Purification and characterization of a sialidase from Clostridium chauvoei NC08596.
The sialidase secreted by Clostridium chauvoei NC08596 was purified to apparent homogeneity by ion-exchange chromatography, gel filtration, hydrophobic interaction-chromatography, FPLC ion-exchange chromatography, and FPLC gel filtration. The enzyme was enriched about 10,200-fold, reaching a final specific activity of 24.4 U mg-1. It has a relatively high molecular mass of 300 kDa and consists of two subunits each of 150 kDa. The cations Mn2+, Mg2+, and Ca2+ and bovine serum albumin have a positive effect on the sialidase activity, while Hg2+, Cu2+, and Zn2+, chelating agents and salt decrease enzyme activity. The substrate specificity, kinetic data, and pH optimum of the enzyme are similar to those of other bacterial sialidases.
Heuermann
D
D
Biochemisches Institut, Christian-Albrechts-Universität, Kiel, Germany.
Roggentin
P
P
Kleineidam
R G
RG
Schauer
R
R
D003013:Clostridium / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009439:Neuraminidase / Q000737:chemistry / Q000302:isolation & purification*; D013379:Substrate Specificity
D009439:Neuraminidase