Quantification of Polyphosphate in Microalgae by Raman Microscopy and by a Reference Enzymatic Assay
| Autor: | Tabea Mettler-Altmann, Ladislav Nedbal, Peter Mojzeš, Šárka Moudříková, Sabine Metzger, Andres Sadowsky |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Microorganism Chlorella vulgaris Saccharomyces cerevisiae Wastewater Spectrum Analysis Raman 01 natural sciences Analytical Chemistry 03 medical and health sciences symbols.namesake chemistry.chemical_compound Polyphosphates Microscopy Ethanol precipitation chemistry.chemical_classification Chemistry Polyphosphate Assay Acid Anhydride Hydrolases 030104 developmental biology Enzyme Biochemistry symbols Raman spectroscopy 010606 plant biology & botany |
| Zdroj: | Analytical Chemistry. 89:12006-12013 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.7b02393 |
| Popis: | Polyphosphates have occurred in living cells early in evolution and microalgae contain these important polymers in their cells. Progress in research of polyphosphate metabolism of these ecologically as well as biotechnologically important microorganisms is hampered by the lack of rapid quantification methods. Experiments with the green alga Chlorella vulgaris presented here compared polyphosphate extraction in water, methanol-chloroform, and phenol-chloroform followed by polyphosphate purification by binding to silica columns or ethanol precipitation. The phenol-chloroform extraction of C. vulgaris followed by ethanol precipitation of polyphosphate was shown to be superior to the other tested method variants. Recovery test of added polyphosphate standard to algal biomass showed that the method is accurate. Using this biochemical assay as a validated reference, we show that 2-dimensional, confocal Raman microscopy can serve as a linear proxy for polyphosphate in C. vulgaris with R2 up to 0.956. With this, ... |
| Databáze: | OpenAIRE |
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