AME 52:245-251 (2008)  -  DOI: https://doi.org/10.3354/ame01238

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

France Van Wambeke1,*, Jirí Nedoma2, Solange Duhamel1, Philippe Lebaron3,4

1Laboratoire de Microbiologie, Géochimie et Ecologie Marines, CNRS, UMR 6117, Case 901, Centre d’Océanologie de Marseille, Université de la Méditerranée - Campus de Luminy, 13288 Marseille Cedex 9, France
2Biological Centre of the Academy of Sciences of the Czech Republic, Hydrobiological Institute, Na sádkách 7,
37005 Ceské Budejovice, Czech Republic
3Université Pierre et Marie Curie-Paris 6, and 4CNRS, UMR7621, 66650 Banyuls-sur-Mer, France

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.


KEY WORDS: Marine bacteria · Alkaline phosphatase · Cell specific activity · Ectoenzyme · ELF 97 phosphatase substrate · Phosphorus limitation


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Cite this article as: Van Wambeke F, Nedoma J, Duhamel S, Lebaron P (2008) Alkaline phosphatase activity of marine bacteria studied with ELF 97 substrate: success and limits in the P-limited Mediterranean Sea. Aquat Microb Ecol 52:245-251. https://doi.org/10.3354/ame01238

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