The effects of three types of surface active agents (containing SDS, HCTBr and Tween 40) with various concentrations (0–5 ppm) on the hydrodynamic and oxygen mass transfer characteristics in a split-cylinder airlift bioreactor with and without packing were investigated. It was observed that in the surfactant solutions, surface tension of the liquid decreased and smaller bubbles were produced in comparison with pure water. So, surfactants presence strongly enhanced mixing time and gas hold-up although oxygen mass transfer coefficient and the liquid circulation velocity reduced. Furthermore, the packing installation enhanced the overall gas–liquid volumetric mass transfer coefficient by increasing flow turbulency and Reynolds number compared to an unpacked column. The packing increased gas hold-up and decreased bubbles size and liquid circulation velocity.