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Journal of Bacteriology, June 2006, p. 4227-4235, Vol. 188, No. 12
0021-9193/06/$08.00+0     doi:10.1128/JB.00227-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The cbiS Gene of the Archaeon Methanopyrus kandleri AV19 Encodes a Bifunctional Enzyme with Adenosylcobinamide Amidohydrolase and {alpha}-Ribazole-Phosphate Phosphatase Activities

Jesse D. Woodson{dagger} and Jorge C. Escalante-Semerena*

Department of Bacteriology, University of Wisconsin—Madison, Madison, Wisconsin

Received 13 February 2006/ Accepted 5 April 2006

Here we report the initial biochemical characterization of the bifunctional {alpha}-ribazole-P ({alpha}-RP) phosphatase, adenosylcobinamide (AdoCbi) amidohydrolase CbiS enzyme from the hyperthermophilic methanogenic archaeon Methanopyrus kandleri AV19. The cbiS gene encodes a 39-kDa protein with two distinct segments, one of which is homologous to the AdoCbi amidohydrolase (CbiZ, EC 3.5.1.90) enzyme and the other of which is homologous to the recently discovered archaeal {alpha}-RP phosphatase (CobZ, EC 3.1.3.73) enzyme. CbiS function restored AdoCbi salvaging and {alpha}-RP phosphatase activity in strains of the bacterium Salmonella enterica where either step was blocked. The two halves of the cbiS genes retained their function in vivo when they were cloned separately. The CbiS enzyme was overproduced in Escherichia coli and was isolated to >95% homogeneity. High-performance liquid chromatography, UV-visible spectroscopy, and mass spectroscopy established {alpha}-ribazole and cobyric acid as the products of the phosphatase and amidohydrolase reactions, respectively. Reasons why the CbiZ and CobZ enzymes are fused in some archaea are discussed.

* Corresponding author. Mailing address: 144A Enzyme Institute, 1710 University Avenue, Madison, WI 53726-4087. Phone: (608) 262-7379. Fax: (608) 265-7909. E-mail: escalante{at}bact.wisc.edu.

{dagger} Present address: The Salk Institute Plant Biology Laboratory, La Jolla, CA 92037.

Journal of Bacteriology, June 2006, p. 4227-4235, Vol. 188, No. 12
0021-9193/06/$08.00+0     doi:10.1128/JB.00227-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gray, M. J., Escalante-Semerena, J. C. (2009). In Vivo Analysis of Cobinamide Salvaging in Rhodobacter sphaeroides Strain 2.4.1. J. Bacteriol. 191: 3842-3851 [Abstract] [Full Text]  
  • Otte, M. M., Woodson, J. D., Escalante-Semerena, J. C. (2007). The Thiamine Kinase (YcfN) Enzyme Plays a Minor but Significant Role in Cobinamide Salvaging in Salmonella enterica. J. Bacteriol. 189: 7310-7315 [Abstract] [Full Text]  
  • Escalante-Semerena, J. C. (2007). Conversion of Cobinamide into Adenosylcobamide in Bacteria and Archaea. J. Bacteriol. 189: 4555-4560 [Full Text]  


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