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Journal of Bacteriology, August 1998, p. 4154-4159, Vol. 180, No. 16
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Structural and Functional Analysis of the Gene Cluster Encoding the Enzymes of the Arginine Deiminase Pathway of Lactobacillus sake

Manuel Zúñiga,1,dagger Marie Champomier-Verges,2 Monique Zagorec,2 and Gaspar Pérez-Martínez1,*

Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de Alimentos (CSIC), 46100 Burjassot, Valencia, Spain,1 and Laboratoire de Recherches sur la Viande, INRA-Jouy, Domaine de Vilvert, 78352 Jouy en Josas, France2

Received 29 December 1997/Accepted 3 June 1998

Lactobacillus sake can use arginine via the arginine deiminase (ADI) pathway. We designed degenerate primers based on an alignment of known sequences of ornithine transcarbamoylase (OTC)-encoding genes in order to amplify the L. sake counterpart sequences by PCR. Screening a genomic library of L. sake in lambda EMBL3 allowed us to isolate a clone containing a 10-kb L. sake genomic DNA insert. Sequence analysis revealed that the genes involved in arginine catabolism were clustered and encoded ADI (arcA), OTC (arcB), carbamate kinase (arcC), and a putative carrier with high similarity to the arginine/ornithine antiporter of Pseudomonas aeruginosa (arcD). Additionally, a putative transaminase-encoding gene (arcT) was located in this region. The genes followed the order arcA arcB arcC arcT arcD, which differs from that found in other microorganisms. arcA, arcB, arcC, and arcD mutants were constructed, and the ADI pathway was impaired in all of them. Transcriptional studies indicated that arcA gene is subject to catabolite repression, and under the conditions used, several transcripts could be detected, suggesting the existence of different initiation sites or processing of a larger mRNA.

* Corresponding author. Mailing address: Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Polígono de la Coma s/n, Apartado de Correos 73, 46100 Burjassot, Valencia, Spain. Phone: 34 6 390 00 22. Fax: 34 6 3636301. E-mail: gaspar.perez{at}iata.csic.es.

dagger Present address: Department of Molecular Genetics, Biologisch Centrum, 9750 AA Haren, The Netherlands.

Journal of Bacteriology, August 1998, p. 4154-4159, Vol. 180, No. 16
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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