title: Genetic and biochemical analysis of a class C non-specific acid phosphatase (NSAP) of Clostridium perfringens.
abstract: Clostridium perfringens, a Gram-positive anaerobe, is a human pathogen that causes gas gangrene in muscle tissues. Its ability to grow and survive in the host is believed to be due to the production of numerous enzymes that enable the organism to obtain essential nutrients from the host. In this study, CPE0201, a putative acid phosphatase gene deduced by genome analysis, was shown to encode a non-specific acid phosphatase in C. perfringens. Multiple alignments of the amino acid sequence showed that CPE0201 shares two signature motifs that belong to a class C acid phosphatase family. Expression of CPE0201 was shown to be positively regulated by the global VirR/VirS-VR-RNA regulatory cascade at the transcriptional level. To determine the acid phosphatase activity of the CPE0201-encoded protein, cloning, expression, purification and several biochemical characterizations were carried out. The optimum pH for activity of the CPE0201 enzyme was 4.8, and its V(max) and K(m) were 3.08 nmol ml(-1) min(-1) and 2.84 mM, respectively, with p-nitrophenyl phosphate (PNPP) as substrate. A CPE0201 mutant did not grow in a minimal medium containing PNPP, while it showed normal growth when Na(2)HPO(4) was added to the medium. The enzyme appears to be associated with the surface of the cell, where it may function to acquire inorganic phosphate from organic phosphomonoesters in acidic conditions, which could play an important role in the survival and growth of C. perfringens in the host tissue.
authors.affiliation: Department of Bacteriology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8640, Japan.
authors.forename: Ruoyu
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Bacteriology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8640, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Ohtani
authors.affiliation: Department of Bacteriology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8640, Japan.
authors.forename: Yun
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Bacteriology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8640, Japan.
authors.forename: Yonghui
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: Department of Bacteriology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8640, Japan.
authors.forename: Sufi
authors.initials: S
authors.lastname: Hassan
authors.affiliation: Department of Bacteriology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8640, Japan.
authors.forename: Tohru
authors.initials: T
authors.lastname: Shimizu
mesh_terms: D000135:Acid Phosphatase / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003016:Clostridium perfringens / Q000201:enzymology / Q000235:genetics*; D003470:Culture Media; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009596:Nitrophenols / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D009596:Nitrophenols; D009943:Organophosphorus Compounds; C008644:nitrophenylphosphate; D000135:Acid Phosphatase