Batch and continuous extraction of bromelain enzyme by reversed micelles

Abstract

The main aim of this study was to optimize the conditions for bromelain extraction by reversed micelles from pineapple juice (Ananas comosus). The purification was carried out in batch extraction and a micro-column with pulsed caps for continuous extraction. The cationic micellar solution was made of BDBAC as a surfactant, isooctane as a solvent and hexanol as a co-solvent. For the batch process, a purification factor of 3 times at the best values of surfactant agent, co-solvent and salt concentrations, pH of the back and forward extractions were, 100 mM, 10% v/v, 1 M, 3.5 and 8, respectively. For the continuous operation, independent variables optimal point was determined: ratio between light phase flow rate and total flow rate equal to 0.67 and 1 second for the time interval between the pulses. This optimal point led to a productivity of 1.29 mL/min and a purification factor of 4.96.

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).…”

The effect of pH on bromelain partition from Ananas comosus by PEG4000/phosphate ATPS

“…The filtrate contained the bromelain enzyme. Samples were frozen at -5 o C (Fileti et al, 2009 and2010;Fisher et al, 2007;Silveira et al, 2009). Bromelain activity was measured by hydrolysis of a 2% (w/w) casein solution at pH 7.5 and 37 o C for 10 min.…”

“…The amount of soluble peptides in TCA was determined by measuring the absorbance at 280 nm. The method defines one unity of enzyme activity as the amount of enzymes that modifies by 1.0 the absorbance at 280 nm (Baldini et al, 1993;Fileti et al, 2009 and2010;Fisher et al, 2007;Lopes et al, 2007 and2009;Murachi, 1976).…”

“…Total protein and bromelain activity in samples of bottom and top phases were determined by the Bradford method (Bradford, 1976) and bromelain activity method (Baldini et al, 1993;Fileti et al, 2009 and2010;Fisher et al, 2007;Murachi, 1976). The partition coefficient was obtained with Equation 1.…”

“…On optimal condition of batch purification a 3-folds of enzyme recovery was obtained and, a purification factor of 6-folds was found for the continuous ATPS operating at 0.5 for top/total flow and 2 s for the time interval between the pulses (Fileti et al, 2009 and2010;Fisher et al, 2007). A study of bromelain partitioning in twophase aqueous systems containing PEO-PPO-PEO block copolymers was made an value 1.25 was the maximum purification factor found for the bromelain recovery from pineapple fruit via twophase aqueous extraction (Rabelo et al, 2004).…”