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Metabolites of the Free-Living Amoeba Phreatamoeba balamuthi Analyzed by 13C- and 31P-NMR Spectroscopy: Occurrence of Phosphoinositol Diphosphates
Authors:JEAN-BAPTISTE MARTIN  TILLY BAKKER-GRUNWALD  GERARD KLEIN
Affiliation:Laboratoire de Résonance Magnétique en Biologie et Médecine, Département de Biologie Moléculaire et Structurale, CEA-Centre d'Etudes Nucléaires de Grenoble, France;Abteilung Mikrobiologie, Universität Osnabrück, Germany;Laboratoire de Biologie Cellulaire (URA 1130 du CNRS), Département de Biologie Moléculaire et Structurale, CEA-Centre d'Etudes Nucléaires de Grenoble, France
Abstract:ABSTRACT. Phreatamoeba balamuthi is a free-living heterotrophic amoeba that lacks mitochondria. Metabolites of axenically-grown cells were characterized by natural-abundance 13C-NMR and 31P-NMR spectroscopy on acellular perchloric acid extracts. The amoebae were found to contain glycogen and trehalose as storage carbohydrates, together with putrescine and several amino acids, most prominently proline; we propose that proline and trehalose may serve in osmoregulation. Glycerophosphocholine and glycerophosphoethanolamine were present with their phosphomonoester derivatives, phosphocholine and phosphoethanolamine. Along with inorganic phosphate, inorganic pyrophosphate, nucleoside diphosphates, nucleoside triphosphates and NAD, P. balamuthi amoebae also contained unusual phosphoinositol diphosphates in large quantities (0.5 μmol/g wet cells).
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