Purification and partial characterization of alpha-mannosidase from Trypanosoma rangeli.
An alpha-mannosidase from Trypanosoma rangeli was partially purified by a protocol involving solubilization using lysis buffer followed by chromatography on diethylaminoethyl (DEAE)-cellulose and Sephadex 100 columns. The enzyme has a molecular weight of 45 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and of 90 kDa as determined by gel filtration. The purified T. rangeli alpha-mannosidase has a pH optimum ranging between 5 and 6 and a temperature optimum of 37 degrees C. It has activation energy of 8.15 x 10(2) J mol(-1) K(-1). The enzyme has a Michaelis constant (Km) of 99 microM for the 4-methylumbelliferyl-alpha-D-mannopyranoside substrate (MU-alpha-mann). It is strongly inhibited by swainsonine [inhibition constant (Ki) 0.048 microM] and is moderately inhibited by mannose and alpha-D-methylmannopyranoside. The enzyme activity is decreased in the presence of 1 mM Ca2+, Co2+, Cu2+, Fe2+, Fe3+, Hg2+, Mg2+, and Mn2+.
Nok
A J
AJ
Protozoology Department, Institute of Tropical Medicine, Nagasaki University, Japan.
Shuaibu
M N
MN
Kanbara
H
H
Yanagi
T
T
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007700:Kinetics; D008361:Mannosidases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D019216:Metals, Heavy / Q000494:pharmacology; D014345:Trypanosoma / Q000201:enzymology* / Q000254:growth & development; D014352:Trypanosomiasis / Q000469:parasitology; D043323:alpha-Mannosidase
D019216:Metals, Heavy; D008361:Mannosidases; D043323:alpha-Mannosidase