The importance of a chemical that helps fight against Influenca is, in times of a global pandemic, self-evident. Many researchers see the continuous production in chemical industry as its next stepping stone. For these reasons, the synthesis of N-Acetylneuraminic acid in a continuous fixed-bed reactor by an immobilized epimerase and aldolase was investigated in detail. The immobilized enzymes showed high stability with half-life times > 173 days under storage condition (6 °C in buffer) and reusability over 50 recycling steps and was characterized regarding the reaction kinetics (initial rate) and scalability (different lab scales) in a batch reactor. In a continuous flow reactor reaction kinetics were studied. A high pressure circular reactor (up to 130 MPa) was applied for investigation of changes in the position of the reaction equilibrium. By this, equilibrium conversion, selectivity and yield were increased from 57.9% to 63.9%, 81.9% to 84.7% and 47.5% to 54.1%, respectively. This indicates a reduction in molar volume from GlcNAc and Pyr to Neu5Ac. The circular reactor in particular showed great potential to study reactions at high pressure while allowing for easy sampling. Additionally, an increase in affinity of pyruvate towards both tested enzymes was found with the application of pressure as measured in a decrease of for the epimerase and for the aldolase of 108 to 42 mM and 91 to 37 mM, respectively.
Keywords: High Pressure; Neu5Ac; GlcNAc; ManNAc; Pyruvate; Immobilization; Enzyme; Epimerase; Aldolase; Continuous Fixed Bed Reactor
When a peer-reviewed version of this preprint is available, this information will be updated in the information box above. If no peer-reviewed version is available, please cite this preprint using the following information:
Reich, J. A.; Aßmann, M.; Hölting, K.; Bubenheim, P.; Kuballa, J.; Liese, A. Beilstein Arch. 2022, 202211. doi:10.3762/bxiv.2022.11.v1
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