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

A technology of fermentation process and changing rules, applied in the direction of analyzing materials, material separation, measuring devices, etc., can solve the problems of valuable information loss, single information, etc., and achieve the effect of complete analysis

Active Publication Date: 2011-11-30
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

Examples

Experimental program
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Effect test

Embodiment 1

[0028] The analysis method of the changes of metabolites during the fermentation of Cephalosporium acremonium includes the following steps:

[0029] (1) In the cephalosporium C-producing strain Cephalosporium acremonium CICC No 40515, 3 sampling points were taken each in the early, middle and late stages of fermentation, 100ml of fermentation broth was centrifuged, and the supernatant was discarded.

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

[0031] (3) Place 20 mg of fine powdered cells in a centrifuge tube, add 0.5 ml of 50% methanol aqueous solution stored at -40°C as the extract, and add 10 μl of 0.14 mg / ml deuterium-labeled succinic acid aqueous solution It is an internal standard substance, mix well; place it in liquid nitrogen and freeze-thaw repeatedly twice, freezing ...

Embodiment 2

[0041] The analysis method of the changes of metabolites during the fermentation of Cephalosporium acremonium includes the following steps:

[0042] (1) In the cephalosporium C-producing strain Cephalosporium acremonium CICC No 40514, 3 sampling points were taken each in the early, middle and late stages of fermentation, 200ml of fermentation broth was centrifuged, and the supernatant was discarded.

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

[0044] (3) Place 50mg of fine powdered cells in a centrifuge tube, add 1ml-45℃ of 50% methanol aqueous solution as the extract, and add 15μl of 0.14mg / ml deuterium-labeled succinic acid aqueous solution. Mix the internal standard substance; place it in liquid nitrogen and freeze-thaw for 3 times, freeze each time for 1.5min; centrifuge to...

Embodiment 3

[0050] The analysis method of the changes of metabolites during the fermentation of Cephalosporium acremonium includes the following steps:

[0051] (1) In the cephalosporium C-producing strain Cephalosporium acremonium CICC No 2444, 3 sampling points were taken each at the early, middle and late stages of fermentation, 300ml of fermentation broth was centrifuged, and the supernatant was discarded.

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

[0053] (3) Put 80mg of fine powdered cells in a centrifuge tube, add 1.5ml of 50% methanol aqueous solution stored at -50°C as the extract, and add 20μl of 0.14mg / ml deuterium-labeled succinic acid aqueous solution It is an internal standard substance, mix well; place it in liquid nitrogen for 4 times of repeated freezing and thawing,...

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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 standard substance, 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 of the 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 law 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 anti-bacterial infection activity and low incidence of side effects, it is widely used in clinical practice. As a raw material for the synthesis of cephalosporin drugs, the achievements in the fermentation research of cephalosporin C are far less than the development of a new generation of cephalosporin drugs. Therefore, more research on the production process of cephalosporin is needed to obtain greater fermentation titer of cephalosporin. [0003] Metabolomics can study biological systems by investigating the changes in their metabolites after being stimulated or disturbed. The key to metabolomics lies in the process...

Claims

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Application Information

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