Anp. Hiner et al., Catalase-like oxygen production by horseradish peroxidase must predominantly be an enzyme-catalyzed reaction, ARCH BIOCH, 392(2), 2001, pp. 295-302
When hydrogen peroxide (H2O2) was provided as the only substrate for horser
adish peroxidase C (HRP-C) the catalase-like emission of oxygen gas was obs
erved. The reaction was favored at neutral compared to acidic pH. Addition
of the superoxide radical scavengers tetranitromethane (TNM) or superoxide
dismutase (SOD) increased activity. TNM's effect was concentration dependen
t but SOD's was not, indicating that only some of the superoxide generated
was released into solution. Manganous ions (Mn2+) react with superoxide rad
icals to regenerate H2O2 but not oxygen; when added to the reaction medium
oxygen production was reduced but not abolished. The effect was essentially
concentration independent, suggesting that most oxygen was produced enzyma
tically and not by chemical disproportionation of superoxide. The catalase-
like activities of some site-directed mutants of HRP-C suggest that active
site residues histidine 42 and arginine 38 are influential in determining t
his activity. A clear correlation also existed between catalase activity an
d the enzymes' resistance to inactivation by H2O Computer simulation of a r
eaction scheme that included catalase-like activity agreed well with experi
mental data. (C) 2001 Academic Press.