Journal of Bacteriology, January 1999, p. 389-395, Vol. 181, No. 2
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
School of Animal and Microbial Sciences,
Received 3 August 1998/Accepted 25 September 1998
In Escherichia coli, the phn operon encodes
proteins responsible for the uptake and breakdown of phosphonates. The
C-P (carbon-phosphorus) lyase enzyme encoded by this operon which
catalyzes the cleavage of C-P bonds in phosphonates has been
recalcitrant to biochemical characterization. To advance the
understanding of this enzyme, we have cloned DNA from
Rhizobium (Sinorhizobium) meliloti
that contains homologues of the E. coli phnG,
-H, -I, -J, and -K
genes. We demonstrated by insertional mutagenesis that the operon from which this DNA is derived encodes the R. meliloti C-P
lyase. Furthermore, the phenotype of this phn mutant shows
that the C-P lyase has a broad substrate specificity and that the
organism has another enzyme that degrades aminoethylphosphonate. A
comparison of the R. meliloti and E. coli phn
genes and their predicted products gave new information about C-P
lyase. The putative R. meliloti PhnG, PhnH, and PhnK
proteins were overexpressed and used to make polyclonal antibodies.
Proteins of the correct molecular weight that react with these
antibodies are expressed by R. meliloti grown with
phosphonates as sole phosphorus sources. This is the first in vivo
demonstration of the existence of these hitherto hypothetical Phn proteins.
*
Corresponding author. Mailing address: School of Animal
and Microbial Sciences, University of Reading, Whiteknights, P.O. Box
228, Reading RG6 6AJ, United Kingdom. Phone: (44) 1189 316639. Fax:
(44) 1189 316562. E-mail: sksrobsn{at}reading.ac.uk.
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