Staphylococcus haemolyticus contains two D-glutamic acid biosynthetic activities, a glutamate racemase and a D-amino acid transaminase.
Two D-glutamic acid biosynthetic activities, glutamate racemase and D-amino acid transaminase, have been described previously for bacteria. To date, no bacterial species has been reported to possess both activities. Genetic complementation studies using Escherichia coli WM335, a D-glutamic acid auxotroph, and cloned chromosomal DNA fragments from Staphylococcus haemolyticus revealed two distinct DNA fragments containing open reading frames which, when present, allowed growth on medium without exogenous D-glutamic acid. Amino acid sequences of the two open reading frames derived from the DNA nucleotide sequences indicated extensive identity with the amino acid sequence of Pediococcus pentosaceous glutamate racemase in one case and with that of the D-amino acid transaminase of Bacillus spp. in the second case. Enzymatic assays of lysates of E. coli WM335 strains containing either the cloned staphylococcal racemase or transminase verified the identities of these activities. Subsequent DNA hybridization experiments indicated that Staphylococcus aureus, in addition to S. haemolyticus, contained homologous chromosomal DNA for each of these genes. These data suggest that S. haemolyticus, and probably S. aureus, contains genes for two D-glutamic acid biosynthetic activities, a glutamate racemase (dga gene) and a D-amino acid transaminase (dat gene).
Pucci
M J
MJ
Department of Microbiology, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, Connecticut 06492-7660.
Thanassi
J A
JA
Ho
H T
HT
Falk
P J
PJ
Dougherty
T J
TJ
D000591:Amino Acid Isomerases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D051263:D-Alanine Transaminase; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D018698:Glutamic Acid / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011994:Recombinant Proteins / Q000096:biosynthesis; D013210:Staphylococcus / Q000201:enzymology* / Q000235:genetics; D000637:Transaminases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D018698:Glutamic Acid; D000637:Transaminases; D051263:D-Alanine Transaminase; D000591:Amino Acid Isomerases; C062292:glutamate racemase