Hydrogen (H₂) is expected to become the most sustainable and promising clean alternative fuel in the future.  This study evaluated the potential of a new facultative anaerobic bacterium isolated from cow rumen to produce H₂ from sugarcane molasses. The bacterial isolate RM122 produced  451.67±12.14 and 387.67 ±19.23 mL/L H₂on 6% glucose and 6% molasses sugar, respectively. RM122 was characterized phenotypically, identified genotypically by 16S rRNA sequence analysis as Escherichia marmotae, and deposited in NCBI GenBank database with the accession number OP345936. H₂production was improved to 1670.00 ±40.41 mL/L by application of optimization experiments design and kinetic study. Sugarcane molasses was used as a fermentation substrate and the optimum sugar concentration was 4%, The recorded maximum hydrogen production (H_max) was 560.00±25.98 mL/L with R_max (maximum hydrogen production rate) of 31.43 ±2.14 mL/L/h, R_max (MGM)of 30.73 mL/L/h and λ (lag phase) of 15.19 h for 52.43 fermentation time. H_max of 646.67 ±23.33 mL/L with R_max(Exp) of 35.28 ±2.65 mL/L/h, R_max(MGM) of 31.90 mL/L/h, λ of 13.92 h and R² of 0.9999 were obtained at the optimum pH 8. At the optimum fermentation temperature (35 °C), λ of 4.12 h was achieved to maximize hydrogen production to 828.33 ±21.67 mL/L with R_max(MGM) of 27.98 mL/L/h, and R² of 0.9594. At the optimum inoculum size (10%, v/v), the recorded H_max was 1670.00 ±40.41 mL/L with R_max (Exp) of 60.00 ±2.04 mL/L/h and λ of 13.50 hrs. These findings suggest using E. marmotae RM122 as a prospective biohydrogen producer from cheap agro-industrial wastes.