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Hydrogen and carbon dioxide fermentation to methane, called bio-methanation, is a promising way to provide renewable and easy-to-store energy. The main challenge of bio-methanation is the low gas-to-liquid transfer of hydrogen. Gas injection through a porous membrane can be used to obtain microbubbles and high gas-to-liquid transfer. However, the understanding of bubble formation using a membrane in the fermentation broth is still missing. This study focused on the impact of liquid pressure and flow rate in the membrane, gas flow rate, membrane hydrophobicity, surface, and pore size on the overall gas-to-liquid mass transfer coefficient (KLa) for hydrogen with gas injection through a porous membrane in real fermentation conditions. It has been shown that KLa increased by 13% with an increase in liquid pressure from 0.5 bar to 1.5 bar. The use of a hydrophilic membrane increased the KLa by 17% compared to the hydrophobic membrane. The membrane with a pore size of 0.1 µm produced a higher KLa value compared to 50 and 300 kDa. The liquid crossflow velocity did not impact the KLa in the studied range.