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J. Biochem, 1998, Vol. 123, No. 1 115-119
© 1998 Japanese Biochemical Society

research-article

A Two-Dimensional 1H Detected 13C NMR Investigation of Pyruvate Metabolism in Halobacterium salinarium

Ananya Majumdar and Haripalsingh M. Sonawat1

Chemical Physics Group, Tata Institute of Fundamental Research Homi Bhabha Road, Bombay-400 005, India

1To whom correspondence should be addressed. Tel: +91-22-215-2971 (Ext 2271), Fax: +91-22-215-2110/2181, E-mail: hms{at}tifrvax.tifr.res.in

Two-dimensional 1H detected 13C NMR spectroscopy has been used to study the intracellular metabolism of [3–13C]pyruvate in Halobacterium salinarium. The method, resulting in considerable improvement in spectral resolution and signal-to-noise ratio, is well suited for studying transient metabolic intermediates. Pyruvate utilization by the bacterium is a double exponential function with rate constants of 49.13 and 4.67×10–3 per min. The relative 13C enrichment is the fastest for C-3 glutamate. Glutamate C-4 labeling decreases initially and increases later on during incubation, while glutamine C-3 is high to begin with and exhibits a declining trend. The glutamate labeling indicates a high initial flux through pyruvate carboxylase and extensive randomizing of the label in the tricarboxylic acid cycle.


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