis of cephalosporins, the achievements in the research on the fermentation of cephalosporin C are far less than the development of a new generation of cephalosporins. Therefore, more research on the production process of cephalosporins is needed to obtain greater fermentation titers of cephalosporins. [0003] Metabolomics can study biological systems by examining the changes in their metabolites after the biological system is stimulated or disturbed. The key to metabolomi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
Inventor 元英进杨扬卢华陈尧
Owner TIANJIN UNIV
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