In Greece biomass is often being disposed of uncontrollably, resulting in significant environmental impacts. The aim of this study is the single-stage anaerobic co-digestion assessment, valorizing Northern and Southern Greece mixtures, resulting from previous literature reviews, experimental designs, and biochemical methane potential (BMP) assays. Regarding the methane yield maximization, in Northern Greece, the most suitable mixture was 10% corn silage, 80% cattle manure, and 10% malt; while in Southern Greece it was 10% corn silage, 57% cattle manure, 23% orange peels, and 10% olive pomace for fall/winter season. The hydraulic retention time (HRT) was set at 20 d and an initial organic loading rate (OLR) of 2 g COD/(L·d) was applied, with a view to gradually increase it. However, volatile fatty acids accumulation was observed, which led to OLR reduction to 1.5 g COD/(L·d) for both experiments. The Northern Greece reactor operated successfully for OLR 1.5–5 g COD/(L·d), while further increase led to system failure. On the other hand, the reactor of the Southern Greece mixture operated successfully at OLR 1.5–2 g COD/(L·d), but further operation indicated inadequacy, probably due to inhibitor (such as limonene) accumulation. Mixtures consisting of corn silage, cattle manure, and malt can be successfully valorized at high OLR. However, further investigation for mixtures with orange peels is suggested due to the presence of inhibitors.