Document Detail


Genetic and biochemical analysis of a class C non-specific acid phosphatase (NSAP) of Clostridium perfringens.
MedLine Citation:
PMID:  19833778     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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:
Ruoyu Wang; Kaori Ohtani; Yun Wang; Yonghui Yuan; Sufi Hassan; Tohru Shimizu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-10-15
Journal Detail:
Title:  Microbiology (Reading, England)     Volume:  156     ISSN:  1465-2080     ISO Abbreviation:  Microbiology (Reading, Engl.)     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2009-12-23     Completed Date:  2010-02-25     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9430468     Medline TA:  Microbiology     Country:  England    
Other Details:
Languages:  eng     Pagination:  167-73     Citation Subset:  IM    
Affiliation:
Department of Bacteriology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8640, Japan.
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MeSH Terms
Descriptor/Qualifier:
Acid Phosphatase / genetics,  metabolism*
Amino Acid Sequence
Bacterial Proteins / genetics,  metabolism*
Cloning, Molecular
Clostridium perfringens / enzymology,  genetics*
Culture Media
Gene Expression Regulation, Bacterial
Molecular Sequence Data
Nitrophenols / metabolism
Organophosphorus Compounds / metabolism
Phylogeny
Sequence Alignment
Sequence Analysis, DNA
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/Culture Media; 0/Nitrophenols; 0/Organophosphorus Compounds; 330-13-2/nitrophenylphosphate; EC 3.1.3.2/Acid Phosphatase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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