Cyanobacteria (blue-green algae) are oxygenic phototrophic bacteria carryin
g out plant-type photosynthesis. The only hydrogen peroxide scavenging enzy
mes in at least two unicellular species have been demonstrated to be bifunc
tional cytosolic catalase-peroxidases (CatPXs) having considerable homology
at the active site with plant ascorbate peroxidases (APXs). In this paper
we examined optical and kinetic properties of CatPXs from the cyanobacteria
Anacystis nidulans and Synechocystis PCC 6803 and discuss similarities and
differences to plant APXs. Both CatPXs and APX showed similar spectra of t
he ferric enzyme, the redox intermediate Compound I and the cyanide complex
, whereas the spectrum of CatPX Compound II had characteristics reminescent
of the spectrum of the native enzyme. Both steady-state and multi-mixing t
ransient-state kinetic studies were performed in order to characterize the
kinetic behaviour of CatPXs. Bimolecular rate constants of both formation a
nd reduction of a CatPX Compound I are presented. Because of its intrinsic
high catalase activity (which cannot be found in APXs), the rate constants
for Compound I formation were measured with peroxoacetic acid and are shown
to be 5.9 x 10(4) M-1 s(-1) for CatPX from A. nidulans and 8.7 x 10(3) M-1
s(-1) for the Synechocystis enzyme. Compared with o-dianisidine (2.7-6.7 x
10(6) M(-1)s(-1)) and pyrogallol (8.6 x 10(4)-1.6 x 10(5) M-1 s(-1)), the
rate constant for Compound I reduction by ascorbate was extremely low (5.4
x 10(3) M-1 s(-1) at pH 7.0 and 15 degrees C), in marked contrast to the be
haviour of APXs.