J. Armengaud et al., A second [2Fe-2S] ferredoxin from Sphingomonas sp strain RW1 can function as an electron donor for the dioxin dioxygenase, J BACT, 182(8), 2000, pp. 2238-2244
The first step in the degradation of dibenzofuran and dibenzo-p-dioxin by S
phingomonas sp. strain RW1 is carried out by dioxin dioxygenase (DxnA1A2),
a ring-dihydroxylating enzyme. An open reading frame (fdx3) that could pote
ntially specify a new ferredoxin has been identified downstream of dxmA1A2,
a two-cistron gene (J. Armengaud, B. Happe, and K. N. Timmis, J. Bacteriol
. 180:3954-3966, 1998). In the present study, we report a biochemical analy
sis of Fdx3 produced in Escherichia coli. This third ferredoxin thus far id
entified in Sphingomonas sp. strain RW1 contained a putidaredoxin-type [2Fe
-2S] cluster which was characterized by UV-visible absorption spectrophotom
etry and electron paramagnetic resonance spectroscopy. The midpoint redox p
otential of this ferredoxin (E'(o) -247 +/- 10 mV versus normal hydrogen el
ectrode at pH 8.0) is similar to that exhibited by Fdx1 (-245 mV), a homolo
gous ferredoxin previously characterized in Sphingomonas sp. strain RW1. In
in vitro assays, Fdx3 can be reduced by RedA2 (a reductase similar to clas
s I cytochrome P-450 reductases), previously isolated from Sphingomonas sp.
strain RW1. RedA2 exhibits a K-m value of 3.2 +/- 0.3 mu M for Fdx3. In vi
vo coexpression of fdx3 and redA2 with dxnA1A2 confirmed that Fdx3 can serv
e as an electron donor for the dioxin dioxygenase.