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Journal of Bacteriology, August 2000, p. 4658-4660, Vol. 182, No. 16
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification, Expression, and Characterization of Escherichia coli Guanine Deaminase

Jason T. Maynes, Richard G. Yuan, and Floyd F. Snyder*

Departments of Medical Genetics and Biochemistry & Molecular Biology, University of Calgary, Calgary, Alberta T2N 4N1, Canada

Received 25 May 2000/Accepted 1 June 2000

Using the human cDNA sequence corresponding to guanine deaminase, the Escherichia coli genome was scanned using the Basic Local Alignment Search Tool (BLAST), and a corresponding 439-residue open reading frame of unknown function was identified as having 36% identity to the human protein. The putative gene was amplified, subcloned into the pMAL-c2 vector, expressed, purified, and characterized enzymatically. The 50.2-kDa protein catalyzed the conversion of guanine to xanthine, having a Km of 15 µM with guanine and a kcat of 3.2 s-1. The bacterial enzyme shares a nine-residue heavy metal binding site with human guanine deaminase, PG[FL]VDTHIH, and was found to contain approximately 1 mol of zinc per mol of subunit of protein. The E. coli guanine deaminase locus is 3' from an open reading frame which shows homology to a bacterial purine base permease.

* Corresponding author. Mailing address: Departments of Medical Genetics and Biochemistry & Molecular Biology, University of Calgary, 3330 Hospital Dr. NW, Calgary, Alberta, Canada T2N 4N1. Phone: (403) 220-6025. Fax: (403) 283-8225. E-mail: snyder{at}ucalgary.ca.

Journal of Bacteriology, August 2000, p. 4658-4660, Vol. 182, No. 16
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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