Quantitative determination of intracellular adenine nucleotides and coenzymes from bacterium Paracoccus denitrificans toward to targeted metabolome analysis
| Authors | |
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| Year of publication | 2008 |
| Type | Article in Proceedings |
| Conference | Book of Abstracts CECE 2008 |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | Target metabolome analysis; metabolite extraction |
| Description | Metabolomics (metabolome analysis) is comprehensive analysis of small molecules (metabolites) in the cell. Because of the large differences in abundance and wide diversity in physicochemical properties of metabolites in organism the metabolome analysis is a difficult analytical task. So several approaches of metabolomics studies are known; e.g. the targeted metabolome analysis is focused only on quantitation of specific metabolites or metabolite classes. The development of robust a consistent experimental protocol for all steps in the procedure ranging from biomass cultivation, metabolism quenching and metabolite extraction to the quantitative analysis is required. The main aim of this study was to optimize procedure for isolation and concentration of targeted metabolites from the biological matrix of bacterium Paracoccus denitrificans with the least possible losses. Organic solvents like methanol, ethanol, acetonitrile and others are often used for extraction of intracellular metabolites from bacterial cells. In this consequence these extraction media were tested: 20, 50, 80 and 100 % methanol, ethanol, acetonitrile and furthermore mixture of acetonitrile:methanol:water (40:40:20), mixture of acidic (0.1 M formic acid containing) organic solvents and mixtures of basic (0.1 M NH4OH containing) organic solvents. Metabolite yields of individual extractions were measured by already optimized CZE method with UV detection and compared each other. |
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