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Carbon-13 nuclear magnetic resonance studies of acetate metabolism in intact cells of Rhodopseudomonas sphaeroides
Authors:Klaas Nicolay  Klaas J Hellingwerf  Robert Kaptein  Wil N Konings
Institution:1. Department of Physical Chemistry, University of Groningen, Nijenborgh 16, 9747 AG Groningen The Netherlands;2. Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN Haren The Netherlands
Abstract:13C-nuclear magnetic resonance was used to study the metabolism of 2-13C]acetate in suspensions of Rhodopseudomonas sphaeroides. In the dark, in logarithmic-phase cells the 13C label appeared first in butyrate C-2 and C-4 and subsequently in glutamate C-4 and succinate C-2 and C-3. In the light, synthesis of poly(β-hydroxybutyrate) (PHB) takes place. Butyrate synthesis seems to be independent of PHB synthesis or degradation activity. Starved, logarithmic-phase cells also show massive synthesis of PHB in the dark. Stationary-phase cells incorporate 13C predominantly into glutamate and succinate. No significant butyrate biosynthesis can be detected in the dark or during illumination. The incorporation of label in PHB is very slow in these cells and most probably originates from exchange of 12C for 13C into PHB. This might indicate slow turnover without net synthesis of the polymer occurring under these conditions. The results are discussed in relation to the redox state and the availability of metabolic energy for biosynthetic reactions in the dark and during illumination of cell suspensions of Rps. sphaeroides.
Keywords:Acetate metabolism  Poly(β-hydroxybutyrate)  Citric acid cycle  (Rps  sphaeroides)  ppm  parts per million  PHB  poly(β-hydroxybutyrate)
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