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Journal of Bacteriology, March 2004, p. 1565-1570, Vol. 186, No. 5
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.5.1565-1570.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Characterization of Two Methanopterin Biosynthesis Mutants of Methylobacterium extorquens AM1 by Use of a Tetrahydromethanopterin Bioassay

Madeline E. Rasche,^* Stephanie A. Havemann, and Mariana Rosenzvaig

Microbiology and Cell Science Department, University of Florida, Gainesville, Florida 32611-0700

Received 7 July 2003/ Accepted 12 November 2003

An enzymatic assay was developed to measure tetrahydromethanopterin (H₄MPT) levels in wild-type and mutant cells of Methylobacterium extorquens AM1. H₄MPT was detectable in wild-type cells but not in strains with a mutation of either the orf4 or the dmrA gene, suggesting a role for these two genes in H₄MPT biosynthesis. The protein encoded by orf4 catalyzed the reaction of ribofuranosylaminobenzene 5'-phosphate synthase, the first committed step of H₄MPT biosynthesis. These results provide the first biochemical evidence for H₄MPT biosynthesis genes in bacteria.

Florida Agricultural Experiment Station journal series no. R-09891.

Present address: Chemistry Department, University of Florida, Gainesville, Florida.

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