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J. Biol. Chem., Vol. 280, Issue 17, 16748-16753, April 29, 2005

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Genetically Engineered Production of 1-Desmethylcobyrinic Acid, 1-Desmethylcobyrinic Acid a,c-Diamide, and Cobyrinic Acid a,c-Diamide in Escherichia coli Implies a Role for CbiD in C-1 Methylation in the Anaerobic Pathway to Cobalamin^*

Charles A. Roessner, Howard J. Williams, and A. Ian Scott

From the Center for Biological NMR, Department of Chemistry, Texas A&M University, College Station, Texas 77843-3255

Co-expression of the cobA gene from Propionibacterium freudenreichii and the cbiA, -C, -D, -E, -T, -F, -G, -H, -J, -K, -L, and -P genes from Salmonella enterica serovar typhimurium in Escherichia coli resulted in the production of cobyrinic acid a,c-diamide. A cbiD deletion mutant of this strain produced 1-desmethylcobyrinic acid a,c-diamide, indicating that CbiD is involved in C-1 methylation in the anaerobic pathway to cobalamin. Strains that did not have the cbiP gene also produced 1-desmethylcobyrinic acid a,c-diamide, and strains that had neither cbiP nor cbiA synthesized 1-desmethylcobyrinic acid even in the presence of cbiD, suggesting that CbiA and CbiP are necessary for CbiD activity.

Received for publication, February 17, 2005

^* This work was supported by National Institutes of Health MERIT Award DK32034 (to A. I. S.) and the Robert A. Welch Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental material.

To whom correspondence should be addressed. Tel.: 979-845-8985; Fax: 979-845-5992; E-Mail: c-roessner{at}tamu.edu.

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