The importance of a compound that helps fight against influenza is, in times of a pandemic, self-evident. In order to produce these compounds in vast quantities, many researchers consider continuous flow reactors in chemical industry as next stepping stone for large scale production. For these reasons, the synthesis of N-acetylneuraminic acid (Neu5Ac) 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 conditions (6 °C in buffer) and reusability over 50 recycling steps, and were characterized regarding the reaction kinetics (initial rate) and scalability (different lab scales) in a batch reactor. The reaction kinetics were studied in a continuous flow reactor. A high-pressure circular reactor (up to 130 MPa) was applied for the 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 N-acetyl-ᴅ-glucosamine (GlcNAc) and pyruvate (Pyr) to Neu5Ac. In particular, the circular reactor 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 observed when high pressure was applied, as evidenced by a decrease of KI for the epimerase and KM for the aldolase from 108 to 42 mM and 91 to 37 mM, respectively.
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.
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