The extraction of Penicillin G (Pen G) and its conversion to 6-aminopenicillin acid (6-APA) and phenylacetic acid (PAA) was performed by means of Penicillin G Amidase immobilised in the emulsion liquid membranes. Using various surfactants as emulsifiers and an appropriate carrier it is possible to obtain different extraction rates of Pen G as well as back transfer rates of the hydrolysis products. The surfactants with polyoxyethylene chain facilitate the back transfer through the membrane phase, whereas the more hydrophobic surfactants, e.g., Paranox 100, tend to accumulate Pen G hydrolysis products in the internal aqueous phase.