Abstract The poultry industry is a progressive and prospective agro-based sector in Bangladesh. Poultry droppings (PD) make an excellent and abundant raw material for anaerobic co-digestion (AD) because of its high nitrogen content. Two sets of comparative assays were conducted on the anaerobic co-digestion of PD with two lignocellulosic co-substrates (LCSs), namely wheat straw (WS) and meadow grass (MG), under five different mixing ratios to optimize substrate composition and C:N ratio for enhanced biogas production. All digesters were run simultaneously under a mesophilic temperature of 35 ± 1 °C with an identical volatile solids (VS) concentration. The results showed that the co-digestion of PD with LCSs was significantly higher in terms of biogas yield and bio-methane potential (BMP) than those obtained by mono-digestion of PD and LCSs. Co-digestion of PD and MG produced a higher cumulative biogas production, biogas yield and BMP than from respectively PD and WS. The highest methane contents found were 330.1 and 340.1 Nl kg − 1 VS after digestion for 90 days at a mixing ratio of, respectively, 70:30 (PD:WS) with a C:N ratio of 32.02 and a mixing ratio of 50:50 (PD:MG) with a C:N ratio of 31.52. The increases were 1.14 and 1.13 times those of the LCSs alone, respectively. Predicted optimum ratio for PD:LCSs and C:N ratios, maximum BMP and percentage volatile solids destruction (PVSD) were calculated by using software MINITAB-17 according to the best fit regression models for co-digestion of PD with LCSs.