Journal article
RapidRIP quantifies the intracellular metabolome of 7 industrial strains of E. coli
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Network Reconstruction in Silico Biology, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
iLoop, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
University of California at San Diego4
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
Global Econometric Modeling, Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark6
Fast metabolite quantification methods are required for high throughput screening of microbial strains obtained by combinatorial or evolutionary engineering approaches. In this study, a rapid RIP-LC-MS/MS (RapidRIP) method for high-throughput quantitative metabolomics was developed and validated that was capable of quantifying 102 metabolites from central, amino acid, energy, nucleotide, and cofactor metabolism in less than 5 minutes.
The method was shown to have comparable sensitivity and resolving capability as compared to a full length RIP-LC-MS/MS method (FullRIP). The RapidRIP method was used to quantify the metabolome of seven industrial strains of E. coli revealing significant differences in glycolytic, pentose phosphate, TCA cycle, amino acid, and energy and cofactor metabolites were found.
These differences translated to statistically and biologically significant differences in thermodynamics of biochemical reactions between strains that could have implications when choosing a host for bioprocessing.
Language: | English |
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Year: | 2018 |
Pages: | 383-392 |
ISSN: | 10967184 and 10967176 |
Types: | Journal article |
DOI: | 10.1016/j.ymben.2018.04.009 |
ORCIDs: | Schrübbers, Lars , Christensen, Hanne B. and Herrgård, Markus J. |