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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 theL. sake counterpart sequences by PCR. Screening a genomic library ofL. sake in λEMBL3 allowed us to isolate a clone containing a 10-kbL. 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 ofPseudomonas aeruginosa (arcD ). Additionally, a putative transaminase-encoding gene (arcT ) was located in this region. The genes followed the orderarcA arcB arcC arcT arcD , which differs from that found in other microorganisms.arcA ,arcB ,arcC , andarcD mutants were constructed, and the ADI pathway was impaired in all of them. Transcriptional studies indicated thatarcA 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.

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