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Journal of Bacteriology, February 2007, p. 894-901, Vol. 189, No. 3
0021-9193/07/$08.00+0     doi:10.1128/JB.00926-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Anaerobic Central Metabolic Pathways in Shewanella oneidensis MR-1 Reinterpreted in the Light of Isotopic Metabolite Labeling{triangledown}

Yinjie J. Tang,1,2 Adam L. Meadows,2 James Kirby,4 and Jay D. Keasling1,2,3,4*

Synthetic Biology Department, Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California,1 Department of Chemical Engineering, University of California at Berkeley, Berkeley, California,2 Department of Bioengineering, University of California at Berkeley, Berkeley, California,3 California Institute for Quantitative Biomedical Research (QB3), University of California at Berkeley, Berkeley, California4

Received 27 June 2006/ Accepted 5 November 2006

It has been proposed that during growth under anaerobic or oxygen-limited conditions, Shewanella oneidensis MR-1 uses the serine-isocitrate lyase pathway common to many methylotrophic anaerobes, in which formaldehyde produced from pyruvate is condensed with glycine to form serine. The serine is then transformed through hydroxypyruvate and glycerate to enter central metabolism at phosphoglycerate. To examine its use of the serine-isocitrate lyase pathway under anaerobic conditions, we grew S. oneidensis MR-1 on [1-13C]lactate as the sole carbon source, with either trimethylamine N-oxide (TMAO) or fumarate as an electron acceptor. Analysis of cellular metabolites indicated that a large percentage (>70%) of lactate was partially oxidized to either acetate or pyruvate. The 13C isotope distributions in amino acids and other key metabolites indicate that under anaerobic conditions, although glyoxylate synthesized from the isocitrate lyase reaction can be converted to glycine, a complete serine-isocitrate pathway is not present and serine/glycine is, in fact, oxidized via a highly reversible degradation pathway. The labeling data also suggest significant activity in the anapleurotic (malic enzyme and phosphoenolpyruvate carboxylase) reactions. Although the tricarboxylic acid (TCA) cycle is often observed to be incomplete in many other anaerobes (absence of 2-oxoglutarate dehydrogenase activity), isotopic labeling supports the existence of a complete TCA cycle in S. oneidensis MR-1 under certain anaerobic conditions, e.g., TMAO-reducing conditions.

* Corresponding author. Mailing address: Berkeley Center for Synthetic Biology, 717 Potter Street, Building 977, Mail Code 3224, University of California, Berkeley, CA 94720-3224. Phone: (510) 495-2620. Fax: (510) 495-2630. E-mail: keasling{at}berkeley.edu.

{triangledown} Published ahead of print on 17 November 2006.

Journal of Bacteriology, February 2007, p. 894-901, Vol. 189, No. 3
0021-9193/07/$08.00+0     doi:10.1128/JB.00926-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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