Bromoperoxidase activity of vanadate-substituted acid phosphatases from Shigella flexneri and Salmonella enterica ser. typhimurium.
Vanadium haloperoxidases and the bacterial class A nonspecific acid phosphatases have a conserved active site. It is shown that vanadate-substituted recombinant acid phosphatase from Shigella flexneri (PhoN-Sf) and Salmonella enterica ser. typhimurium (PhoN-Se) in the presence of H2O2 are able to oxidize bromide to hypobromous acid. Vanadate is essential for this activity. The kinetic parameters for the artificial bromoperoxidases have been determined. The Km value for H2O2 is about the same as that for the vanadium bromoperoxidases from the seaweed Ascophyllum nodosum. However, the Km value for Br- is about 10-20 times higher, and the turnover values of about 3.4 min-1 and 33 min-1 for PhoN-Sf and PhoN-Se, respectively, are much slower, than those of the native bromoperoxidase. Thus, despite the striking similarity in the active-site structures of the vanadium haloperoxidases and the acid phosphatase, the turnover frequency is low, and clearly the active site of acid phosphatases is not optimized for haloperoxidase activity. Like the native vanadium bromoperoxidase, the vanadate-substituted PhoN-Sf and PhoN-Se catalyse the enantioselective sulfoxidation of thioanisole.
Tanaka
Naoko
N
Institute for Molecular Chemistry, University of Amsterdam, The Netherlands.
Dumay
Valérie
V
Liao
Qianning
Q
Lange
Alex J
AJ
Wever
Ron
R
D000135:Acid Phosphatase / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D010544:Peroxidases / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012486:Salmonella typhimurium / Q000201:enzymology*; D012763:Shigella flexneri / Q000201:enzymology*; D013379:Substrate Specificity / Q000235:genetics; D014638:Vanadates / Q000378:metabolism
D011994:Recombinant Proteins; D014638:Vanadates; D010544:Peroxidases; C026195:bromide peroxidase; D000135:Acid Phosphatase