Raw biogas can be an alternative feedstock to pure methane (CH4) for methanol production. In this investigation, we evaluated the methanol production potential of Methylosinus sporium from raw biogas originated from an anaerobic digester. Furthermore, the roles of different gases in methanol production were investigated using synthetic gas mixtures of CH4, carbon dioxide (CO2), and hydrogen (H-2). Maximum methanol production was 5.13, 4.35, 6.28, 7.16, 0.38, and 0.36 mM from raw biogas, CH4: CO2, CH4: H2, CH4: CO2: H-2, CO2, and CO2: H-2, respectively. Supplementation of H-2 into raw biogas increased methanol production up to 3.5-fold. Additionally, covalent immobilization of M. sporium on chitosan resulted in higher methanol production from raw biogas. This study provides a suitable approach to improve methanol production using low cost raw biogas as a feed containing high concentrations of H2S (0.13%). To our knowledge, this is the first report on methanol production from raw biogas, using immobilized cells of methanotrophs. (C) 2016 Elsevier Ltd. All rights reserved.