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Applied and Environmental Microbiology, December 2006, p. 7518-7530, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.00877-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of a Putative Operon Involved in Fructooligosaccharide Utilization by Lactobacillus paracasei{triangledown} ,{dagger}

Yong Jun Goh,§ Chaomei Zhang, Andrew K. Benson, Vicki Schlegel, Jong-Hwa Lee, and Robert W. Hutkins*

Department of Food Science and Technology, University of Nebraska, Lincoln, Nebraska 68583-0919

Received 13 April 2006/ Accepted 25 September 2006

The growth and activity of some Lactobacillus and Bifidobacterium strains are stimulated by the presence of nondigestible fructooligosaccharides (FOS), which are selectively fermented by specific intestinal bacteria. Consumption of FOS, therefore, enriches for those bacteria that possess metabolic pathways necessary for FOS metabolism. In this study, a DNA microarray consisting of 7,680 random genomic library fragments of Lactobacillus paracasei 1195 was used to examine genes involved in the utilization of FOS in this organism. Differential expression profiles between cells grown on FOS and those grown on glucose provided a basis for identifying genes specifically induced by FOS. Several of the FOS-induced genes shared sequence identity with genes encoding ß-fructosidases and components of phosphoenolpyruvate-dependent phosphotransferase systems (PTS). These genes were organized in a putative operon, designated the fos operon, that may play an essential role in FOS utilization. The complete 7,631-bp nucleotide sequence of the putative fos operon was determined and consists of fosABCDXE genes, which encode a putative fructose/mannose PTS (FosABCDX) and a ß-fructosidase precursor (FosE). The latter contains an N-terminal signal peptide sequence and cell wall sorting signals at the C-terminal region, suggesting its localization at the cell wall. Inactivation of the fosE gene led to impaired growth on FOS and other ß-fructose-linked carbohydrates. Transcriptional analysis by reverse transcriptase PCR suggested that fosABCDXE was cotranscribed as a single mRNA during growth on FOS. Expression array analysis revealed that when glucose was added to FOS-grown cells, transcription of the FOS-induced genes was repressed, indicating that FOS metabolism is subject to catabolite regulation.

* Corresponding author. Mailing address: Department of Food Science and Technology, University of Nebraska, 338 FIC, Lincoln, NE 68583-0919. Phone: (402) 472-2820. Fax: (402) 472-1693. E-mail: rhutkins1{at}unl.edu.

{triangledown} Published ahead of print on 6 October 2006.

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

§ Present address: Department of Food Science, North Carolina State University, Raleigh, NC 27695-7624.

Present address: School of Bioresource Sciences, Andong National University, Andong, South Korea.

Applied and Environmental Microbiology, December 2006, p. 7518-7530, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.00877-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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