Affinity based reverse micellar extraction and separation (ARMES) technique is employed for the first time to extract and purify bromelain from pineapple (Ananas comosus L Merry!) waste. The reverse micellar system of cetyltrimethylammonium bromide (CTAB)/isooctane/butanol/hexanol and sodium bis(2-ethyl-1-hexyl) sulfosuccinate (AOT)/isooctane is used for forward and reverse extractions respectively, with concanavalin A (Con A) as an affinity ligand. The effect of various process parameters like concentration of Con A during forward extraction, type of counter ligand and its concentration during back extraction, type of surfactants and its concentration, aqueous phase pH for reverse extraction on the purification and activity recovery of bromelain has been studied in detail. The optimized conditions of extraction resulted in purification of 12.32 fold with an activity recovery of 185.6%, which is higher than that reported for conventional reverse micellar extraction (RME). The forward, back, reverse and overall extraction efficiencies were found to be 49%, 44%, 48% and 14% respectively under optimized conditions. (C) 2011 Elsevier Ltd. All rights reserved.