Monday, May 5, 2008
9-31

Two-Phase Aqueous Micellar Systems in Oscillatory Flow Micro-Reactor: Study of Perspectives and Experimental Performance

Daniel P. Silva1, André M. Lopes2, António A. Vicente3, Adalberto Pessoa Jr.2, and José A. Teixeira3. (1) IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar, Braga, 4710-057, Portugal, (2) Department of Biochemical-Pharmaceutical Technology, School of Pharmaceutical Sciences, Universidade de São Paulo, São Paulo, 05508-900, Brazil, (3) IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, Braga, 4710-057, Portugal

Recently several optimizations for the extraction/purification of biological products have been reported. Among the techniques used, aqueous two-phase micellar systems (ATPMS) are micellar surfactants solutions with physical properties that make them very efficient (e.g. excluded-volume interactions). However, in this work, we report our search for alternative methods and/or development of new procedures in extraction systems, as the application of a novel oscillatory flow micro-reactor (micro-OFR). This batch or continuous micro-reactor based on oscillatory flow mixing (OFM) technology has been presented as a new small-scale alternative for biological or physical-chemical applications. OFRs can be operated continuously in horizontal or vertical tubes, and their mechanical conditions can be linearly scaled up. Thus, in this work ATPMS experiments were carried out in different OFM conditions (temperatures, oscillation frequencies and amplitudes) for the extraction of glucose-6-phosphate dehydrogenase (G6PD) in TritonX-114/buffer with Cibacron Blue as affinity ligand. The phase separations were obtained by resting under different temperatures. The G6PD activity was obtained and the partition coefficient calculated. Although, at the different applied conditions no improvements were observed in the partitioning selectivity of the G6PD using ATPMS with affinity ligand when compared with those obtained in simple rotation (KG6PD~0.3), the results suggest that the micro-OFR technology can be considered a promising and new alternative for screening studies in extraction systems or industrial bioprocesses. However, the different conditions applied during this work were not enough to improve, through affinity interactions, the partitioning selectivity of the target enzyme.
Acknowledgements: FCT (Portugal) and FAPESP (Brasil)

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