Gas transfer and mixing were characterized in a 32-L bubble column reactor equipped with a commercially available rubber membrane diffuser. The performance of the membrane diffuser indicates that the slits in the membrane are best described as holes with elastic lids, acting as valves cutting off bubbles from the gas stream. The membrane diffuser thus functions as a one-way valve preventing backflow of liquid. Our design of the bottom plate of the reactor enabled us to optimize the aeration by changing the tension of the membrane. We thereby achieved mass transfer coefficients higher than those previously reported in bubble columns. A strong dependence of mass transfer on gas holdup and bubble size was indicated by estimates based on these two variables. The microalga, Rhodomonas sp., sensitive to chemical and physical stress, was maintained for 8 months in continuous culture with a productivity identical to cultures grown in stirred tank reactors.