Abstract
Aqueous two-phase systems (ATPS) have been reported as an attractive biocompatible extraction system for recovery and purification of biological products. In this work, the implementation, characterization, and optimization (operational and economic) of invertase extraction from spent brewery yeast in a semi-automatized pilot plant using ATPS is reported. Gentian violet was used as tracer for the selection of phase composition through phase entrainment minimization. Yeast suspension was chosen as a complex cell matrix model for the recovery of the industrial relevant enzyme invertase. Flow rates of phases did not have an effect, given that a bottom continuous phase is given, while load of sample and number of agitators improved the recovery of the enzyme. The best combination of factors reached a recovery of 129.35 ± 2.76% and a purification factor of 4.98 ± 1.10 in the bottom phase of a PEG-Phosphate system, also resulting in the removal of inhibitor molecules increasing invertase activity as reported by several other authors. Then, an economic analysis was performed to study the production cost of invertase analyzing only the significant parameters for production. Results indicate that the parameters being analyzed only affect the production cost per enzymatic unit, while variations in the cost per batch are not significant. Moreover, only the sample load is significant, which, combined with operational optimization results, gives the same optimal result for operation, maximizing recovery yield (15% of sample load and 1 static mixer). Overall res ults of these case studies show continuous pilot-scale ATPS as a viable and reproducible extraction/purification system for high added-value biological compounds.
Highlights
With the improvement in recombinant technologies, great progress has been taking place in the upstream processes of bioproducts manufacturing
The hydrodynamic parameters of different kinds of extraction columns have been investigated by several researchers but not for aqueous two-phase systems (ATPS) systems in pilot plant scale
Considering two ATPS compositions previously explored for the recovery of enzymes (Vázquez-Villegas et al, 2011, 2015), phase entrainments were compared
Summary
With the improvement in recombinant technologies, great progress has been taking place in the upstream processes of bioproducts manufacturing. Continuous ATPS for Enzyme Recovery operational windows or scaling up drawbacks, besides the use of high cost equipment (Xu et al, 2003; Aguilar et al, 2010; Rosa et al, 2011; Goja et al, 2013) This has promoted the development and implementation of alternative, scalable extraction procedures and purification systems that allow higher throughputs. The study of alternative equipment configurations such as mixer settlers and the development of novel separators for different ATPS variants, bioproducts and physicochemical characteristics could increase the interest toward the industrial implementation of continuous ATPS-based processes (Cuhna and Aires-Barros, 2002; Benavides and Rito-Palomares, 2008; Torres-Acosta et al, 2016). It is based on a tubular reactor approach with a large and adjustable length/diameter ratio to control settling time and separation of phases (Vázquez-Villegas et al, 2011)
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