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J. Bacteriol. doi:10.1128/JB.00337-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Characterization of a novel bile-inducible operon encoding a two component regulatory system in Lactobacillus acidophilus

Genomic Sciences Graduate Program North Carolina State University, Raleigh, NC, 27695, USA; Department of Food Science, North Carolina State University, Raleigh, NC 27695, USA, 27695, USA; Southeast Dairy Foods Research Center, North Carolina State University, Raleigh, NC 27695, USA

^* To whom correspondence should be addressed. Email: trk{at}unity.ncsu.edu.

	Abstract

Lactobacillus acidophilus NCFM is an industrially important strain used extensively as a probiotic culture. Tolerance to the presence of bile is an attribute important to microbial survival in the intestinal tract. A whole genome microarray was employed to examine the effects of bile on the global transcriptional profile of this strain, with the intention of elucidating genes contributing to bile tolerance. Genes involved in carbohydrate metabolism were generally induced, while genes involved in other aspects of cellular growth were mostly repressed. A 7 kb, 8 gene operon encoding a two-component regulatory system, a transporter, an oxidoreductase, and four hypothetical proteins was significantly upregulated in the presence of bile. Deletion mutations were constructed in 6 genes of the operon. Transcriptional analysis of the 2CRS mutants showed that mutation of the HPK had no effect on the induction of the operon, whereas the mutated RR showed enhanced induction when cells were exposed to bile. These results indicate that the 2CRS plays a role in bile tolerance, and the operon it resides in is negatively controlled by the response regulator. Mutations in the transporter, the histidine protein kinase (HPK), the response regulator (RR), and a hypothetical protein each resulted in loss of tolerance to bile. Mutations in genes encoding another hypothetical protein and a putative oxidoreductase resulted in significant increases in bile tolerance. This functional analysis showed that the operon encoded proteins involved in both bile tolerance and bile sensitivity.

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