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Publisher DOI: 10.3762/bjoc.18.59
Title: Shift of the reaction equilibrium at high pressure in the continuous synthesis of neuraminic acid
Language: English
Authors: Reich, Jannis Alexander 
Aßmann, Miriam 
Hölting, Kristin 
Bubenheim, Paul  
Kuballa, Jürgen 
Liese, Andreas  
Keywords: aldolase; continuous fixed-bed reactor; enzyme; epimerase; GlcNAc; high pressure; immobilization; ManNAc; Neu5Ac; pyruvate
Issue Date: 20-May-2022
Publisher: Beilstein-Institut zur Förderung der Chemischen Wissenschaften
Source: Beilstein Journal of Organic Chemistry 18: 567-579 (2022)
Abstract (english): 
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-D-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.
DOI: 10.15480/882.4541
ISSN: 1860-5397
Journal: Beilstein journal of organic chemistry 
Institute: Technische Biokatalyse V-6 
Document Type: Article
Project: Protein Pressure Specific Activity Impact "Modulation der Reaktivität von Proteinen und Thermodynamik durch Druck" 
License: CC BY 4.0 (Attribution) CC BY 4.0 (Attribution)
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