GLYCONEER-An automated oligosaccharide synthesiser to transform glycobiology research within the University of York, and the UK glycoscience community

Lead Research Organisation: University of York
Department Name: Chemistry

Abstract

Macromolecules - literally big molecules - the proteins, DNA, RNA and carbohydrates - carry out the key roles in living organisms. Together they are responsible for all of the functions that sustain life, from metabolism through replication to the exchange of information between a cell and its environment. Carbohydrates, or sugars as they are sometimes known are the unsung heroes in all of this and the way they work, facilitating cell-cell communication and allow signalling throughout the body is not only of considerable scientific interest but is central in attaining a basic understanding of how an organism works or indeed fails to work, in the biotechnology industry through the use of sugars as additives in food processing and in development of new drugs in the pharmaceutical industry, all areas where the UK is a world leader.
Carbohydrates also constitute the largest source of biomass on Earth and their exploitation for novel applications in biomaterials, energy, food and health will be critical in moving away from dependence on hydrocarbons to develop sustainable biotechnologies and reduce GHG emissions, ensuring both energy and food security. Glycoscience is a broad term used for all research and technology involving carbohydrates, ranging from cell biology, human nutrition and medicine to carbohydrate-based materials and the conversion of carbohydrates to energy. The analysis, synthesis and biosynthesis of carbohydrates and their modification to industrial products are, therefore, central challenges in both industrial biotechnology and bioenergy. The last twenty years have seen a number of fundamental changes in the Glycosciences generating a technology push with respect to carbohydrate synthesis and modification, enzymology and glycomic analysis. At the same time, there is a technology pull - great demand and opportunities in diverse areas such as biopharmaceuticals (8 out of 10 top selling drugs worldwide are glycoproteins), foods (prebiotics designed for the human gut microbiota), antimicrobials (targeting cell surface recognition and biosynthesis), materials (from biorenewable polysaccharides) or energy (digesting the indigestible). However, research in this broad area is still fundamentally limited by the synthesis of complex carbohydrates, which can exist in multiple forms, and be linked together as 'polymers' in many ways with an almost unimaginable lack of limitations. While research into DNA and protein function has been stimulated by widespread access to synthesisers, machines that can literally be programmed by a user to produce any sequence of DNA or protein with almost no technical skills necessary, carbohydrates have been left behind because of their complexity. New technology advances mean that a commercially available carbohydrate synthesiser now exists with the capability to stimulate the glycoscience field to the same extent that synthesisers of DNA and proteins have in their own research fields in the last twenty years. With this new development, soon carbohydrates will no longer be known as the forgotten 'Cinderella' molecules of life.

Technical Summary

The last twenty years have seen a number of fundamental changes in the Glycosciences generating a technology push with respect to carbohydrate synthesis and modification, enzymology and glycomic analysis. However, research in this broad area is still fundamentally limited by the synthesis of complex carbohydrates, which can exist in multiple forms, and be linked together as polymeric oligo- and polysaccharides with little or no limitation to the spatial or stereo-arrangement. Coupled with this inherent complexity, is the lability of the monosaccharide building blocks themselves, and the depth of literature regarding oligosaccharide synthesis built up over the last fifty years, and the necessity for endless rounds of purification. While research into DNA and protein function has been stimulated by widespread access to synthesisers, which can be pre-programmed to produce bespoke oligonucleotides or proteins of any almost any sequence, and little restriction in length, using commercially available building blocks. Carbohydrates have been left behind because of their complexity, and the lack of an automated method for their synthesis. Purchase of UK's first fully automated oligosaccharide synthesiser- the 'Glyconeer', will transform UK glycoscience by erasing the boundaries between traditional synthetic glycochemistry and glycobiology and rewriting the "rules" of interdisciplinary glycoscience research. Access to the 'Glyconeer' will confer any researcher, irrespective of their primary discipline, with the capability to produce pure, bespoke glycopolymers for wide ranging research applications. Our vision is that the 'Glyconeer' will provide a stimulus to the glycoscience research effort, equivalent to the effect that widespread use of oligonucleotide and peptide synthesisers have had on the fields of glycomics and proteomics. With this new development, soon carbohydrates will no longer be known as the forgotten 'Cinderella' molecules of life.

