The proteomics analysis could contribute to better understand about metabolic pathways in anaerobic digestion community because it still as a “black-box” process. This study aimed to analyze the proteins of the anaerobic co-digestion performed in reactors containing residues from the first and second generation ethanol production. Metaproteomics analysis was carried out for three types of samples: anaerobic sludge without substrate (SI), semi-continuous stirred reactor (s-CSTR) with co-digestion of filter cake, vinasse, and deacetylation liquor (R-CoAD) and s-CSTR with co-digestion of these aforementioned residues adding Fe3O4 nanoparticles (R-NP). The R-CoAD reactor achieved 234 NmLCH4 gVS−1 and 65% of CH4 in the biogas, while the R-NP reactor reached 2800 NmLCH4 gVS−1 and 80% of CH4. The main proteins found were enolase, xylose isomerase, pyruvate phosphate dikinase, with different proportion in each sample, indicating some change in pathways. However, according to those identified proteins, the main metabolic routes involved in the co-digestion was the syntrophic acetate oxidation coupled with hydrogenotrophic methanogenesis, with the CH4 production occurring preferentially via CO2 reduction. These findings contributed to unravel the anaerobic co-digestion at a micromolecular level, and may select a more appropriate inoculum for biogas production according to that residue, reducing reaction time and increasing productivity.