Method for obtaining metabolic flux of intracellular central carbon metabolic pathway in unsteady state of metabolic steady-state isotopes

A technology of metabolic pathways and isotopes, applied in the field of calculation of metabolic flux, can solve the problems of high cost, empty window period of flow addition of substrates, interference of microorganisms, etc., and achieve the effect of accurate calculation.

Pending Publication Date: 2020-08-25
EAST CHINA UNIV OF SCI & TECH
View PDF3 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the theoretical method for calculating metabolic flux is very mature, but the relevant experimental method is not perfect, and there are the following problems: the large-capacity bioreactor will waste unnecessary energy during the isotope labeling experiment. 13 C glucose, which leads to high experimental costs; in the experimental operation of switching substrates, complex pipelines are likely to cause a window period for feeding substrates, which affects the physiological and meta

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for obtaining metabolic flux of intracellular central carbon metabolic pathway in unsteady state of metabolic steady-state isotopes
  • Method for obtaining metabolic flux of intracellular central carbon metabolic pathway in unsteady state of metabolic steady-state isotopes
  • Method for obtaining metabolic flux of intracellular central carbon metabolic pathway in unsteady state of metabolic steady-state isotopes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] This embodiment provides a method for obtaining the metabolic flux of the intracellular central carbon metabolic pathway under the metabolic stable isotope unstable state, including a batch culture module, a chemostat culture module, a sampling module, and a processing and analysis sample module; wherein ,

[0046] When the oxygen uptake rate (OUR) and CO in the batch culture module 2 Transfer to the chemostat culture module when the release rate (CER) declines simultaneously;

[0047] The chemostat culture module specifically includes: feeding a nutrient solution without an isotope-labeled substrate, simultaneously turning on waste discharge, and performing a chemostat culture until the microbial cells are in a metabolic steady state, switching to feeding a substrate containing an isotope-labeled substrate. Nutrient solution of the plant, keeping the culture conditions unchanged;

[0048] Start rapid and continuous sampling when the isotope-labeled substrate flows in...

Embodiment 2

[0054] This embodiment provides a method for obtaining the metabolic flux of the intracellular central carbon metabolic pathway under the metabolic stable isotope unstable state, including a batch culture module, a chemostat culture module, a sampling module, and a sample processing and analysis module.

[0055] The concrete steps of described method are as follows:

[0056] Cultivate microorganisms (Saccharomyces cerevisiae CEN.PK113-7D strain) in a 1L bioreactor for batch culture. When the batch culture ends, the oxygen uptake rate (OUR) and CO 2 When the release rate (CER) decreases at the same time, add the nutrient solution without isotope-labeled substrate to the reactor, and start the waste discharge at the same time to carry out chemostat culture;

[0057] When the chemostat culture reaches more than five elution volumes, the microorganisms in the reactor are in a metabolic steady state (OUR and CER both tend to a constant), and the reactor is supplemented with a nutri...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Sizeaaaaaaaaaa
Apertureaaaaaaaaaa
Login to view more

Abstract

The invention discloses a method for obtaining metabolic flux of an intracellular central carbon metabolic pathway in an unsteady state of metabolic steady-state isotopes. The method adopts a batch culture module, a constant culture module, a sampling module and a sample processing and analyzing module. When the OUR and the CER in the batch culture module decline at the same time, culture is performed in the constant culture module. The constant culture module specifically adopts a process of feeding a nutrient solution containing no isotope labeling substrate, starting waste discharge at thesame time, carrying out constant culture, until the microbial cells are in a metabolic steady state, feeding a nutrient solution containing the isotope labeling substrate, and keeping culture conditions unchanged; and when the isotope labeling substrate flows into a reactor, starting rapid and continuous sampling, and entering the sampling module at the same time. According to the method, a seriesof fermentation liquor samples are obtained in a short time, and then the samples are processed and analyzed to obtain accurate intracellular metabolite isotope abundance data; and the method plays acrucial role in calculating the metabolic flux.

Description

technical field [0001] The invention relates to the technical field of calculating metabolic flow, in particular to a method for obtaining the metabolic flux of intracellular central carbon metabolic pathway under the unstable state of metabolic stable isotope. Background technique [0002] The rapid development of "multi-omics" technology has made it possible to obtain the abundance of various biomolecules with high throughput, so that the changes and subtle connections of each omics under different physiological states can be determined. Various omics have been applied in microbial systems biology research, including transcriptomics, which measures mRNA transcription levels; proteomics, which quantifies the abundance of proteins; metabolomics, which measures the abundance of cellular metabolites; metabonomics, which Establishment of flux distribution in intracellular metabolic networks. At the same time, metabolic flux is the result of quantifying the comprehensive networ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/02G01N30/02G01N30/06G01N30/12C12R1/865
CPCC12Q1/02G01N30/02G01N30/06G01N30/12G01N2333/395G01N2030/126
Inventor 夏建业李欢陈敏郑世媛汪佳琪
Owner EAST CHINA UNIV OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap