Dr.-Ing. Florian Golombek – Dynamic kinetic resolution

The dynamic kinetic resolution of racemates is a powerful method for the production of enantiopure compounds. However, it is not uncommon to face incompatibilities between the cleavage and racemization reactions. Kinetic resolutions are often catalyzed by lipases via selective acylation. Unfortunately, their combination with racemases often turns out to be critical, due to the fact that such acylations normally are run in organic solvents with low water activity. In such solvents most racemases are inactive making a spatial separation of the two reactions mandatory.

Symbol picture of a vesicle - nonbioreactor (sphere) filled with proteins.

The aim of this project was to develop a new type of nano-scale enzyme membrane reactors for the dynamic kinetic resolutions in biphasic systems. To achieve this, polymer-based vesicles, so-called polymersomes, were used as nano-reactors with soluble enzymes encapsulated inside. To avoid disintegration by the organic solvents in the biphasic systems, the membranes of the polymersomes were stabilized. Additionally, the selective mass transport over the membrane was facilitated via  the insertion of appropriate channel proteins. After the establishment of the complete nano-reactor-based biphasic reaction system, the resulting synthesis was thoroughly characterized and a model-supported optimization was performed.

This project was funded by the Federal Ministry of Education and Research (Basistechnologien Nachwuchsgruppe: UniComp – Funktionelle Erweiterung polymerbasierter Reaktionskompartimente für Multienzymsynthesen, project number 031B0221).

Publications:

Golombek F (2022): Deracemisierungen mit Enzym-Membranreaktoren im Nanomaßstab. Dissertation.

Golombek F, Haumann M, Knoll M, Fröba AP, Castiglione K (2021): Three Steps, Two Enzymes, One Pot, but a Multitude of Nanocompartments: Combined Cycles of Kinetic Resolutions and Re-racemization with Incompatible Biocatalysts. ACS Omega, 6, 43, 29192–29200.

Mertz M, Golombek F, Boye S, Moreno S, Castiglione K (2021): Fast and effective chromatographic separation of polymersomes from proteins by multimodal chromatography. J Chrom B, 1162, 122459.

Golombek F, Castiglione K (2020): Polymersomes as Nanoreactors Enabling the Application of Solvent-Sensitive Enzymes in Different Biphasic Reaction Setups. Biotechnol J, 15, 1900561.