Purification, enzymatic activity and inhibitor discovery for recombinant human carbonic anhydrase XIV

•Recombinant preparation of CA XIV, a difficult to produce protein, is described.•Large amount of CA XIV was affinity purified using sulfonamide-modified sepharose.•Inhibitor titration of activity demonstrated nearly 100% pure and active enzyme.•Nanomolar inhibitors of CA XIV, selective over CA I and II, were designed.