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Applied and Environmental Microbiology, October 2005, p. 5794-5804, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.5794-5804.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Microarray Analysis of a Two-Component Regulatory System Involved in Acid Resistance and Proteolytic Activity in Lactobacillus acidophilus

M. Andrea Azcarate-Peril,1 Olivia McAuliffe,1,{dagger} Eric Altermann,1 Sonja Lick,1,{ddagger} W. Michael Russell,2,§ and Todd R. Klaenhammer1*

Department of Food Science, North Carolina State University, Raleigh, North Carolina,1 Danisco, Inc., Madison, Wisconsin2

Received 17 December 2004/ Accepted 13 May 2005

Two-component regulatory systems are one primary mechanism for environmental sensing and signal transduction. Annotation of the complete genome sequence of the probiotic bacterium Lactobacillus acidophilus NCFM revealed nine two-component regulatory systems. In this study, the histidine protein kinase of a two-component regulatory system (LBA1524HPK-LBA1525RR), similar to the acid-related system lisRK from Listeria monocytogenes (P. D. Cotter et al., J. Bacteriol. 181:6840-6843, 1999), was insertionally inactivated. A whole-genome microarray containing 97.4% of the annotated genes of L. acidophilus was used to compare genome-wide patterns of transcription at various pHs between the control and the histidine protein kinase mutant. The expression pattern of approximately 80 genes was affected by the LBA1524HPK mutation. Putative LBA1525RR target loci included two oligopeptide-transport systems present in the L. acidophilus genome, other components of the proteolytic system, and a LuxS homolog, suspected of participating in synthesis of the AI-2 signaling compound. The mutant exhibited lower tolerance to acid and ethanol in logarithmic-phase cells and poor acidification rates in milk. Supplementation of milk with Casamino Acids essentially restored the acid-producing ability of the mutant, providing additional evidence for a role of this two component system in regulating proteolytic activity in L. acidophilus.

* Corresponding author. Mailing address: Department of Food Science, North Carolina State University, Box 7624, Raleigh, NC 27695. Phone: (919) 515-2972. Fax: (919) 515-7124. E-mail: klaenhammer{at}ncsu.edu.

{dagger} Present address: Dairy Products Research Centre, Teagasc, Moorepark, Fermoy, County Cork, Ireland.

{ddagger} Present address: Bundesinstitut für Risikobewertung, Standort Berlin-Marienfelde, Diedersdorfer Weg 1, D-12277 Berlin, Germany.

§ Present address: Mead Johnson Nutritionals, 2400 W. Lloyd Expressway, Evansville, IN 47721.

Applied and Environmental Microbiology, October 2005, p. 5794-5804, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.5794-5804.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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