A nonradioactive method for the assay of polyphosphate kinase activity and its application in the study of polyphosphate metabolism in Burkholderia cepacia

A. Mullan, John Quinn, John McGrath

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

Studies of polyphosphate (polyP) metabolism in microorganisms have been hampered by the lack of a convenient method for the assay in cell extracts of the activity of polyphosphate kinase (PPK), the enzyme principally responsible for microbial polyP biosynthesis. We report the development of such an assay, based on the well-established metachromatic reaction, with toluidine blue, of the polyP formed during the PPK-catalyzed reaction. The method was successfully used in the characterization of PPK activity in crude extracts of an environmental Burkholderia cepacia isolate. The development of a protocol for the physical recovery of polyP from solution is also reported. (C) 2002 Elsevier Science (USA). All rights reserved.
Original languageEnglish
Pages (from-to)294-299
Number of pages6
JournalAnalytical Biochemistry
Volume308
Issue number2
Publication statusPublished - 15 Oct 2002

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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