The flavin cofactor within cellobiose dehydrogenase (CDH) was found to be responsible for the reduction of all electron acceptors tested. This includes cytochrome c, the reduction of which has been reported to be by the reduced heme of CDH. The heme group was shown to affect the reactivity and activation energy with respect to individual electron acceptors, but the heme group was not involved in the direct transfer of electrons to substrate. A complicated interaction was found to exist between the flavin and heme of cellobiose dehydrogenase. The addition of electron acceptors was shown to increase the rate of flavin reduction and the electron transfer rate between the flavin and heme. All electron acceptors tested appeared to be reduced by the flavin domain. The addition of ferric iron eliminated the flavin radical present in reduced CDH, as detected by low temperature ESR spectroscopy, while it increased the flavin radical ESR signal in the independent flavin domain, more commonly referred to as cellobiose:quinone oxidoreductase (CBQR). Conversely, no radical was detected with either CDH or CBQR upon the addition of methyl-1,4-benzoquinone. Similar reaction rates and activation energies were determined for methyl-1,4-benzoquinone with both CDH and CBQR, whereas the rate of iron reduction by CDH was five times higher than by CBQR, and its activation energy was 38 kJ/mol lower than that of CBQR. Oxygen, which may be reduced by either one or two electrons, was found to behave like a two-electron acceptor. Superoxide production was found only upon the inclusion of iron. Additionally, information is presented indicating that the site of substrate reduction may be in the cleft between the flavin and heme domains.