This study assessed the co-digestion of macro and microalgal biomass towards the improvement of hydrogen production. The red macroalgal biomass (Gelidium amansii) and green mixed microalgal biomass was mixed in a ratio of 8:2, with an initial substrate concentration of 10 g/L, and various amount of inoculum addition range from 3 to 15% (v/v) was evaluated to assess the feasible substrate to inoculum ratio for the effective co-digestion of the algal biomass. The results showed that the co-digestion with 6% inoculum addition provided the peak hydrogen yield of 45 mL/g dry biomass added with a high hydrogen content of 24% in the gas phase. The other tested conditions showed moderate hydrogen content in the range of 17–22%, respectively. These results suggest that anaerobic co-digestion of macro and microalgal biomass, with appropriate initial biomass loading (6%) is essential for enhanced hydrogen production.