title: The molecular class C acid phosphatase of Chryseobacterium meningosepticum (OlpA) is a broad-spectrum nucleotidase with preferential activity on 5'-nucleotides.
abstract: The olpA gene of Chryseobacterium meningosepticum, encoding a molecular class C phosphatase, was cloned and expressed in Escherichia coli. The gene encodes a 29-kDa polypeptide containing an amino-terminal signal peptide typical of bacterial membrane lipoproteins. Expression in E. coli results in a functional product that mostly partitions in the outer membrane. A secreted soluble OlpA derivative (sOlpA) lacking the N-terminal cysteine residue for lipid anchoring was produced in E. coli and purified by means of two steps of ion exchange chromatography. Analysis of the kinetic parameters of sOlpA with several organic phosphoesters revealed that the enzyme was able to efficiently hydrolyze nucleotide monophosphates, with a strong preference for 5'-nucleotides and for 3'-AMP. The enzyme was also able to hydrolyze sugar phosphates and beta-glycerol phosphate, although with a lower efficiency, whereas it was apparently inactive against nucleotide di- and triphosphates, diesters, and phytate. OlpA, therefore, can be considered a broad-spectrum nucleotidase with preference for 5'-nucleotides. Its functional behaviour exhibits differences from that of the Haemophilus influenzae OMP P4 lipoprotein, revealing functional heterogeneity among phosphatases of molecular class C.
authors.affiliation: Dipartimento di Scienze di Sanità Pubblica, Sezione di Microbiologia, Università di Roma La Sapienza, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Passariello
authors.forename: Serena
authors.initials: S
authors.lastname: Schippa
authors.forename: Patrizia
authors.initials: P
authors.lastname: Iori
authors.forename: Francesca
authors.initials: F
authors.lastname: Berlutti
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Thaller
authors.forename: Gian Maria
authors.initials: GM
authors.lastname: Rossolini
mesh_terms: D000135:Acid Phosphatase / Q000302:isolation & purification / Q000378:metabolism*; D001419:Bacteria / Q000201:enzymology*; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D006863:Hydrogen-Ion Concentration; D013379:Substrate Specificity / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D000135:Acid Phosphatase