Alkaline phosphatase activity of marine bacteria studied with ELF 97 substrate: Success and limits in the P-limited Mediterranean Sea

France Van Wambeke, Jiří Nedoma, Solange Duhamel, Philippe Lebaron

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

The fluorogenic substrate Enzyme-Labeled Fluorescence 97 (ELF-P) is hydrolyzed by the P-cleaving enzyme phosphatase, producing ELF 97 alcohol (ELFA), a fluorescent-insoluble product. This reaction is used for monitoring phosphatase activity at the single-cell level. Most frequently, ELF-P has been used to determine the P-limitation status of microphytoplankton, but rarely of heterotrophic bacteria. We incubated ELF-P on filters to monitor marine bacterial cultures and oligotrophic Mediterranean Sea samples. Results were compared to classical measurements of bulk alkaline phosphatase activity using the fluorogenic substrate 4-methylumbelliferyl phosphate (MUF-P). A high percentage of the cultured cells were labeled with ELFA (the ratio of ELFA spots to total DAPI counts in P-limited cultures ranged from 26 to 100%, depending on the strain). In contrast, this ratio never exceeded 0.01 % in Mediterranean samples, even when P was demonstrated to be a significant limiting factor. This protocol is useful for application on cruises and with cultures, but was not sufficiently sensitive to detect P-stressed bacterial cells in oligotrophic marine environments.

Original languageEnglish (US)
Pages (from-to)245-251
Number of pages7
JournalAquatic Microbial Ecology
Volume52
Issue number3
DOIs
StatePublished - Sep 2008
Externally publishedYes

Keywords

  • Alkaline phosphatase
  • Cell specific activity
  • ELF97 phosphatase substrate
  • Ectoenzyme
  • Marine bacteria
  • Phosphorus limitation

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Aquatic Science

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