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Applied and Environmental Microbiology, July 2002, p. 3486-3495, Vol. 68, No. 7
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.7.3486-3495.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of Proteins Involved in the Heat Stress Response of Bacillus cereus ATCC 14579

Paula M. Periago,1,2,{dagger} Willem van Schaik,1,2 Tjakko Abee,1,2* and Jeroen A. Wouters1,2,{ddagger}

Wageningen Center for Food Sciences,1 Laboratory of Food Microbiology, Wageningen University, 6703 HD Wageningen, The Netherlands2

Received 13 December 2001/ Accepted 19 April 2002

To monitor the ability of the food-borne opportunistic pathogen Bacillus cereus to survive during minimal processing of food products, we determined its heat-adaptive response. During pre-exposure to 42°C, B. cereus ATCC 14579 adapts to heat exposure at the lethal temperature of 50°C (maximum protection occurs after 15 min to 1 h of pre-exposure to 42°C). For this heat-adaptive response, de novo protein synthesis is required. By using two-dimensional gel electrophoresis, we observed 31 heat-induced proteins, and we determined the N-terminal sequences of a subset of these proteins. This revealed induction of stress proteins (CspB, CspE, and SodA), proteins involved in sporulation (SpoVG and AldA), metabolic enzymes (FolD and Dra), identified heat-induced proteins in related organisms (DnaK, GroEL, ClpP, RsbV, HSP16.4, YflT, PpiB, and TrxA), and other proteins (MreB, YloH, and YbbT). The upregulation of several stress proteins was confirmed by using antibodies specific for well-characterized heat shock proteins (HSPs) of B. subtilis. These observations indicate that heat adaptation of B. cereus involves proteins that function in a variety of cellular processes. Notably, a 30-min pre-exposure to 4% ethanol, pH 5, or 2.5% NaCl also results in increased thermotolerance. Also, for these adaptation processes, protein synthesis is required, and indeed, some HSPs are induced under these conditions. Collectively, these data show that during mild processing, cross-protection from heating occurs in pathogenic B. cereus, which may result in increased survival in foods.

* Corresponding author. Mailing address: Laboratory of Food Microbiology, Wageningen University, Bomenweg 2, 6703 HD Wageningen, The Netherlands. Phone: 31-317-484981. Fax: 31-317-484978. E-mail: Tjakko.Abee{at}micro.fdsci.wau.nl.

{dagger} Present address: Universidad Politécnica de Cartagena, Dpto. Ingeniería de Alimentos y del Equipamiento Agrícola, 30203 Cartagena, Spain.

{ddagger} Present address: Department Flavor, Nutrition and Ingredients, NIZO Food Research, 6710 BA Ede, The Netherlands.

Applied and Environmental Microbiology, July 2002, p. 3486-3495, Vol. 68, No. 7
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.7.3486-3495.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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