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Applied and Environmental Microbiology, January 2003, p. 504-508, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.504-508.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Gene Cloning, Purification, and Characterization of a Phosphodiesterase from Delftia acidovorans

Sundiep K. Tehara and Jay D. Keasling*

Department of Chemical Engineering, University of California at Berkeley, Berkeley, California 94720-1462

Received 17 June 2002/ Accepted 7 October 2002

A novel phosphodiesterase (PdeA) was purified from Delftia acidovorans, the gene encoding the enzyme was cloned and expressed in Escherichia coli, and the recombinant enzyme was purified to apparent homogeneity and characterized. PdeA is an 85-kDa trimer that exhibits maximal activity at 65°C and pH 10 even though it was isolated from a mesophilic bacterium. Although PdeA exhibited both mono- and diesterase activity, it was most active on the phosphodiester bis(p-nitrophenyl)phosphate with a Km of 2.9 ± 0.1 mM and a kcat of 879 ± 73 min-1. The enzyme showed sequence similarity to cyclic AMP (cAMP) phosphodiesterase and cyclic nucleotide phosphodiesterases and exhibited activity on cAMP in vivo when the gene was expressed in E. coli. The IS1071 transposon insertion sequence was found downstream of pdeA.

* Corresponding author. Mailing address: Department of Chemical Engineering, University of California at Berkeley, Berkeley, CA 94720-1462. Phone: (510) 642-4862. Fax: (510) 643-1228. E-mail: keasling{at}socrates.berkeley.edu.

Applied and Environmental Microbiology, January 2003, p. 504-508, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.504-508.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • de la Pena Mattozzi, M., Tehara, S. K., Hong, T., Keasling, J. D. (2006). Mineralization of Paraoxon and Its Use as a Sole C and P Source by a Rationally Designed Catabolic Pathway in Pseudomonas putida.. Appl. Environ. Microbiol. 72: 6699-6706 [Abstract] [Full Text]  
  • McLoughlin, S. Y., Jackson, C., Liu, J.-W., Ollis, D. L. (2004). Growth of Escherichia coli Coexpressing Phosphotriesterase and Glycerophosphodiester Phosphodiesterase, Using Paraoxon as the Sole Phosphorus Source. Appl. Environ. Microbiol. 70: 404-412 [Abstract] [Full Text]  


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