Applied and Environmental Microbiology, July 2003, p. 3901-3910, Vol. 69, No. 7
0099-2240/03/$08.00+0 DOI: 10.1128/AEM.69.7.3901-3910.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
Identification and Characterization of a GDSL Esterase Gene Located Proximal to the swr Quorum-Sensing System of Serratia liquefaciens MG1
Kathrin Riedel,1 Daniela Talker-Huiber,2 Michael Givskov,3 Helmut Schwab,2 and Leo Eberl1*
Department of Microbiology, Technical University Munich, D-85350 Freising, Germany,1
Department of Biotechnology, Technical University Graz, A-8010 Graz, Austria,2
Department of Microbiology, Technical University of Denmark, DK-2800 Lyngby, Denmark3
Received 26 February 2003/
Accepted 10 April 2003
Serratia liquefaciens MG1 employs the swr quorum-sensing system to control various functions, including production of extracellular enzymes and swarming motility. Here we report the sequencing of the swr flanking DNA regions. We identified a gene upstream of swrR and transcribed in the same direction, designated estA, which encodes an esterase that belongs to family II of lipolytic enzymes. EstA was heterologously expressed in Escherichia coli, and the substrate specificity of the enzyme was determined in crude extracts. With the aid of zymograms visualizing EstA on polyacrylamide gels and by the analysis of a transcriptional fusion of the estA promoter to the promoterless luxAB genes, we showed that expression of the esterase is not regulated by the swr quorum-sensing system. An estA mutant was generated and was found to exhibit growth defects on minimal medium containing Tween 20 or Tween 80 as the sole carbon source. Moreover, we show that the mutant produces greatly reduced amounts of N-acyl-homoserine lactone (AHL) signal molecules on Tween-containing medium compared with the wild type, suggesting that under certain growth conditions EstA may be important for providing the cell with precursors required for AHL biosynthesis.
* Corresponding author. Mailing address: Department of Microbiology, Technical University Munich, Am Hochanger 4, D-85350 Freising, Germany. Phone: 49 8161 715446. Fax: 49 8161 715475. E-mail: eberl{at}mikro.biologie.tu-muenchen.de.
Applied and Environmental Microbiology, July 2003, p. 3901-3910, Vol. 69, No. 7
0099-2240/03/$08.00+0 DOI: 10.1128/AEM.69.7.3901-3910.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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Copyright © 2003 by the American Society for Microbiology. All rights reserved.