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Trends in Chromatography   Volumes    Volume 3 
Aqueous two-phase systems chromatography and biochemistry characterization of a enzyme from Ananas comosus
Gilvan Anderson Fischer, Juliana Ferreira Ferrari, José Carlos Curvelo Santana, Ana Maria Frattini Fileti, Elias Basile Tambourgi, Juliana de Moura Luna
Pages: 85 - 95
Number of pages: 11
Trends in Chromatography
Volume 3 

Copyright © 2007 Research Trends. All rights reserved

This work aimed to purify and characterize the bromelain from pineapple juice (Ananas comosus) by batch and continuous extraction by reversed micelles ATPS. The cationic micellar solution was made up of BDBAC as a surfactant, isooctane as a solvent and hexanol as a co-solvent. A micro-column with pulsed caps was used in the continuous extraction. Batch ATPS purification concentrated the enzyme in 3 times and the best values of surfactant agent, co-solvent and salt concentrations, pH of the back and forward extractions were, respectively: 100 mM, 10% v/v, 1 M, 3.5 and 8. A purification factor of 6 times was found for the continuous ATPS operating at 0.5 for top/total flow and 2 second for the time interval between the pulses. Enzyme characterization showed that pH 7 was the optimal pH, the optimal temperature were finding between 30-40°C, the Km and Vmax values were of 3.30 10-2 mg/mL and 2.54 mg/mL.min, respectively and SDS-PAGE electrophoresis showed that the bromelain from Ananas comosus pulp had a molecular mass of 24.5 kDa.
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