AbstractThis study aimed to enhance the production of hydrogen (H2) from anaerobic digestion of banana peels (BP) using the synergistic interaction between the ruminant bacterial strains. Seven ruminant bacterial consortia were designed from sixteen hydrogen producing ruminant bacterial strains. MOST2 (Pediococcus acidilactici, Streptococcus lutetiensis and Bacillus cereus) was the most highly producer giving 398.00 ± 21.67 ml/L on 25% banana peels. The used optimization strategy in this study resulted in maximum H2 (HMax) of 1446.67 ± 60.64 ml/L with a significant improvement of 363.48%. The maximum hydrogen production rate (Rmax) and lag time (λ) were 47.52 ml/L/h, 2.91 h by MLM, respectively, and 35.99 ml/L/h and 5.05 h by MGM, respectively. GC–MS analysis of the fermentation effluent indicated the presence of butyric acid (3.58%), acetic acid (1.81%), and agaric acid (1.70%). The study reveals the feasibility of utilizing banana peel waste as a sustainable feedstock for hydrogen production by the ruminant bacterial consortia.