Expanded bed adsorption of bromelain (E.C. 3.4.22.33) from Ananas comosus crude extract

Abstract

-This work focuses on the adsorption of Bromelain in expanded bed conditions, such as the adsorption kinetics parameters. The adsorption kinetics parameters showed that after 40 minutes equilibrium was achieved and maximum adsorption capacity was 6.11 U per resin mL. However, the maximum adsorption capacity was only determined by measuring the adsorption isotherm. Only by the Langmuir model the maximum adsorption capacity, Qm, and dissociation constant, kd, values could be estimated as 9.18 U/mL and 0.591, respectively, at 25 °C and 0.1 mol/L phosphate buffer pH 7.5. A column made of glass with an inner diameter of 1 cm was used for the expanded bed adsorption (EBA). The residence time was reduced 10 fold by increasing the expansion degree 2.5 times; nonetheless, the plate number (N) value was reduced only 2 fold. After adsorption, the bromelain was eluted in packed bed mode, with a downward flow. The purification factor was about 13 fold and the total protein was reduced 4 fold. EBA showed to be feasible for purification of bromelain.

Bromelain: an overview of industrial application and purification strategies

“…As bromelain has the property of having a single oligosaccharide chain attached to the polypeptide, Gupta and Saleemuddin (2006) used affinity binding to a Sepharose matrix pre-coupled with the lectin concanavalin A (Con A), which resulted in 60 % bromelain activity at pH 11. Meanwhile, Silveira et al (2009) integrated the expanded bed adsorption (EBA) to investigate the effect of the expansion degree of Amberlite IRA 410 resin and the residence time distribution, which resulted in 13-fold bromelain purification. Recently, the use of dye ligand affinity membrane chromatography seems more promising because it offers high selectivity, a larger surface area, and excellent thermal and mechanical properties.…”

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.…”

“…Besides the pharmacological effects, bromelain is also employed in food industries, such as breweries and meat processing. The optimum pH and temperature of bromelian are 6.5-7.5 and 37°C, respectively (Fisher et al, 2007;Lopes et al, 2007 and2009;Silveira et al, 2009).…”

“…It is difficult and expensive to selectively recover a targeted protein from a broth due to the low protein concentration and the similarity of the physical properties between the proteins present in the same solution. Aqueous two-phase systems (ATPS), expanded bed adsorption (EBA) and membrane separation process are more recent downstream processes used in the purification of biomolecules (Biazus et al, 2006(Biazus et al, , 2007(Biazus et al, and 2010Curvelo-Santana et al, 2008;Fisher et al, 2007;Lopes et al, 2007 and2009;Padilha et al, 2009;2009;Silveira et al, 2009;Toledo et al, 2007).…”

Isolation and purification of bromelain from waste peel of pineapple for therapeutic application

“…To achieve the required characteristics for pharmaceutical usage, many approaches have been used to increase the purity and specific activity of bromelain from pineapple peel, stem and fruit. These include liquid-liquid extraction procedures (Rabelo et al 2004;Badu 2008;Ketnawa 2010;Ferreira et al 2011), ethanol, polyethyleneglycol or ammonium sulphate precipitation (Singh 2004;Devakate et al 2009;Silva et al 2010;Gautam et al 2010;Soares et al 2011), and polishing techniques, such as ion exchange and affinity chromatography (Harrach et al 1998;Singh 2004;Devakate et al 2009;Silveira 2009;Gautam et al 2010), followed by freeze-drying for storage ends (Maurer 2001;Devakate et al 2009). In the literature, however, there is a lack of protocols aiming at bromelain recovery and purification from by-products of food industry such as pineapple peel.…”