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Journal of Bacteriology, March 2002, p. 1750-1758, Vol. 184, No. 6
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.6.1750-1758.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Glyoxylate Regeneration Pathway in the Methylotroph Methylobacterium extorquens AM1{dagger}

Natalia Korotkova,1 Ludmila Chistoserdova,1 Vladimir Kuksa,2 and Mary E. Lidstrom1,3*

Department of Chemical Engineering,1 Department of Ophthalmology,2 Department of Microbiology, University of Washington, Seattle, Washington 98195-17503

Received 26 September 2001/ Accepted 30 November 2001

Most serine cycle methylotrophic bacteria lack isocitrate lyase and convert acetyl coenzyme A (acetyl-CoA) to glyoxylate via a novel pathway thought to involve butyryl-CoA and propionyl-CoA as intermediates. In this study we have used a genome analysis approach followed by mutation to test a number of genes for involvement in this novel pathway. We show that methylmalonyl-CoA mutase, an R-specific crotonase, isobutyryl-CoA dehydrogenase, and a GTPase are involved in glyoxylate regeneration. We also monitored the fate of 14C-labeled carbon originating from acetate, butyrate, or bicarbonate in mutants defective in glyoxylate regeneration and identified new potential intermediates in the pathway: ethylmalonyl-CoA, methylsuccinyl-CoA, isobutyryl-CoA, methacrylyl-CoA, and ß-hydroxyisobutyryl-CoA. A new scheme for the pathway is proposed based on these data.

* Corresponding author. Mailing address: Department of Chemical Engineering, University of Washington, Seattle, WA 98195-1750. Phone: (206) 616-5282. Fax: (206) 616-5721. E-mail: lidstrom{at}u.washington.edu.

{dagger} Dedicated to J. R. Quayle.

Journal of Bacteriology, March 2002, p. 1750-1758, Vol. 184, No. 6
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.6.1750-1758.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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