Process optimization for reverse micellar extraction of stem bromelain with a focus on back extraction

Abstract

In the current study, reverse micellar extraction (RME) for the purification of stem bromelain was successfully achieved using the sodium bis(2-ethylhexyl) sulfosuccinate (AOT)/isooctane system. A maximum forward extraction efficiency of 58.0% was obtained at 100 mM AOT concentration, aqueous phase pH of 8.0 and 0.2 M NaCl. Back extraction studies on altering stripping phase pH and KCl concentration, addition of counter-ion and iso-propyl alcohol (IPA) and mechanical agitation with glass beads indicated that IPA addition and agitation with glass beads have significant effects on extraction efficiency. The protein extraction was higher (51.9%) in case of the IPA (10% v/v) added system during back extraction as compared to a cetyltrimethylammonium bromide (100 mM) added system (9.42%). The central composite design technique was used to optimize the back extraction conditions further. Concentration of IPA, amount of glass beads, mixing time, and agitation speed (in rpm) were the variables selected. IPA concentration of 8.5% (v/v), glass bead concentration of 0.6 (w/v), and mixing time of 45 min at 400 rpm resulted in higher back extraction efficiency of 45.6% and activity recovery of 88.8% with purification of 3.04-fold. The study indicated that mechanical agitation using glass beads could be used for destabilizing the reverse micelles and release of bromelain back into the fresh aqueous phase.

Artificial neural network‐genetic algorithm (ANN‐GA) based medium optimization for the production of human interferon gamma (hIFN‐γ) in Kluyveromyces lactis cell factory

“…The model lack of fit was shown to be insignificant (p > 0.05) with an F value of 0.55. The lack of fit measures the failure of the model to represent data in the experimental domain at points that are not included in the model . Table depicts the model regression co‐efficient that indicates the effect of variables on the hIFN‐γ production.…”

Bromelain: Methods of Extraction, Purification and Therapeutic Applications

“…The use of affinity based reverse micellar extraction and separation technique to extract and purify bromelain from pineapple waste yielded purification of 12.32 fold with an activity recovery of 185.6 % (Kumar et al 2011). Moreover the process optimization studies for RMS had been reported (Fileti et al 2009;Dhaneshwar 2014) and both the batch and continuous extraction of bromelain from pineapple juice by under optimized conditions was also studied (Fileti et al 2007). A neutral model developed for bromelain extraction from pineapple juice by RMS under optimized conditions gave 4.96 purification factor with productivity of 1.29 ml/min (Fileti et al 2009).…”