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Journal of Bacteriology, April 2000, p. 2113-2118, Vol. 182, No. 8
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Requirement for the Enzymes Acetoacetyl Coenzyme A Synthetase and Poly-3-Hydroxybutyrate (PHB) Synthase for Growth of Sinorhizobium meliloti on PHB Cycle Intermediates

Guo-qin Cai,1 Brian T. Driscoll,1 and Trevor C. Charles1,2,*

Department of Natural Resource Sciences, McGill University, Ste.-Anne-de-Bellevue, Québec H9X 3V9,1 and Department of Biology, University of Waterloo, Waterloo, Ontario N2L 3G1,2 Canada

Received 22 October 1999/Accepted 22 January 2000

We have identified two Sinorhizobium meliloti chromosomal loci affecting the poly-3-hydroxybutyrate degradation pathway. One locus was identified as the gene acsA, encoding acetoacetyl coenzyme A (acetoacetyl-CoA) synthetase. Analysis of the acsA nucleotide sequence revealed that this gene encodes a putative protein with a molecular weight of 72,000 that shows similarity to acetyl-CoA synthetase in other organisms. Acetyl-CoA synthetase activity was not affected in cell extracts of glucose-grown acsA::Tn5 mutants; instead, acetoacetyl-CoA synthetase activity was drastically reduced. These findings suggest that acetoacetyl-CoA synthetase, rather than CoA transferase, activates acetoacetate to acetoacetyl-CoA in the S. meliloti poly-3-hydroxybutyrate cycle. The second locus was identified as phbC, encoding poly-3-hydroxybutyrate synthase, and was found to be required for synthesis of poly-3-hydroxybutyrate deposits.

* Corresponding author. Mailing address: Department of Biology, University of Waterloo, 200 University Ave. West, Waterloo, ON N2L 3G1, Canada. Phone: (519) 888-4567, ext. 5606. Fax: (519) 746-0614. E-mail: tcharles{at}uwaterloo.ca.

Journal of Bacteriology, April 2000, p. 2113-2118, Vol. 182, No. 8
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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