Chemoenzymatic production of bioorthogonal carbohydrates for glycan research applications

The National Institute of General Medical Sciences awarded Midwest Bioprocessing Center, Inc. $313,957 on August 13, 2026, under the Biomedical Research and Research Training program (CFDA 93.859) to develop bioorthogonal nucleotide sugars for glycan research applications using chemoenzymatic production methods.

The Phase I award funds demonstration of production of three nucleotide sugars containing bioorthogonal handles through enzymatic pathways. The work encompasses synthesis of initial sugars with bioorthogonal handles, enzymatic transformation of modified sugars into sugar-1-phosphates, and synthesis of model bioorthogonal nucleotide sugars from the resulting sugar-1-phosphates using enzymatic means. Success will demonstrate proof of concept for the chemoenzymatic process, establishing feasibility for Phase II optimization toward gram-scale or greater production. Bioorthogonal chemistry enables chemical reactions that do not disrupt biological systems in physiological environments; applying these techniques to nucleotide sugars will expand commercially available bioorthogonal reagents for drug discovery, cancer research, host-pathogen interaction studies, cell signaling research, and assay development.

Performance occurs in Peoria, Illinois, with completion targeted for July 31, 2027.

Abstract

The goal of this project is to develop a series of bioorthogonal nucleotide sugars that can be used in drug discovery research applications using a chemoenzymatic strategy. Bioorthogonal chemistry involves the utility of chemical reactions that do not disrupt biological systems in a physiological environment. Bioorthogonal chemistry techniques have seen great success when applied to biomolecules within many topics of research. Glycan research is one research area that has benefitted greatly from bioorthogonal chemistry. By using reporter molecules, visualization and manipulation of glycans has advanced research in the fields of cancer/oncology, host-pathogen interactions, cell signaling, and assay development. As the interest in and usage of bioorthogonal techniques increases, the need for additional commercially available bioorthogonal handles becomes greater. Utilizing our chemoenzymatic technology, we propose to investigate the development of additional nucleotide sugars that contain bioorthogonal handles. Achieving this will require testing our enzymes for their ability to tolerate non-native substrates. Our phase I plan is to demonstrate production of three nucleotide sugars that utilize different enzymatic pathways. This will be accomplished by achieving three goals: 1) synthesizing the initial sugars with their bioorthogonal handles, 2) demonstrating the enzymatic transformation of modified sugars into sugar-1- phosphates, and 3) demonstrating the synthesis of model bioorthogonal nucleotide sugars from the sugar-1- phosphates using enzymatic means. We ideally will show success in the production of at least one target molecule. Completion of phase I will show that our chemoenzymatic process is working and can be tested on an expanded substrate scope. Phase II will initiate the optimization in production of these molecules to achieve gram scale or greater, and to expand the product scope of bioorthogonal nucleotide sugars we can achieve. Phase III will involve commercialization of our developed products to increase their availability in the scientific community.