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Biocatalytic synthesis routes offer unique advantages over classical chemical processes. In general, they operate under mild conditions offering high reaction rates and selectivity, making them of great interest in the development of sustainable production processes [1]. Besides innovation in the upstream process, research effort has been spent in improving the efficiency and sustainability of the following purification step. Aqueous Two-Phase Systems (ATPS) are promising alternatives to organic solvents due to their gentleness toward biomolecules, their ability to offer high partition coefficients and yields, and their ease of scale-up [2]. In this work, the advantages of both approaches will be combined in a reactive ATPS with a biocatalytic reaction therein.
The main goal of this work is to design and model reactive ATPS using the equation of state ePC-SAFT advanced [3]. Besides this engineering goal, liquid-liquid two-phase (LLTPS) separation causing coacervate formation is observed in life cells that use reactions in such compartments for the sake of spatio‑temporal organization. Coacervates are liquid droplets enriched in macromolecules built by specific interactions between their building blocks. The LLTPS constituents and the operating conditions (temperature, pressure, pH) will be tuned to maximize yield, selectivity, and the partitioning of substrates and products.
Figure 1: Illustration of the enzymatic reaction network underlying formation/dissolution of the ATP/pLys-coacervate droplets (HK: hexokinase, PK: pyruvate kinase) [4].
