title: Proteus mirabilis amino acid deaminase: cloning, nucleotide sequence, and characterization of aad.
abstract: Proteus, Providencia, and Morganella species produce deaminases that generate alpha-keto acids from amino acids. The alpha-keto acid products are detected by the formation of colored iron complexes, raising the possibility that the enzyme functions to secure iron for these species, which do not produce traditional siderophores. A gene encoding an amino acid deaminase of uropathogenic Proteus mirabilis was identified by screening a genomic library hosted in Escherichia coli DH5 alpha for amino acid deaminase activity. The deaminase gene, localized on a cosmid clone by subcloning and Tn5::751 mutagenesis, was subjected to nucleotide sequencing. A single open reading frame, designated aad (amino acid deaminase), which appears to be both necessary and sufficient for deaminase activity, predicts a 473-amino-acid polypeptide (51,151 Da) encoded within an area mapped by transposon mutagenesis. The predicted amino acid sequence of Aad did not share significant amino acid sequence similarity with any other polypeptide in the PIR or SwissProt database. Amino acid deaminase activity in both P. mirabilis and E. coli transformed with aad-encoding plasmids was not affected by medium iron concentration or expression of genes in multicopy in fur, cya, or crp E. coli backgrounds. Enzyme expression was negatively affected by growth with glucose or glycerol as the sole carbon source but was not consistent with catabolite repression.
authors.affiliation: Division of Infectious Diseases, University of Maryland School of Medicine, Baltimore 21201, USA.
authors.forename: G
authors.initials: G
authors.lastname: Massad
authors.forename: H
authors.initials: H
authors.lastname: Zhao
authors.forename: H L
authors.initials: HL
authors.lastname: Mobley
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D003001:Cloning, Molecular; D003641:Deamination; D005290:Ferric Compounds / Q000378:metabolism; D018628:Gene Dosage; D045782:Hemiterpenes; D006801:Humans; D007651:Keto Acids / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011512:Proteus Infections / Q000382:microbiology; D011513:Proteus mirabilis / Q000201:enzymology / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D017262:Siderophores / Q000032:analysis; D013045:Species Specificity
chemical_list: D005290:Ferric Compounds; D045782:Hemiterpenes; D007651:Keto Acids; D011994:Recombinant Proteins; D017262:Siderophores; C001505:alpha-ketoisovalerate; C013082:alpha-ketoisocaproic acid; D000594:Amino Acid Oxidoreductases