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Applied and Environmental Microbiology, February 2002, p. 720-727, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.720-727.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification, Characterization, and Expression of a Second, Bicistronic, Operon Involved in the Production of Lactocin S in Lactobacillus sakei L45

Morten Skaugen,* Elizabeth L. Andersen, Vigdis H. Christie, and Ingolf F. Nes

Laboratory of Microbial Gene Technology, Agricultural University of Norway, Ås, Norway

Received 2 August 2001/ Accepted 14 November 2001

Through the analysis of spontaneous insertion mutants of Lactobacillus sakei L45, a second operon involved in lactocin S production was identified and characterized. The new, bicistronic unit, termed lasXY, is situated immediately upstream of the previously characterized nine-open reading frame (ORF) lactocin S operon (lasA-W) and is transcribed in the opposite direction. The proximal of the two newly identified genes, lasX, specifies a 285-residue protein that is similar to a group of proteins with reported gene regulation functions in gram-positive bacteria. It was demonstrated that the lasX mutants have a strongly reduced level of lasA and lasA-W mRNA, thus indicating the likely cause of the Bac- phenotype of these mutants. The second ORF in the operon, lasY, specifies a 300-residue ABC transporter homolog, the function of which is currently obscure. Transcription initiation mapping of the lasXY operon demonstrates that the two lactocin S promoters overlap such that both transcripts initiate within the -35 region of the oppositely oriented promoter. This organization of promoters is unique among this group of regulons and may constitute a modulatory site in the proposed LasX-dependent expression of lasA and downstream genes.

* Corresponding author. Mailing address: Laboratory of Microbial Gene Technology, Agricultural University of Norway, P.O. Box 5051, N-1432 Ås-NLH, Norway. Phone: 47-64949466. Fax: 47-64941465. E-mail: morten.skaugen{at}ikb.nlh.no.

Applied and Environmental Microbiology, February 2002, p. 720-727, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.720-727.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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