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Applied and Environmental Microbiology, October 2004, p. 6173-6180, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.6173-6180.2004

Quorum Quenching: Enzymatic Disruption of N-Acylhomoserine Lactone-Mediated Bacterial Communication in Burkholderia thailandensis

Ricky L. Ulrich*

Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland

Received 1 April 2004/ Accepted 13 May 2004

Many species of gram-negative bacteria communicate by synthesizing, secreting, and responding to N-acylhomoserine lactones (AHLs), a mechanism termed quorum sensing. Several investigations have characterized numerous AHL-degrading enzymes (AiiA lactonases) encoded by environmental isolates of Bacillus spp. The Burkholderia thailandensis quorum system is comprised of at least three AHL synthases (AHSs) and five transcriptional regulators belonging to the LuxIR class of proteins. Expression of the Bacillus anthracis (Ames strain) AiiA lactonase in B. thailandensis completely abolished the accumulation of N-decanoylhomoserine lactone (C10-HSL) and N-octanoylhomoserine lactone (C8-HSL), reduced N-hexanoylhomoserine lactone (C6-HSL) levels, altered both swarming and twitching motility, caused a significant increase in generation time, and affected carbon metabolism. In contrast, heterologous expression of the Bacillus cereus strain A24 AiiA lactonase in B. thailandensis reduced the concentrations of C6-HSL, C8-HSL, and C10-HSL to nondetectable levels; altered both swarming and twitching motility; and caused fluctuations in carbon utilization. Individual disruption of the B. thailandensis AHSs, specifically disruption of the btaI1 and btaI3 genes, which encode the proteins that direct the synthesis of C8-HSL and C6-HSL, respectively, caused the hyper-beta-hemolysis of sheep erythrocytes on blood agar plates. In contrast, AHL cleavage in B. thailandensis by the Bacillus AiiA lactonases failed to enhance beta-hemolytic activity. The results of this study demonstrate that heterologous expression of Bacillus sp. AiiA lactonases in B. thailandensis reduced AHL accumulation, affected both swarming and twitching motility, increased generation time, altered substrate utilization, and prevented the beta-hemolysis of sheep erythrocytes.

* Mailing address: Bacteriology Division, USAMRIID, 1425 Porter St., Fort Detrick, MD 21702. Phone: (301) 619-8332. Fax: (301) 619-8351. E-mail: Ricky.Ulrich{at}AMEDD.ARMY.MIL.

Applied and Environmental Microbiology, October 2004, p. 6173-6180, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.6173-6180.2004




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