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Dark fermentation for bio-hydrogen (bio-H2) production is an easily operated and environmentally friendly technology. However, low bio-H2 production yield has been reported as its main drawback. Two strategies have been followed in the past to improve this fact: genetic modifications and adjusting the reaction conditions. In this paper, the second one is followed to regulate the bio-H2 release from the reactor. This operating condition alters the metabolic pathways and increased the bio-H2 production twice. Gas release was forced in the continuous culture to study the equilibrium in the mass transfer between the gaseous and liquid phases. This equilibrium depends on the H2, CO2, and volatile fatty acids production. The effect of reducing the bio-H2 partial pressure (bio-H2 pp) to enhance bio-H2 production was evaluated in a 30 L continuous stirred tank reactor. Three bio-H2 release strategies were followed: uncontrolled, intermittent, and constant. In the so called uncontrolled fermentation, without bio-H2 pp control, a bio-H2 molar yield of 1.2 mol/mol glucose was obtained. A sustained low bio-H2 pp of 0.06 atm increased the bio-H2 production rate from 16.1 to 108 mL/L/h with a stable bio-H2 percentage of 55 % (v/v) and a molar yield of 1.9 mol/mol glucose. Biogas release enhanced bio-H2 production because lower bio-H2 pp, CO2 concentration, and reduced volatile fatty acids accumulation prevented the associated inhibitions and bio-H2 consumption.