Lactose metabolism by Staphylococcus aureus: characterization of lacABCD, the structural genes of the tagatose 6-phosphate pathway.
The nucleotide and deduced amino acid sequences of the lacA and lacB genes of the Staphylococcus aureus lactose operon (lacABCDFEG) are presented. The primary translation products are polypeptides of 142 (Mr = 15,425) and 171 (Mr = 18,953) amino acids, respectively. The lacABCD loci were shown to encode enzymes of the tagatose 6-phosphate pathway through both in vitro studies and complementation analysis in Escherichia coli. A serum aldolase assay, modified to allow detection of the tagatose 6-phosphate pathway enzymes utilizing galactose 6-phosphate or fructose phosphate analogs as substrate, is described. Expression of both lacA and lacB was required for galactose 6-phosphate isomerase activity. LacC (34 kDa) demonstrated tagatose 6-phosphate kinase activity and was found to share significant homology with LacC from Lactococcus lactis and with both the minor 6-phosphofructokinase (PfkB) and 1-phosphofructokinase (FruK) from E. coli. Detection of tagatose 1,6-bisphosphate aldolase activity was dependent on expression of the 36-kDa protein specified by lacD. The LacD protein is highly homologous with LacD of L. lactis. Thus, the lacABCD genes comprise the tagatose 6-phosphate pathway and are cotranscribed with genes lacFEG, which specify proteins for transport and cleavage of lactose in S. aureus.
Rosey
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EL
Department of Microbiology, University of Kansas, Lawrence 66045.
Oskouian
B
B
Stewart
G C
GC
D000446:Aldehyde-Lyases / Q000235:genetics; D019747:Aldose-Ketose Isomerases; D000595:Amino Acid Sequence; D001483:Base Sequence; D002238:Carbohydrate Epimerases / Q000235:genetics; D005798:Genes, Bacterial; D006600:Hexosephosphates / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007785:Lactose / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010770:Phosphotransferases / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor); D012689:Sequence Homology, Nucleic Acid; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics*
D006600:Hexosephosphates; C026634:tagatose 6-phosphate; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C032258:tagatose kinase; C432824:tagatose 6-phosphate kinase; D000446:Aldehyde-Lyases; C023093:tagatose 1,6-diphosphate aldolase; D002238:Carbohydrate Epimerases; D019747:Aldose-Ketose Isomerases; C023084:galactose-6-phosphate isomerase; D007785:Lactose