Abstract
The objective of this work was a proof-of-concept for the use of the immobilized bovine α-chymotrypsin enzyme in a biotransformation process applying an aqueous micellar two-phase system (AMTPS) for separation and recycling of the enzyme immobilisates. Commercially available magnetic polyvinyl alcohol microparticles serve as carriers for the covalent immobilization of the enzyme. For the AMTPS non-ionic surfactants are used, creating a temperature controlled two-phase system which allows for an easy separation of the immobilized enzyme from the reaction medium. We describe the influence of four non-ionic surfactants (Tween 20, Triton X 114, Triton X 100, and Eumulgin ES) on the activity of the immobilized α-chymotrypsin and the optimization of the AMTPS composition via a Design of Experiments approach. Additionally, we demonstrate the reuse of the immobilisates for hemoglobin digestion over eleven cycles in an Eumulgin ES AMTPS. Our findings show that continuous enzymatic conversion processes can be implemented effectively with the help of carrier bound enzymes in surfactant systems.
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