Structural and functional analysis of the gene cluster encoding the enzymes of the arginine deiminase pathway of Lactobacillus sake

• Published in: Journal of bacteriology Vol. 180; no. 16; pp. 4154 - 4159
• Main Authors: Zuniga, M. (CSIC, Valencia, Spain.), Champomier-Verges, M, Zagorec, M, Perez-Martinez, G
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 01.08.1998
• Subjects: ACIDE AMINE; amino acid metabolism; AMINO ACID SEQUENCES; AMINO ACIDS; AMINOACIDOS; AMINOTRANSFERASAS; AMINOTRANSFERASE; AMINOTRANSFERASES; ARGININA; ARGININE; ARN MENSAJERO; ARN MESSAGER; BACTERIAL PROTEINS; Bacterial Proteins - genetics; Bacteriology; Base Sequence; CHEMICAL COMPOSITION; CLONACION; CLONAGE; CLONING; Cloning, Molecular; COMPOSICION QUIMICA; COMPOSITION CHIMIQUE; DNA, Bacterial; enzymology; Escherichia coli Proteins; EXPRESION GENICA; EXPRESSION DES GENES; GENBANK/AJ001330; GENE; Gene Amplification; GENE EXPRESSION; GENES; Genes, Bacterial; GENETICA; GENETICS; Genetics and Molecular Biology; GENETIQUE; Genomic Library; HIDROLASAS; HYDROLASE; HYDROLASES; Hydrolases - genetics; KINASES; LACTOBACILLUS; Lactobacillus - enzymology; Lactobacillus - genetics; Membrane Proteins; Membrane Proteins - genetics; MESSENGER RNA; METABOLISME DES PROTEINES; METABOLISMO PROTEICO; MICROBIAL PROTEINS; MOLECULAR SEQUENCE DATA; Multigene Family; Mutagenesis; NUCLEOTIDE SEQUENCE; nucleotide sequences; ORNITHINE CARBAMOYLTRANSFERASE; phosphotransferases (kinases); Protein Kinases; PROTEIN METABOLISM; PROTEINAS MICROBIANAS; PROTEINE MICROBIENNE; RESTRICTION MAPPING; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; STRUCTURAL GENES; transaminases; Transcription, Genetic; TRANSFERASAS; TRANSFERASE; TRANSFERASES
• ISSN: 0021-9193; 1098-5530
• DOI: 10.1128/jb.180.16.4154-4159.1998

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