Planned Impact

Who will benefit from this research and how?
Locating an automated oligosaccharide synthesiser in York, operated on the UK-wide accessible basis outlined in this proposal will be of immediate and direct benefit to the academic and commercial National carbohydrate community, as reflected in the broad institutional base of the Investigators and the Expression of Interest letters. *A key goal of this work is to impact biomedical research (fertility and antibacterial focus) through development of new diagnostic tests and treatments to be delivered by healthcare professionals, yielding ultimate improvements to the Nation's health with concurrent impact on healthcare policy.*We will also deliver important developments in bioenergy and therefore impact the Biotechnology sector, particularly Industrial companies developing enzymatic and microbial toolkits. On a broader level, finding bio-tools to unlock the potential of plants to act as solar fuel factories will alter the economic viability of developing renewable fuels which will make non fossil fuel technology an affordable reality, ultimately requiring energy sector changes on a policy and engineering level. *Proof of the transformative power of automated synthesis in uniting chemical and biological research development will also further the growth of novel Chemical Engineering approaches which again resonates across the academic and industrial sectors and is anticipated to be a wealth generating activity. *Educating the Public, particularly school children, about the important applications of understanding bioscience and developing new methodological toolkits will inspire a future generation of scientific leaders. *We will have impact on Museum research and our fundamental understanding of human development via the delivery of new tests for monitoring carbohydrates in the environment, permitting the use of complex sugars as novel chemical signature for dating ancient human artefacts. This is an important cultural output. *Finally, all our work will continue to raise the profile of the UK as a world-leading scientific nation which supports and embraces cutting-edge approaches to tackle challenging problems of key importance. This is of vital importance in maintaining an ambitious, problem solving National cultural identity.

Publications

10 25 50
 
Description The Glyconeer has been established as a platform technology in the UK for synthesising complex oligosaachrides, including compounds which have potential biotechnological/biomedical applications in the study of neurodegenerative disease, and bacterial infections. Collaborations with users of the Glyconeer have been established both in the UK and internationally.
Exploitation Route Synthesis of complex oligosaccharides and peptides for biotechnological/ biomedical applications
Sectors Agriculture, Food and Drink,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description On-going. It is anticipated that ongoing collaborations with academic partners in the UK and in New Zealand, may potentially result in translational impact in the synthesis of pharmaceutically relevant oligosaccharides. A PCT application has been filed by the UoY which utilises peptides accessible through the Glyconeer.
First Year Of Impact 2016
Sector Agriculture, Food and Drink,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
 
Description Automated synthesis of heparan sulfate oligosaccharides, Royal Society International exchange
Amount £12,000 (GBP)
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom
Start 04/2016 
End 04/2018
 
Description BBSRC IBCarb NIBB Proof of Concept funds
Amount £100,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 02/2016 
End 11/2016
 
Description Investigation of a Novel Bacterial Enzyme Involved in Cardiovascular Device Infections
Amount £242,000 (GBP)
Funding ID PG/16/5/31912 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2016 
End 07/2019
 
Description International exchange 
Organisation Victoria University of Wellington
Department Ferrier Research Institute
Country New Zealand 
Sector Academic/University 
PI Contribution Royal society international exchange visits between the Fascione lab and the Ferrier Institute in New Zealand, visits to Ferrier institute by Dr Fascione and PDRA to develop skills in the synthesis of complex disaccharides.
Collaborator Contribution Visit to Fascione lab to train on the Glyconeer by Dr Ralf Schwoerer and Prof Peter Tyler. Supply of complex disaccharides for automated assembly of heparin sulfate glycans.
Impact None as yet
Start Year 2016
 
Description Research Collaboration 
Organisation University of York
Department Department of Chemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution Research collaboration on the automated synthesis of macrocycles between Dr Fascione and Dr Will Unsworth. Training of PhD students on the Glyconeer
Collaborator Contribution Synthesis and supply of macrocycle precursor building blocks
Impact None as yet
Start Year 2016
 
Description Research Collaboration 
Organisation University of York
Country United Kingdom 
Sector Academic/University 
PI Contribution Research collaboration between PI and Dr Fascione with Prof Jen Potts, Prof Gavin Thomas, focussed on exploration of glycan biosynthesis. Supply of oligosaccharides synthesised on Glyconeer.
Collaborator Contribution Supply of enzymes and proteins for characterisation of oligosaccharide substrates
Impact Multidisciplinary grant funding
Start Year 2015
 
Description Research collaboration 
Organisation Keele University
Country United Kingdom 
Sector Academic/University 
PI Contribution Collaboration between Dr Gavin Miller and Dr Martin Fascione on the synthesis of glycomimetic compounds, associated with Dr Miller's EPSRC funded research
Collaborator Contribution Training of Miller group PhD on the Glyconeer, to enable synthesis go glycomimetic compounds
Impact None as yet
Start Year 2017