Despite the development of biorefinery processes, so far this area does not take into account the possibility of coupling the “conventional” composting process with the production of biochemicals. However, net carbon monoxide (CO) production has been observed during bio-waste composting. So far, O2 concentration and temperature have been identified as main variables influencing CO formation. This study aimed to investigate the CO net production during bio-waste composting under controlled laboratory conditions, depending on different aeration rates and temperatures. A series of composting processes were carried out in conditions ranging from ~psychrophilic to thermophilic (T=35, 45, 55, and 65°C) and an aeration rate of 2.7, 3.4, 4.8, and 7.8 L‧h-1. The highest concentrations of CO in each thermal variant was achieved with oxygen deficit (aeration rate 2.7 L‧h-1); additionally, CO levels increased with temperature, reaching ~300 ppm at 65°C. Production of CO in mesophilic and thermophilic conditions draws attention to the biological CO formation by microorganisms capable of producing the CODH enzyme. Further research on the CO production efficiency in these thermal ranges is necessary with the characterization of the microbial community and analysis of the ability of the identified bacteria to produce the CODH enzyme and convert CO from CO2.