Glycoenzymes for Bioindustries

Lead Research Organisation: Institute of Food Research
Department Name: Gut Microbes and Health

Abstract

There are a wealth of glycoenzymes in nature but only a small number of enzymatic activities are used in industrial processes, primarily carbohydrate degrading enzymes used in high volume manufacturing such as amylases/xylanases/cellulases/oxidases used in baking/detergents/biofuels. Even fewer examples exist in high value manufacturing applications such as biologics: one example is the production of the therapeutic Cerezyme where alpha-neauramindase is used to deglycosylate exposing terminal Man residues improving its targeting. A major hurdle is the availability of appropriate enzymes which can limit utilisation in the early stage of process development thus reducing the number of successful processes involving glycoenzymes. The present project aims to overcome the current limitations of glycoenzyme availability by bringing together a number of innovative high-throughput approaches to biocatalyst discovery. We have assembled a list of 20 different glycoenzyme activities including various glycosyltransferases, polysaccharide-modifying enzymes, glycosidases and sugar oxidases that will form the basis of the research. Through iterative rounds of biocatalyst discovery, characterisation and development we aim to bring to market panels of different glycoenzyme classes that are readily available for screening by end users.

Technical Summary

The project will translate existing academic work on the development of novel glycoenzymes to produce a suite of stable/robust enzymes at scale to address the current gaps in certain commercially available specificities. The lack of availability of such enzymes is a major barrier to those wishing to utilise them for diagnostics, synthesis or chemical modification processes often necessitating de novo synthesis of the enzyme targeting the required reaction before development/evaluation of the process can begin. Each academic partner will develop a subset of the glycoenzyme classes, providing the necessary analytical expertise to characterise and optimise the enzymes. Underpinning this work will be genome mining combined with enzyme evolution to discover/develop novel microbial sourced enzymes displaying desired activities. Industry input to the project will guide the development of the enzymes whilst ensuring that they are fit for purpose and brought to market to allow end user access.

Planned Impact

As described in proposal submitted to IUK

Publications

10 25 50
 
Description We have identified and characterised novel glycoenzymes of interest for a range of industrial applications.
Exploitation Route The glycoenzymes could be used as tools for glycan synthesis
The glycoenzymes could be used as tools for glycan profiling
The glycoenzymes could be used as tools for diagnosis
Sectors Agriculture, Food and Drink,Pharmaceuticals and Medical Biotechnology

 
Description The glycoenymes identified in this project are subject of MTA with industry.
Sector Agriculture, Food and Drink,Pharmaceuticals and Medical Biotechnology
 
Description Carbohydrate analysis 
Organisation John Innes Centre
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution Supervised and coordinated research project
Collaborator Contribution Provided expertise and access to NMR facility
Impact WO2017134466A1. Patent Application on intramolecular trans-sialidase; Carbohydr Res. 2017 Nov 8;451:110-117. doi: 10.1016/j.carres.2017.08.008.
Start Year 2007
 
Description Fucosidase testing 
Organisation BASF
Country Germany 
Sector Private 
PI Contribution We have produced and characterised novel fucosidases as part of the Glycoenzyme project
Collaborator Contribution A subset of fucosidases were provided to BASF under MTA for them to test suitability in potential applications
Impact Transfer of materials
Start Year 2017
 
Description Fucosidase testing 
Organisation Ludger Ltd
Country United Kingdom 
Sector Private 
PI Contribution We have produced and characterized novel fucosidases as part of the Glycoenzyme project
Collaborator Contribution A subset of fucosiadses have been tested by Ludger for potential application
Impact Exchange of materials, Technical reports
Start Year 2016
 
Description STD-NMR 
Organisation University of East Anglia
Department Schools of Pharmacy and Medicine UEA
Country United Kingdom 
Sector Academic/University 
PI Contribution We provided proteins and ligands for analysis by STD NMR to complement some of our data on mechanisms of host-bacteria interactions. This collaboration resulted in high impact joint publications led by QIB (Proc Natl Acad Sci U S A. 2018 pii: 201715016; Nat Commun. 2017 ;8(1):2196)
Collaborator Contribution STD- NMR analysis of proteins and ligands we provided. Some of this work led to the development of DEEP STD NMR, leading to a joint publication led by UEA (Angew Chem Int Ed Engl. 2017 56:15289-15293).
Impact Proc Natl Acad Sci U S A. 2018 pii: 201715016; Nat Commun. 2017 ;8(1):2196; Angew Chem Int Ed Engl. 2017 56:15289-15293
Start Year 2014
 
Description Synchrotron Oxford 
Organisation Diamond Light Source
Country United Kingdom 
Sector Academic/University 
PI Contribution Provided collaborator with proteins and ligands for X-ray crystallography and cryo-EM
Collaborator Contribution X-ray crystallography of proteins free and in complex
Impact Nat Commun. 2017 Dec 19;8(1):2196. doi: 10.1038/s41467-017-02109-8.
Start Year 2016
 
Description Glycobiology Network Latin America 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Workshop on Glycobiology of host-microbe interactions in the gut. Biotechnology and glycobiology tools for human health, Guanajuato, Mexico, 9-11/10/2017
Year(s) Of Engagement Activity 2017