Properties of hexahistidine-tagged organophosphate hydrolase.
The catalytic properties of organophosphate hydrolase (OPH) containing a hexahistidine tag His6 (His6-OPH) and purified to 98% homogeneity were investigated. The pH optimum of enzymatic activity and isoelectric point of His6-OPH, which were shown to be 10.5 and 8.5, respectively, are shifted to the alkaline range as compared to the same parameters of the native OPH. The recombinant enzyme possessed improved catalytic activity towards S-containing substrates: the catalytic efficiency of methylparathion hydrolysis by His6-OPH is 4.2 x 10(6) M(-1) x sec(-1), whereas by native OPH it is 3.5 x 10(5) M(-1) x sec(-1).
Votchitseva
Yu A
YA
Faculty of Chemistry, Lomonosov Moscow State University, 119992 Moscow, Russia.
Efremenko
E N
EN
Aliev
T K
TK
Varfolomeyev
S D
SD
D020134:Catalytic Domain; D003035:Cobalt / Q000737:chemistry; D004586:Electrophoresis; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006639:Histidine / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity; D013696:Temperature
C471213:His-His-His-His-His-His; D009842:Oligopeptides; D003035:Cobalt; D006639:Histidine; C045290:phosphorylphosphatase; D010744:Phosphoric Monoester Hydrolases