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

ABSTRACT 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. sakecounterpart sequences by PCR. Screening a genomic library of L. 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 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.

[1]  J. Michiels,et al.  The arginine deiminase pathway in Rhizobium etli: DNA sequence analysis and functional study of the arcABC genes , 1997, Journal of bacteriology.

[2]  S. Ehrlich,et al.  Single-crossover integration in the Lactobacillus sake chromosome and insertional inactivation of the ptsI and lacL genes , 1997, Applied and environmental microbiology.

[3]  V. Monedero,et al.  Establishing a model to study the regulation of the lactose operon in Lactobacillus casei. , 1997, FEMS microbiology letters.

[4]  A. Ruepp,et al.  Fermentative arginine degradation in Halobacterium salinarium (formerly Halobacterium halobium): genes, gene products, and transcripts of the arcRACB gene cluster , 1996, Journal of bacteriology.

[5]  S. Ehrlich,et al.  Carbohydrate Utilization in Lactobacillus sake , 1996, Applied and environmental microbiology.

[6]  S. Ehrlich,et al.  Efficient transformation of Lactobacillus sake by electroporation. , 1996, Microbiology.

[7]  W. Hammes,et al.  Two genes encoding the β-galactosidase of Lactobacillus sake , 1995 .

[8]  G. G. Pritchard,et al.  Occurrence of arginine deiminase pathway enzymes in arginine catabolism by wine lactic Acid bacteria , 1995, Applied and environmental microbiology.

[9]  J. Leunissen,et al.  Divergence in codon usage of Lactobacillus species. , 1994, Nucleic acids research.

[10]  M. Saier,et al.  Analysis of a cis-active sequence mediating catabolite repression in gram-positive bacteria. , 1994, Research in microbiology.

[11]  M. Gasson,et al.  Rapid isolation of genes from bacterial lambda libraries by direct polymerase chain reaction screening. , 1993, FEMS microbiology letters.

[12]  D. Haas,et al.  RNA processing modulates the expression of the arcDABC operon in Pseudomonas aeruginosa. , 1992, Journal of molecular biology.

[13]  A. Driessen,et al.  arcD, the first gene of the arc operon for anaerobic arginine catabolism in Pseudomonas aeruginosa, encodes an arginine-ornithine exchanger , 1992, Journal of bacteriology.

[14]  D. Haas,et al.  Anaerobic regulation of transcription initiation in the arcDABC operon of Pseudomonas aeruginosa , 1991, Journal of bacteriology.

[15]  R. Leer,et al.  Incompatibility of Lactobacillus Vectors with Replicons Derived from Small Cryptic Lactobacillus Plasmids and Segregational Instability of the Introduced Vectors , 1991, Applied and environmental microbiology.

[16]  W. Hammes,et al.  Lactic acid bacteria in meat fermentation , 1990 .

[17]  M. Weickert,et al.  Site-directed mutagenesis of a catabolite repression operator sequence in Bacillus subtilis. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[18]  A. Driessen,et al.  Regulation of arginine-ornithine exchange and the arginine deiminase pathway in Streptococcus lactis , 1987, Journal of bacteriology.

[19]  M. Montel,et al.  Arginine catabolism in Lactobacillus sake isolated from meat , 1987, Applied and environmental microbiology.

[20]  G. Bender,et al.  Arginine deiminase system and bacterial adaptation to acid environments , 1987, Applied and environmental microbiology.

[21]  N. Glansdorff,et al.  Biosynthesis and Metabolism of Arginine in Bacteria , 1986, Microbiological reviews.

[22]  M. Mogi,et al.  Coordinate repression of arginine aminopeptidase and three enzymes of the arginine deiminase pathway in Streptococcus mitis. , 1986, Biochemistry International.

[23]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[24]  V. Stalon,et al.  Control of enzyme synthesis in the arginine deiminase pathway of Streptococcus faecalis , 1982, Journal of bacteriology.