In this paper, pectin-reinforced, supported liquid membranes (SLMs) prepared with carbonic anhydrase (CA) were investigated for CO2/N2 separation. In the first part of the study, the effect of [Bmim][NTf2] ionic liquid (IL) – as possible solvent to fill the pores of cellulose acetate support during SLM fabrication – on enzyme activity was tested. It turned out that this particular IL caused rapid and severe loss of initial biocatalyst activity, which fact can be seen as a threat in the membrane process design. Afterwards, the stability of pectin-containing SLMs (containing CA but lacking the IL having adverse impact) was addressed and their improved resistance against higher transmembrane pressures (up to 7.2 bar) was found, representing an approx. 3-fold enhancement compared to their control. Thereafter, the performance of the membranes was tested under single and mixed gas conditions with carbon dioxide and nitrogen. Employing single gases, it was demonstrated that CA enzyme could notably increase CO2 permeability (from 55 to 93 Barrer), while that of N2 remained unchanged (1.6-1.7 Barrer). Thus, the highest CO2/N2 theoretical selectivity was attained as 54 using the pectin-reinforced SLMs enriched with CA biocatalyst. For comparison, the outcomes were plotted on the Robeson upper-bound.