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A modular, reusable biocatalytic flow system for UDP-GlcNAc production
Tom L Roberts1,2, Jonathan P Dolan1,2, Gavin J Miller1,2
1Lennard-Jones Laboratory, School of Chemical & Physical Sciences, Keele University Keele Staffordshire ST5 5BG UK g.j.miller@keele.ac.uk s.cosgrove@keele.ac.uk.
Reaction Chemistry & Engineering
|May 19, 2025
Summary
Continuous flow synthesis of uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) was achieved using immobilized enzymes in packed bed reactors. This biocatalytic platform offers improved efficiency and scalability for producing valuable glycan targets.
Area of Science:
- Biocatalysis and Chemical Engineering
- Glycobiology and Synthetic Chemistry
Background:
- Sugar nucleotides like uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) are crucial building blocks for synthesizing complex carbohydrates.
- Traditional batch synthesis methods often suffer from low yields, enzyme instability, and product inhibition, limiting scalability.
Purpose of the Study:
- To develop a continuous flow biocatalytic platform for the efficient synthesis of UDP-GlcNAc.
- To improve space-time yield (STY) and overcome limitations of batch processes for sugar nucleotide production.
Main Methods:
- Immobilization of enzymes onto solid carriers for use in packed bed reactors.
- Implementation of a modular continuous flow system allowing tailored conditions for each enzyme.
- Scale-up of UDP-GlcNAc synthesis to the 100 mg level with potential for multiple enzyme reuses.
Main Results:
- Achieved a significant, up to 11-fold, improvement in space-time yield (STY) compared to batch synthesis.
- Prevented product-induced enzyme inhibition and eliminated the need for pyrophosphatase.
- Demonstrated tailored conditions for each enzyme, addressing thermal stability issues.
Conclusions:
- The developed modular continuous flow system provides a highly efficient and scalable platform for UDP-GlcNAc synthesis.
- This approach facilitates the continuous production of important glycan targets, including glycoproteins and glycosylated drugs.
- Represents a significant advancement in biocatalytic synthesis for complex carbohydrate production.
