Glycosyltransferase Mutation-Activity Landscape

Background:

Glycosyltransferases are enzymes that commonly transfer monosaccharides to proteins and lipid scaffolds.  These enzymes constitute ~2% of the coding human genome, representing over 200 proteins.  Mutations in glycosyltransferases lead to disease; dysregulated glycosyltransferase expression occurs in metabolic diseases including cancers; modifications of glycosyltransferases offer therapeutic applications; and activity manipulations create novel biocatalysts.  This technology provides a streamlined high-throughput platform for engineering glycosyltransferases and glycosidases.

Technology Overview:

This University of Buffalo technology offers a streamlined platform to perform scanning mutagenesis of arbitrary glycosyltransferases using a novel surface-display platform, creating enzymes with modified activity for biotechnology and therapeutic development. This system allows immobilization of mammalian glycosyltransferase as type-II transmembrane proteins while incorporating a click-chemistry process for quantitating enzyme activity on single cells using natural substrates as acceptors.

Advantages:

• Methods are easy to perform
• Generates large scale data sets needed for modern machine learning
• Economical assay performance
• Generation of new biocatalysts
• Structure-function relationship evaluations

Applications:

• Improving intravenous immunoglobulin (IVIG) therapy
• New enzymes for lysosomal storage disorder (LSD) replacement therapy
• Novel biocatalysts for chemoenzymatic synthesis
• Therapeutic agents for congenital disorders of glycosylation (CDG) diseases
• Customization of biocatalysts for bioengineering process optimization to enhance pharmacokinetics
• Research tools - custom or contract

Intellectual Property Summary:

US Provisional Patent Application 63/812,831 filed on May 27, 2025.

Stage of Development:

Proven application using multiple human sialyltransferases as well as sialyltransferases from species spanning evolutionary progress (fish, sea urchin, frog, monkey, pig, and human).  Creation of a library of 1680 human glycosyltransferases to select for specific desired enzymatic properties, with high activity molecules deemed super-enzymes.  Implementation continues to generate novel molecular species for clinical and commercial applications.

Licensing Status:

Available for licensing or collaboration.

Publications:

• Mammalian Cell Surface Display for High-Throughput Protein Engineering of Glycosyltransferases
• Engineering glycosyltransferases into glycan binding proteins using a mammalian surface display platform | Nature Communications
• Protein engineering strategies to develop lectins by design | Glycobiology | Oxford Academic