Moudrikova, Sarka, Sadowsky, Andres, Metzger, Sabine, Nedbal, Ladislav, Mettler-Altmann, Tabea and Mojzes, Peter ORCID: 0000-0002-9952-6939 (2017). Quantification of Polyphosphate in Microalgae by Raman Microscopy and by a Reference Enzymatic Assay. Anal. Chem., 89 (22). S. 12006 - 12014. WASHINGTON: AMER CHEMICAL SOC. ISSN 1520-6882

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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 R-2 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 culture can be obtained. This offers new insights into the dynamics and role of these polymers crucial for phosphorus uptake and storage. This analytical capability is of particular practical importance because algae aid phosphorus sequestration from wastewater and the thus enriched biomass may serve as organic fertilizer. Both these applications have a strong potential in a future sustainable, circular bioeconomy.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Moudrikova, SarkaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Sadowsky, AndresUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Metzger, SabineUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Nedbal, LadislavUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Mettler-Altmann, TabeaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Mojzes, PeterUNSPECIFIEDorcid.org/0000-0002-9952-6939UNSPECIFIED
URN: urn:nbn:de:hbz:38-211018
DOI: 10.1021/acs.analchem.7b02393
Journal or Publication Title: Anal. Chem.
Volume: 89
Number: 22
Page Range: S. 12006 - 12014
Date: 2017
Publisher: AMER CHEMICAL SOC
Place of Publication: WASHINGTON
ISSN: 1520-6882
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
ALGA DUNALIELLA-SALINA; INORGANIC POLYPHOSPHATE; CHLAMYDOMONAS-REINHARDTII; SACCHAROMYCES-CEREVISIAE; CHLORELLA-VULGARIS; PHOSPHATE-UPTAKE; METABOLISM; GROWTH; SURVIVAL; PHOTOSYNTHESISMultiple languages
Chemistry, AnalyticalMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/21101

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