The terminal oxidoreductase of
nitrous
oxide respiration in the marine, denitrifying bacterium, Pseudomonas
perfectomarinus, was identified as multi-copper protein and
purified
to electrophoretic homogeneity. The enzyme reduced N20
to N2 with hydrogen, clostridial hydrogenase,
and
methyl viologen as electron-donating system. The copper content of the
reductase corresponded to ~8 copper atoms/120000
Mr.
The subunit structure was dimeric with two peptides of equal size.
Manganese,
iron and zinc were absent, or were not found in stoichiometric amounts.
The oxidized chromophore had absorption maxima at 350, 480, 530, 620
and
780 nm; addition of dithionite produced a blue protein form with maxima
at 470, 635 and 740 nm. Both forms of the enzyme were paramagnetic. The
same copper protein was also isolated from Pseudomonas stutzeri.