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Pancreatic serine protease extraction by affinity partition using a free triazine dye
Maria Victoria Rocha1, Diana Romanini, Bibiana B Nerli
1Departamento Química-Física, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.
International Journal of Biological Macromolecules
|January 3, 2012
Summary
Reactive Yellow 2 (RY2) acts as a biomimetic ligand for trypsin (TRP) and chymotrypsin (ChTRP). This low-cost dye enhances their separation in aqueous two-phase systems, demonstrating its potential for affinity partitioning.
Area of Science:
- Biochemistry
- Separation Science
Background:
- Affinity partitioning leverages biorecognition in aqueous two-phase systems for macromolecule separation.
- Reactive dyes are cost-effective biomimetic ligands for various enzymes.
Purpose of the Study:
- To investigate the interaction of Reactive Yellow 2 (RY2) with trypsin (TRP) and chymotrypsin (ChTRP).
- To evaluate RY2's behavior and efficacy as an affinity ligand in polyethylene glycol (PEG)/sodium citrate (NaCit) aqueous two-phase systems.
Main Methods:
- Investigated RY2's binding affinity and partition behavior in PEG/NaCit systems.
- Analyzed effects of PEG molecular weight, tie line length, and dye concentration.
- Quantified changes in TRP and ChTRP partition coefficients with RY2 addition.
Main Results:
- RY2 specifically binds to TRP and ChTRP with affinity constants around 10(3)M(-1).
- RY2's partition equilibrium favors the top phase in PEG-based systems.
- RY2 addition increased TRP and ChTRP partition coefficients by at least two-fold.
Conclusions:
- RY2 demonstrates specific binding and effective partitioning behavior for TRP and ChTRP.
- RY2 fulfills the criteria for an effective affinity ligand in aqueous two-phase partitioning of these enzymes.
