Glycosyltransferases (GTs) Engineering Services

Glycosyltransferases (GTs) are fundamental to the synthesis of complex carbohydrates, playing an increasingly vital role in Glycoprotein and Glycopeptide Drug Modification. Precise glycosylation is essential for drug efficacy, half-life, and immunogenicity. However, utilizing GTs for synthetic glycosylation faces critical challenges: the low availability and high cost of natural sugar nucleotide donors, poor enzymatic transfer efficiency, and the inherent difficulty in achieving complex, non-natural glycosylation patterns that enhance therapeutic profiles.

Our specialized enzyme optimization services and powerful engineering platforms are dedicated to addressing these biochemical and cost barriers. Our focus areas include: engineering GTs for broader sugar donor promiscuity (utilizing cheaper substrates), significantly improving transfer efficiency and regioselectivity, and creating novel GT variants to synthesize highly specific and complex glycan structures. Consult with our experts to design a customized strategy that elevates your glycosylation process to an industrial and therapeutic standard.

Challenges in Glycosyltransferase (GT) Biocatalysis

The large-scale, cost-effective utilization of GTs for drug modification is limited by these key factors:

• Expensive Sugar Nucleotide Donors: GTs typically require activated sugar donors (e.g., UDP-sugars), which are difficult and costly to synthesize or obtain, posing a major barrier to scale-up.
• Low Catalytic Efficiency: Many GTs, particularly those involved in complex pathways, exhibit low transfer rates and poor conversion, leading to lengthy reaction times.
• Limited Substrate Promiscuity: Natural GTs are highly specific, restricting their application to synthesize non-natural or novel glycoconjugates required for next-generation therapeutics.
• Regio/Stereoselectivity Control: Achieving precise linkage specificity (e.g., \alpha-1,2 vs. \alpha-1,3) or selectively glycosylating one of multiple hydroxyl groups remains a major challenge.

Our engineering platforms are dedicated to resolving these complex stoichiometric and specificity limitations.

Engineering Focus: Cost Reduction and Glycan Precision

We apply integrated protein engineering strategies to enhance your target Glycosyltransferase:

Sugar Donor Promiscuity Expansion

             

We engineer the sugar donor binding pocket for increased tolerance, enabling the use of cheaper, more accessible activated sugar precursors.

Transfer Efficiency Improvement

Advanced enzyme optimization services focusing on improving k_cat and coupling the reaction with efficient sugar donor regeneration systems.

Regio- & Glycan Specificity Control

We use specificity engineering to achieve precise glycan linkage and to create novel GTs that synthesize specific, complex non-natural glycan structures.

Integrated Cascade Design

We design multi-enzyme cascade systems, combining GTs with donor regeneration enzymes for a streamlined, single-pot synthesis of glycosylated products.

Our experts are ready to apply these integrated capabilities to your specific glycoprotein or glycopeptide modification project.

Technology Platforms for Glycosyltransferase Engineering

We leverage a suite of cutting-edge platforms to deliver highly functional enzyme variants:

Computational Rational Design (CARD)

Using substrate interaction modeling and structural bioinformatics, we guide mutations in the donor/acceptor binding sites to predict changes in promiscuity and specificity.

Directed Evolution Workflows

We utilize HTS services coupled with sensitive glycosylation assays (e.g., coupled-enzyme assays) to rapidly evolve GTs for enhanced efficiency and non-natural substrate use.

Cell Surface Display for Engineering

Platforms like Yeast Surface Display are employed to screen large libraries of GT variants, particularly those requiring membrane localization, for improved activity or stability.

Comprehensive Enzyme Profiling

We offer full enzyme characterization, including kinetic analysis of both sugar donor and protein acceptor, and stability profiling under process conditions.

Optimized Enzyme Production

Specialized custom production services to achieve high yield and purity for membrane-bound or complex GTs in appropriate expression systems.

Partner with us to harness these platforms for your next glycobiology breakthrough.

Project Flow: GT Optimization Workflow

Our enzyme optimization projects follow a flexible, milestone-driven workflow:

• Consultation and Goal Definition: Initial discussion to define precise transfer efficiency, specificity (linkage type), and donor promiscuity targets.
• Design Strategy Proposal: We propose a tailored strategy involving Rational Design, Directed Evolution, and/or cascade integration, outlining the predicted timeframe and resources.
• Library Construction and Screening: We execute mutagenesis and employ HTS platforms to identify lead variants that meet intermediate efficiency and specificity milestones.
• Iterative Optimization & Profiling: Successive rounds of evolution are performed under increasingly challenging conditions (e.g., using low-cost donors) to build industrial robustness.
• Final Deliverables: Delivery of the final GT enzyme variant along with detailed kinetic, specificity, and stability reports ready for therapeutic application or scale-up synthesis.

Technical communication is maintained throughout the project. We encourage potential clients to initiate a consultation to discuss their specific GT requirements and explore how our technologies can achieve their desired glycan structures and process cost reduction.

We provide comprehensive support, including:

• Detailed Kinetic Data, Linkage Specificity, Acceptor/Donor Promiscuity, and Transfer Efficiency Reports.
• Consultation on process integration, including the design of efficient in-situ sugar nucleotide regeneration systems.
• Experimental reports include complete raw data on mutagenesis libraries, screening results, and glycan analysis (e.g., HPLC/Mass Spec) traces.