Abstract
The release of glucose from lignocellulosic waste for subsequent fermentation into biofuels holds promise for securing humankind's future energy needs. The discovery of a set of copper-dependent enzymes known as lytic polysaccharide monooxygenases (LPMOs) has galvanised new research in this area. LPMOs act by oxidatively introducing chain breaks into cellulose and other polysaccharides, boosting the ability of cellulases to act on the substrate. Although several proteins have been implicated as electron sources in fungal LPMO biochemistry, no equivalent bacterial LPMO electron donors have been previously identified, although the proteins Cbp2D and E from
Cellvibrio japonicus
have been implicated as potential candidates. Here we analyse a small
c
-type cytochrome (
Cj
X183) present in
Cellvibrio japonicus
Cbp2D, and show that it can initiate bacterial Cu
II/I
LPMO reduction and also activate LPMO-catalyzed cellulose-degradation. In the absence of cellulose,
Cj
X183-driven reduction of the LPMO results in less H
2
O
2
production from O
2
, and correspondingly less oxidative damage to the enzyme than when ascorbate is used as the reducing agent. Significantly, using
Cj
X183 as the activator maintained similar cellulase boosting levels relative to the use of an equivalent amount of ascorbate. Our results therefore add further evidence to the impact that the choice of electron source can have on LPMO action. Furthermore, the study of Cbp2D and other similar proteins may yet reveal new insight into the redox processes governing polysaccharide degradation in bacteria.