Leyre joined Asparia in 2019 as a Research Biochemist, working in both Glycobiology and Analytical Chemistry. Before that, she gained experience in biopharmaceutical production at a CRO. She specializes in the enzymatic synthesis of complex oligosaccharides and glycopeptides, as well as the production and analysis of synthetic glycans. She’s also involved in N- and O-glycan analysis and the isolation of glycans from natural sources. Leyre holds a Biochemistry degree from the Universidad Autónoma de Madrid and a Master’s in Bioinformatics and Biostatistics from the Universitat Oberta de Catalunya.
In this interview, we’ll be diving into the world of enzymatic synthesis and glycan analysis, breaking down why they’re so essential in biotechnology. We’ll talk about how enzymatic approaches offer significant advantages over traditional chemical methods, making glycan synthesis more efficient and precise.
Beyond that, we’ll explore some of the biggest challenges in the field, as well as the latest advancements that are helping to overcome them. And, of course, we’ll discuss the crucial role glycans play in biotechnology—especially in terms of biomolecule stability and their growing importance in therapeutic applications.
Introduction to Enzymatic Synthesis and Glycan Analysis
Your research focuses on enzymatic synthesis and glycan analysis. For those who may not be familiar with these concepts, could you explain what they entail and why they are so important in the field of biotechnology?
Of course! Enzymatic synthesis uses specialized enzymes to build or modify glycans—complex carbohydrate molecules—with high precision and efficiency. Enzymes like glycosyltransferases and glycosidases help assemble glycan structures with incredible specificity. This is crucial for developing glyco-conjugates used in vaccines, cancer therapies, biotechnology, and disease diagnostics.
Glycan analysis, on the other hand, studies the structure, composition, and function of glycans attached to proteins and lipids. Since glycans play key roles in biological processes, understanding them helps advance diagnostics, therapies, and vaccine development. Together, these fields drive innovation in personalized medicine and sustainable industrial processes.
Enzymatic synthesis has gained significant importance in recent years due to its specificity and efficiency. What are the main advantages of using enzymes for glycan synthesis compared to traditional chemical methods?
Unlike chemical synthesis, which often requires harsh conditions and can produce unwanted byproducts, enzymatic synthesis is highly specific and efficient. Enzymes act as biological catalysts, allowing reactions to occur under mild conditions while minimizing side reactions. This results in purer, more consistent glycans, making enzymatic synthesis a superior alternative for many applications.
Regarding glycan analysis, Asparia Glycomics utilizes advanced technologies for their characterization. What methodologies are employed in the laboratory to conduct this analysis, and what kind of information can be obtained from the samples?
Developing new methodologies for glycan synthesis and analysis is challenging due to the structural complexity and heterogeneity of glycans. Their highly branched and diverse nature makes both synthesis and characterization difficult, requiring precise enzymatic or chemical strategies. However, enzyme availability and specificity often limit synthesis efficiency, while scalable production remains a challenge.
On the analytical side, the lack of standardized databases and the complexity of biological samples make glycan identification and quantification difficult. Advanced techniques like mass spectrometry (MS) and liquid chromatography (LC) are essential but require specialized expertise and equipment, increasing costs and limiting accessibility. Additionally, integrating glycan analysis with other omics fields, such as genomics and proteomics, remains a major hurdle due to the absence of standardized cross-disciplinary tools.
Challenges and Methodological Development
Your research sits at the intersection of chemistry, biology, and biotechnology. What are the main challenges you encounter in developing new methodologies for glycan synthesis and analysis?
The biggest challenge in my opinion comes from the structural complexity of glycans—as previously said, they’re highly branched and diverse, making both synthesis and analysis difficult. On the synthetic side, enzymatic synthesis is limited by enzyme availability and specificity, making the scaling up process challenging.
Regarding the techniques, advanced techniques like MS and LC require specialized expertise and expensive equipment, which limits accessibility. Also, integrating glycan analysis with other omics fields, like genomics and proteomics, is another challenge due to the lack of standardized tools.
What are the most commonly used enzymes in enzymatic glycan synthesis, and what are their primary functions?
The most used enzymes in enzymatic glycan synthesis are glycosyltransferases (GTs). These enzymes are central as they transfer sugar units to acceptor molecules, building the glycan chains by forming specific glycosidic linkages. While GTs are the primary tools for constructing glycans, other types of enzymes also play essential roles in modifying and fine-tuning glycan structures.
For example, glycosidases are used to break or modify glycosidic bonds, removing or altering sugar residues. Epimerases change the stereochemistry of sugars, sulfotransferases add sulfate groups to glycans, and sialyltransferases transfer sialic acid to glycan chains. Though glycosyltransferases are the most widely employed, these other enzymes are equally important for adjusting the structure and function of glycans in a precise and controlled manner.
What are the current limitations of enzymatic glycan synthesis, and what strategies are being implemented to address them?
I´d say that enzymatic glycan synthesis faces several challenges, including limited enzyme specificity, scalability issues, glycan heterogeneity, and substrate limitations.
Strategies to overcome these challenges may include enzyme engineering to improve specificity, the development of multi-enzyme systems for more efficient synthesis, and advanced analytical techniques to monitor and control glycan structures. Additionally, efforts are focused on improving the availability and cost-effectiveness of sugar donors, standardizing synthesis protocols, and utilizing more biologically relevant environments, such as cell-free systems and microbial platforms, for production.
Structural and Functional Aspects of Glycans
How does the three-dimensional structure of a glycan influence its interaction with enzymes during synthesis?
It is a very interesting question: the 3D structure of a glycan determines how well it fits into an enzyme’s active site. Steric hindrance from bulky or branched glycans can slow down or block enzyme binding. Conversely, certain structural features can enhance enzyme recognition, ensuring efficient catalysis.
Is there a connection between enzymatic synthesis and glycoprotein bioengineering for therapeutic applications?
Enzymatic synthesis is essential for glycoprotein bioengineering because it allows precise control over glycosylation patterns, which directly impacts a protein’s stability, function, and immune response. This is crucial in developing biopharmaceuticals, vaccines, and targeted therapies. By fine-tuning glycan structures, we can optimize therapeutic proteins for better efficacy and aiming too for fewer side effects.
How does glycosylation impact the stability and function of biomolecules, and how can it be optimized through enzymatic synthesis?
As I just mentioned, glycosylation is critical for the stability, function, and activity of biomolecules, particularly proteins. The addition of specific sugar chains to proteins can enhance their stability by protecting them from degradation. It can also improve their solubility and influence their folding and conformation. But interestingly, glycosylation also affects protein interactions, recognition, and immune responses, which are crucial for therapeutic efficacy. Through enzymatic synthesis, glycosylation might be precisely controlled to optimize these properties or modifying sugar residues in a targeted manner, to ensure that the biomolecule of interest exhibits specific characteristics for its intended application, such as in biopharmaceuticals or vaccines.