Either 110 μl of plasma-exposed (5 min treatment time) KPi buffer
(blue) or 0.7 mM H2O2 in KPi (red) were transferred to a vial containing 5 μl ethylbenzene and
Activities of free and immobilized rAaeUPO were measured using 2.5 mM ABTS, 1 mM H2O2
in 50 mM sodium acetate buffer as well as 1 or 10 nM enzyme, respectively. Activity was
Immobilized rAaeUPO
was diluted to 1 μM and 100 μl were treated with plasma for the indicated amounts of time.
Plasma treatment of rAaeUPO was performed either without
or in the presence of 1 mg ml-1 SodA for 5 min using 100 μl. Subsequently, samples were
Plasma
exposure was performed as described for HRP. Enzyme solution was then diluted 1:10 in 100
110 μl of buffer were treated for 10 min as described before and H2O2
concentration was measured using the Spectroquant kit. Then, rAaeUPO was added at 40 nM
Immobilization
was carried out as described in the Methods section. Activity assays were performed as
Plasma treatment was performed with 100 μl of 10 U ml-1 HRP with or
without the addition of 100 mM mannitol as •OH scavenger. Activities were determined as
SodA was added to the
standard reaction mixture (10 U ml-1, 5 mM guaiacol, 100 mM KPi) at 1 mg ml-1 prior to plasma
HRP was diluted to different concentrations and treated with plasma for the
indicated amounts of time (100 μl). Activity of HRP was determined subsequently using a final
