title: Purification and some properties of phospho-beta-galactosidase from the Gram-negative oral bacterium Leptotrichia buccalis ATCC 14201.
abstract: Phospho-beta-galactosidase (P-beta-gal; EC 3.2.1.85) is induced during growth of Leptotrichia buccalis ATCC 14201 on lactose and lactulose. The enzyme has been purified to electrophoretic homogeneity (M(r) approximately 53 kDa, pI approximately 4.8), and kinetic parameters have been determined using the chromogenic analog o-nitrophenyl-beta-D-galactopyranoside-6-phosphate as substrate. Both ATP and galactose-6-phosphate are inhibitors of P-beta-gal activity. Microsequence analysis has identified the first 32 residues from the N-terminus of the protein, and by comparative sequence alignment the enzyme can be assigned to Family 1 of the glycosylhydrolase superfamily. Polyclonal antibody against the enzyme permits the highly specific immuno-detection of P-beta-gal in cell-free extracts of L. buccalis. Although described previously in several Gram-positive species, this is the first reported purification of P-beta-gal from a Gram-negative organism.
mesh_terms: D000595:Amino Acid Sequence; D006026:Glycoside Hydrolases; D006090:Gram-Negative Bacteria / Q000201:enzymology* / Q000254:growth & development; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009055:Mouth / Q000382:microbiology*; D001616:beta-Galactosidase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D006026:Glycoside Hydrolases; D001616:beta-Galactosidase; C022361:6-phospho-beta-galactosidase