Characterization of a unique class C acid phosphatase from Clostridium perfringens.
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 approximately 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 K(m)s ranged from 0.2 to 0.6 mM with maximum velocity ranging from 0.8 to 1.6 micromol of P(i)/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.
Reilly
Thomas J
TJ
Department of Veterinary Pathobiology, University of Missouri, Columbia, 65211, USA. reillyt@missouri.edu
Chance
Deborah L
DL
Calcutt
Michael J
MJ
Tanner
John J
JJ
Felts
Richard L
RL
Waller
Stephen C
SC
Henzl
Michael T
MT
Mawhinney
Thomas P
TP
Ganjam
Irene K
IK
Fales
William H
WH
D000135:Acid Phosphatase / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002413:Cations, Divalent / Q000494:pharmacology; D003016:Clostridium perfringens / Q000201:enzymology*; D019281:Dimerization; D020536:Enzyme Activators / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006923:Hymecromone / Q000031:analogs & derivatives; D007700:Kinetics; D008970:Molecular Weight; D009596:Nitrophenols / Q000378:metabolism; D009705:Nucleosides; D009943:Organophosphorus Compounds / Q000378:metabolism; D013379:Substrate Specificity
D001426:Bacterial Proteins; D002413:Cations, Divalent; D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D009596:Nitrophenols; D009705:Nucleosides; D009943:Organophosphorus Compounds; C008644:nitrophenylphosphate; C005359:4-methylumbelliferyl phosphate; D006923:Hymecromone; D000135:Acid Phosphatase