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Improvement of biochemical methods of polyP quantification

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dc.contributor.author Bru, Samuel
dc.contributor.author Jiménez, Javier
dc.contributor.author Canadell, David
dc.contributor.author Ariño, Joaquín
dc.contributor.author Clotet Erra, Josep
dc.date.accessioned 2023-11-20T08:25:58Z
dc.date.available 2023-11-20T08:25:58Z
dc.date.issued 2017
dc.identifier.citation Bru S, Jimenez J, Canadell D, Arino J, Clotet J. Improvement of biochemical methods of polyP quantification. Microb Cell. 2017 Jan;4(1):6-15. DOI: 10.15698/mic2017.01.551
dc.identifier.issn 2311-2638
dc.identifier.uri http://hdl.handle.net/10230/58313
dc.description.abstract Polyphosphate (polyP) is an abundant and physiologically important biomolecule for virtually any living cell. Therefore, determination of changes in cellular content of polyP is crucial for its functional characterization. Determination of cellular polyP has been performed by many different methods, and the lack of a standardized procedure is possibly responsible for the large dispersion of results found in the relevant literature. For a relatively simple organism, such as the yeast Saccharomyces cerevisiae, this variation can be up to 12-fold. polyP extraction and determination of free phosphate released by enzymatic degradation of the polymer is a method quite common and relatively straightforward for polyP determination. By using the yeast S. cerevisiae as model, we have experimentally evaluated the different steps in this procedure in order to identify critical issues that might explain the disparate reported results. As the main output of this evaluation we propose a straightforward and robust procedure that can be used as gold standard protocol for cellular polyP purification and determination from unicellular organisms, thus providing consistency to measurements and facilitating inter-laboratory comparisons and biological interpretation of the results.
dc.description.sponsorship We want to thanks A. Kornberg for the plasmid pTrcPPX1 and M. Pérez for invaluable technical assistance. This work has been funded by the Spanish Ministerio de Economía y Competitividad MINECO grants ref: BFU 2013-44189-P to JC ; BFU2011-30197-C3-01 and BFU2014-54591-C2-1-P to JA. SB was the recipient of a grant from the UIC.
dc.format.mimetype application/pdf
dc.language.iso eng
dc.publisher Shared Science Publishers
dc.relation.ispartof Microbial Cell. 2017 Jan;4(1):6-15
dc.rights © 2016 Bru et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduction in any medium, provided the original author and source are acknowledged.
dc.rights.uri http://creativecommons.org/licenses/by/4.0/
dc.title Improvement of biochemical methods of polyP quantification
dc.type info:eu-repo/semantics/article
dc.identifier.doi http://dx.doi.org/10.15698/mic2017.01.551
dc.subject.keyword Polyphosphate
dc.subject.keyword Yeast
dc.subject.keyword Saccharomyces cerevisiae
dc.subject.keyword Neutral-phenol
dc.relation.projectID info:eu-repo/grantAgreement/ES/1PE/BFU 2013-44189-P
dc.relation.projectID info:eu-repo/grantAgreement/ES/1PE/BFU2014-54591-C2-1-P
dc.relation.projectID info:eu-repo/grantAgreement/ES/3PN/BFU2011-30197-C3-01
dc.rights.accessRights info:eu-repo/semantics/openAccess
dc.type.version info:eu-repo/semantics/publishedVersion


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