Quantification of Polyphosphate in Microalgae by Raman Microscopy and by a Reference Enzymatic Assay

Analytical Chemistry
Šárka MoudříkováPeter Mojzeš

Abstract

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, polyphosphate quantification can be shortened by use of Raman microscopy from days to hours and, additionally, information about intracellular distribution of polyphosphate and heterogeneity among individual cells in algal cult...Continue Reading

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Citations

Aug 20, 2019·The New Phytologist·Laura Lorenzo-OrtsMichael Hothorn
Jul 17, 2020·Frontiers in Plant Science·Emanuel Sanz-LuqueArthur R Grossman
Dec 10, 2020·Proceedings of the National Academy of Sciences of the United States of America·Peter MojzešAnya Salih
Nov 10, 2020·Computational and Structural Biotechnology Journal·Ya-Juan LiuXi-Yong Yu
Jul 21, 2020·Current Opinion in Biotechnology·Dongqi WangApril Z Gu
Jul 3, 2021·International Journal of Molecular Sciences·Simona DzurendováBoris Zimmermann
Feb 11, 2020·Analytical Chemistry·Jonas Johannes ChristLars M Blank

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