Purification of chimeric amine dehydrogenase using a tailor-made aqueous two-phase system - A case study

Enzymes used as biocatalysts in evolving green processes have already reached industrial competitiveness to classical chemical catalysts. After production of these enzymes, aqueous two-phase extraction using an aqueous two-phase system (ATPS) has been shown a promising and sustainable alternative to common cost-intense chromatographic purification. However, state-of-the-art ATPS design is based on trial-and-error screening approaches leading to un-optimized ATPS. Within this work, we demonstrate the applicability of our method for ATPS design to an industrially relevant chimeric amine dehydrogenase (cFL1). The method includes thermodynamic modeling of ATPS phase composition and investigations on the conformational and colloidal cEL1 stability to rapidly select a tailor-made ATPS, offering optimal process and processing conditions, outclassing the previously selected ATPS (identified by random screening) by a factor of 4. All investigations and the selection of the sodium glutamate-polyethylene glycol 2000 ATPS were performed in less than 4 weeks, demonstrating proof of concept for a fast and sustainable ATPS selection in an early stage of downstream processing development. (C) 2020 Elsevier B.V. All rights reserved.

Automated summary

Enzymes used as biocatalysts have become competitive with classical chemical catalysts in green processes, and aqueous two-phase extraction has been shown as a promising and sustainable alternative for purification. A new method for ATPS design has been demonstrated to rapidly select tailor-made ATPS, outperforming traditional trial-and-error screening approaches.