Characterization of a Unique Class C Acid Phosphatase from Clostridium perfringens

• Published in: Applied and Environmental Microbiology 2009-06, Vol.75 (11), p.3745-3754
• Main Authors: Reilly, Thomas J, Chance, Deborah L, Calcutt, Michael J, Tanner, John J, Felts, Richard L, Waller, Stephen C, Henzl, Michael T, Mawhinney, Thomas P, Ganjam, Irene K, Fales, William H
• Format: Article
• Language: English
• Subjects: Acid phosphatase; Acid Phosphatase - chemistry; Acid Phosphatase - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; Biological and medical sciences; Cations, Divalent - pharmacology; Clostridium perfringens; Clostridium perfringens - enzymology; Dimerization; Enzyme Activators - pharmacology; Enzyme Inhibitors - pharmacology; Enzymes; Enzymology and Protein Engineering; Fundamental and applied biological sciences. Psychology; Gram-positive bacteria; Hymecromone - analogs & derivatives; Kinetics; Lipids; Microbiology; Molecular Weight; Nitrophenols - metabolism; Nucleosides; Organophosphorus Compounds - metabolism; Phosphates; Substrate Specificity
• ISSN: 0099-2240; 1098-5336; 1098-6596
• DOI: 10.1128/AEM.01599-08

Clostridium perfringens is a gram-positive anaerobe and a pathogen of medical importance. The detection of acid phosphatase activity is a powerful diagnostic indicator of the presence of C. perfringens among anaerobic isolates; however, characterization of the enzyme has not previously been reported. Provided here are details of the characterization of a soluble recombinant form of this cell-associated enzyme. The denatured enzyme was ~31 kDa and a homodimer in solution. It catalyzed the hydrolysis of several substrates, including para-nitrophenyl phosphate, 4-methylumbelliferyl phosphate, and 3' and 5' nucleoside monophosphates at pH 6. Calculated Kms ranged from 0.2 to 0.6 mM with maximum velocity ranging from 0.8 to 1.6 μmol of Pi/s/mg. Activity was enhanced in the presence of some divalent cations but diminished in the presence of others. Wild-type enzyme was detected in all clinical C. perfringens isolates tested and found to be cell associated. The described enzyme belongs to nonspecific acid phosphatase class C but is devoid of lipid modification commonly attributed to this class.