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Glyoxylate Regeneration Pathway in the Methylotroph Methylobacterium extorquens AM1

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.

Dedicated to J. R. Quayle.

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