Expression of the vanadium-dependent bromoperoxidase gene from a marine macro-alga Corallina pilulifera in Saccharomyces cerevisiae and characterization of the recombinant enzyme.
The vanadium-dependent bromoperoxidase from the marine macro-alga Corallina pilulifera was heterologously expressed in Saccharomyces cerevisiae. The enzyme was purified and crystals in "tear drop" form were obtained. The catalytic properties of the recombinant enzyme were studied and compared with those of the native enzyme purified from C. pilulifera. Differences in thermal stability and chloroperoxidase activity were observed. The recombinant enzyme retained full activity after preincubation at 65 degrees C for 20 min, but the native enzyme was completely inactivated under the same conditions. The chlorinating activity of the native enzyme was more than ten times higher than that of the recombinant enzyme. Other properties, such as K(m) values for KBr and H(2)O(2), and optimal temperature and pH, were similar for each source of C. pilulifera bromoperoxidase.
Ohshiro
Takashi
T
Department of Biotechnology, Tottori University, Tottori 680-8552, Japan.
Hemrika
Wieger
W
Aibara
Toshiaki
T
Wever
Ron
R
Izumi
Yoshikazu
Y
D001965:Bromides / Q000378:metabolism; D002711:Chloride Peroxidase / Q000378:metabolism; D003460:Crystallization; D004795:Enzyme Stability; D006358:Hot Temperature; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D010544:Peroxidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017680:Potassium Compounds / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000461:Rhodophyta / Q000201:enzymology*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014639:Vanadium / Q000378:metabolism*
D001965:Bromides; D017680:Potassium Compounds; D011994:Recombinant Proteins; D014639:Vanadium; D006861:Hydrogen Peroxide; D010544:Peroxidases; C026195:bromide peroxidase; D002711:Chloride Peroxidase; C039004:potassium bromide