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Journal of Bacteriology, May 2008, p. 3161-3168, Vol. 190, No. 9
0021-9193/08/$08.00+0     doi:10.1128/JB.01637-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Identification of Genes Associated with the Long-Gut-Persistence Phenotype of the Probiotic Lactobacillus johnsonii Strain NCC533 Using a Combination of Genomics and Transcriptome Analysis{triangledown} ,{dagger}

Emmanuel Denou,1,2 Raymond David Pridmore,1 Bernard Berger,1 Jean-Michel Panoff,2 Fabrizio Arigoni,1 and Harald Brüssow1*

Nestlé Research Center, Nestec Ltd., P.O. Box 44, CH-1000 Lausanne 26, Switzerland,1 Laboratoire de Microbiologie Alimentaire, IBFA-ISBIO, Université de Caen-Basse Normandie, F-14032 Caen cedex, France2

Received 9 October 2007/ Accepted 17 January 2008

Lactobacillus johnsonii strains NCC533 and ATCC 33200 (the type strain of this species) differed significantly in gut residence time (12 versus 5 days) after oral feeding to mice. Genes affecting the long gut residence time of the probiotic strain NCC533 were targeted for analysis. We hypothesized that genes specific for this strain, which are expressed during passage of the bacterium through the gut, affect the phenotype. When the DNA of the type strain was hybridized against a microarray of the sequenced NCC533 strain, we identified 233 genes that were specific for the long-gut-persistence isolate. Whole-genome transcription analysis of the NCC533 strain using the microarray format identified 174 genes that were strongly and consistently expressed in the jejunum of mice monocolonized with this strain. Fusion of the two microarray data sets identified three gene loci that were both expressed in vivo and specific to the long-gut-persistence isolate. The identified genes included LJ1027 and LJ1028, two glycosyltransferase genes in the exopolysaccharide synthesis operon; LJ1654 to LJ1656, encoding a sugar phosphotransferase system (PTS) transporter annotated as mannose PTS; and LJ1680, whose product shares 30% amino acid identity with immunoglobulin A proteases from pathogenic bacteria. Knockout mutants were tested in vivo. The experiments revealed that deletion of LJ1654 to LJ1656 and LJ1680 decreased the gut residence time, while a mutant with a deleted exopolysaccharide biosynthesis cluster had a slightly increased residence time.

* Corresponding author. Mailing address: Nestlé Research Centre, Nutrition and Health Department/Food and Health Microbiology, CH-1000 Lausanne 26 Vers-chez-les-Blanc, Switzerland. Phone: 0041 21 785 8676. Fax: 0041 21 785 8544. E-mail: harald.bruessow{at}rdls.nestle.com

{triangledown} Published ahead of print on 25 January 2008.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, May 2008, p. 3161-3168, Vol. 190, No. 9
0021-9193/08/$08.00+0     doi:10.1128/JB.01637-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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