Two-phase partitioning bioreactors (TPPBs) in environmental biotechnology are based on the addition of a non-aqueous phase (NAP) into a biological process in order to overcome both mass-transfer limitations from the gas to aqueous phase and pollutant-mediated inhibitions. Despite constituting a robust and reliable technology in terms of pollutant biodegradation rates and process stability in wastewater, soil, and gas treatment applications, this superior performance only applies for a restricted number of pollutants or contamination events. Severe limitations such as high energy requirements, high costs of some NAPs, foaming, or pollutant sequestration challenge the full-scale application of this technology. The introduction of solid NAPs into this research field has opened a promising pathway for the future development of TPPBs. Finally, this work reviews fundamental aspects of NAP selection and mass transfer and identifies the niches for future research: low energy-demand bioreactor designs, experimental determination of partial mass transfers, and solid NAP tailoring.