Modelling approach for the continuous biocatalytic synthesis of N-acetylneuraminic acid in packed bed reactors

Journal
Processes  
Volume
12
Issue
10
Article Number
2191
Citation
Processes 12 (10): 2191 (2024)
Publisher DOI
10.3390/pr12102191
Scopus ID
2-s2.0-85207448489
Publisher
Multidisciplinary Digital Publishing Institute
Continuous flow technologies have become increasingly important for biocatalytic processes. In this study, we present the application and modelling of covalently immobilised <i>N</i>-acetylglucosamine 2-epimerase and <i>N</i>-acetylneuraminic acid lyase in packed bed reactors for the synthesis of <i>N</i>-acetylneuraminic acid. The immobilised enzymes were stable under continuous flow process conditions with half-life times of >28 d (epimerase immobilised on hexamethylamino methacrylate HA403/M) or 58 d (lyase immobilised on dimenthylamino methacrylate ECR8309M), suitable for continuous flow applications. Kinetic studies revealed Michaelis–Menten kinetic behaviour for both enzymes. The kinetic parameters and the inhibitions were analysed under continuous flow conditions and were integrated into a process model using Python. The model was validated by varying flow rates, the mass of immobilised enzymes and the reactor dimensions and shows a low error compared to the measured data. An error accuracy of 6% (epimerase) or 9% (lyase) was achieved. The product concentrations of the enzyme cascade at the end of the packed bed reactor can be predicted with an accuracy of 9% for the calculation of a large column (84.5 mL) or of 24% if several small columns (2.5 mL, 0.8 mL) are connected in series. The developed model has proved to be valid and will be used to optimise the process with respect to substrate concentrations, reactor dimensions and flow rate.
Subjects
continuous biocatalysis
flow tube
GlcNAc 2-epimerase
immobilisation
N-acetylneuraminic acid
Neu5Ac lyase
packed bed reactor
DDC Class
570: Life Sciences, Biology
610: Medicine, Health
660: Chemistry; Chemical Engineering
620: Engineering
Lizenz
https://creativecommons.org/licenses/by/4.0/
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