RapidRIP quantifies the intracellular metabolome of 7 industrial strains of E. coli
| Autor: | Douglas McCloskey, Julia Xu, Markus J. Herrgård, Hanne Bjerre Christensen, Lars Schrübbers |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
High-throughput screening Metabolite Bioengineering Pentose phosphate pathway Applied Microbiology and Biotechnology Mass Spectrometry Cofactor 03 medical and health sciences chemistry.chemical_compound Metabolomics Species Specificity Escherichia coli Metabolome chemistry.chemical_classification 030102 biochemistry & molecular biology biology Amino acid Citric acid cycle 030104 developmental biology chemistry Biochemistry biology.protein Chromatography Liquid Biotechnology |
| Zdroj: | McCloskey, D, Xu, J, Schrübbers, L, Christensen, H B & Herrgård, M J 2018, ' RapidRIP quantifies the intracellular metabolome of 7 industrial strains of E. coli ', Metabolic Engineering, vol. 47, pp. 383-392 . https://doi.org/10.1016/j.ymben.2018.04.009 Metabolic Engineering |
| ISSN: | 1096-7176 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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