Abstract
In this work, a model describing the complete enzyme catalysed synthesis of N-acetylneuraminic acid (Neu5Ac) from N-acetyl-d-glucosamine (GlcNAc) is presented. It includes the combined reaction steps of epimerisation from GlcNAc to N-acetyl-d-mannosamine (ManNAc) and the aldol condensation of ManNAc with sodium pyruvate yielding Neu5Ac. The model is expedient to predict the reaction course for various initial and feed concentrations and therefore to calculate reaction times and yields. The equilibrium constants calculated from the kinetic constants via the Haldane relationship correspond with experimental values very well (0.26 calculated and 0.24 experimental value for the epimerisation, 27.4 l mol−1 calculated and 28.7 l mol−1 experimental for the aldol condensation). The actual relevance of the model is shown by a scale-up. Using the model, an optimisation of reaction conditions in consideration of different targets is possible. Exemplarily, it is presented how the optimal ratio of the two enzymes in the reaction can be determined and how the composition of the reaction solution in a fed-batch reactor can be designed to meet downstream processing needs.
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Acknowledgements
We would like to thank Dr. W. Ruth for performing as well as interpreting the LC-MS measurements. This work was supported by the scholarship programme of the German Federal Environmental Foundation (Deutsche Bundesstiftung Umwelt).
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Zimmermann, V., Hennemann, HG., Daußmann, T. et al. Modelling the reaction course of N-acetylneuraminic acid synthesis from N-acetyl-d-glucosamine—new strategies for the optimisation of neuraminic acid synthesis. Appl Microbiol Biotechnol 76, 597–605 (2007). https://doi.org/10.1007/s00253-007-1033-6
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DOI: https://doi.org/10.1007/s00253-007-1033-6