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Characterization of the codY gene and its influence on biofilm formation in Bacillus cereus

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Abstract

The foodborne pathogen Bacillus cereus can form biofilms on various food contact surfaces, leading to contamination of food products. To study the mechanisms of biofilm formation by B. cereus, a Tn5401 library was generated from strain UW101C. Eight thousand mutants were screened in EPS, a low nutrient medium. One mutant (M124), with a disruption in codY, developed fourfold less biofilm than the wild-type, and its defective biofilm phenotype was rescued by complementation. Addition of 0.1% casamino acids to EPS prolonged the duration of biofilms in the wild-type but not codY mutant. When decoyinine, a GTP synthesis inhibitor, was added to EPS, biofilm formation was decreased in the wild-type but not the mutant. The codY mutant produced three times higher protease activity than the wild-type. Zymogram and SDS-PAGE data showed that production of the protease (∼130 kDa) was repressed by CodY. Addition of proteinase K to EPS decreased biofilm formation by the wild-type. Using a dpp-lacZ fusion reporter system, it was shown that that the B. cereus CodY can sense amino acids and GTP levels. These data suggest that by responding to amino acids and intracellular GTP levels CodY represses production of an unknown protease and is involved in biofilm formation.

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Acknowledgments

This research was supported by Hatch funds (WIS04799) and by the College of Agricultural and Life Sciences, University of Wisconsin-Madison. We are grateful to J. Handelsman for supplying B. cereus UW101C and pEG922, D. Lereclus for pHT304, and A. L. Sonenshein for B. subtilis PS59 and PS164.

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Author notes

  1. Yi-Huang Hsueh

    Present address: Department of Biology, Indiana University, 1001 East Third Street, Bloomington, IN, 47405, USA

Authors and Affiliations

  1. Department of Bacteriology, Food Research Institute, University of Wisconsin, 1550 Linden Drive, Madison, WI, 53706, USA

    Yi-Huang Hsueh, Eileen B. Somers & Amy C. Lee Wong

Authors
  1. Yi-Huang Hsueh
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  2. Eileen B. Somers
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  3. Amy C. Lee Wong
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Correspondence to Amy C. Lee Wong.

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Communicated by Erko Stackebrandt.

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Hsueh, YH., Somers, E.B. & Wong, A.C.L. Characterization of the codY gene and its influence on biofilm formation in Bacillus cereus . Arch Microbiol 189, 557–568 (2008). https://doi.org/10.1007/s00203-008-0348-8

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  • DOI: https://doi.org/10.1007/s00203-008-0348-8

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