Glycoenzymes for Bioindustries
Lead Research Organisation:
Newcastle University
Department Name: Biosciences Institute
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
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.
Publications
Briliute J
(2019)
Complex N-glycan breakdown by gut Bacteroides involves an extensive enzymatic apparatus encoded by multiple co-regulated genetic loci.
in Nature microbiology
Briliute Justina
(2016)
Insight into N-glycan breakdown by the gut microbiota
in GLYCOBIOLOGY
Crouch LI
(2022)
Plant N-glycan breakdown by human gut Bacteroides.
in Proceedings of the National Academy of Sciences of the United States of America
Crouch LI
(2020)
Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown.
in Nature communications
Crouch Lucy I.
(2016)
Degradation of complex N-glycans by gut Bacteroides species
in GLYCOBIOLOGY
Cuskin Fiona
(2016)
Pivotal alpha mannosidase generates specificity for N-glycans through requirement for GlcNac at+2 subsite.
in GLYCOBIOLOGY
Guo BS
(2018)
Cloning, purification and biochemical characterisation of a GH35 beta-1,3/beta-1,6-galactosidase from the mucin-degrading gut bacterium Akkermansia muciniphila.
in Glycoconjugate journal
Rebello OD
(2020)
A novel glycosidase plate-based assay for the quantification of galactosylation and sialylation on human IgG.
in Glycoconjugate journal
| Description | Discovered and characterised 10s of novel O- and N-glycan active glycosidases that have potential uses in glycoprofiling and glycan analysis. Several of these enzymes have been licenced for use by Ludger who will use them in-house, but also sell to interested customers. One enzyme was considered to have such broad commercial and academic potential it has been patented by Newcastle and Ludger. |
| Exploitation Route | Use of well characterised and commercially available glycoenzymes in medicine, industry and academia. |
| Sectors | Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology |
| Title | Glycosidases licenced to Ludger |
| Description | 7 glycosidases targeting different N- and O-glycan structures discovered and characterised by Newcastle have been licenced for use and sale by Ludger Ltd. |
| IP Reference | |
| Protection | Protection not required |
| Year Protection Granted | 2019 |
| Licensed | Commercial In Confidence |
| Impact | None yet. Licencing only recently completed. |
| Title | PNGaseL |
| Description | N-glycosylation is a common form of post translational protein modification. It results in the formation of a glycoprotein, in which an N-glycan is attached to an asparagine residue of the protein. Glycans have a wide variety of functions, including protecting the protein from proteases and binding interactions. Given that glycans can modulate protein function, removal of N-glycans from proteins may be desirable in certain circumstances. All N-glycans have a core structure, but some details of their decoration vary according to the organism that synthesised them. For instance, glycosylated proteins from mammalian, insect and plant cells vary in their N-glycan profile. Mammalian-derived glycosylated proteins comprise a-1,6 core fucosylated N-glycans; insect proteins comprise a mixture of a-1,6 and a-1,3 core fucosylated N-glycans; and plant proteins comprise a-1,3 core fucosylated N-glycans only. Mammalian proteins also contain glycans with sialic acid caps which can vary, and hydrolyse, under acid conditions. The Peptide-N4-(N-acetyl-ß-glucosaminyl)asparagine amidase (PNGase) enzyme family hydrolyses the ß-aspartylglucosaminyl linkage between an N-glycan and the asparagine residue to remove the glycan from the protein. These enzymes are produced naturally by many organisms for different functions, for example, to acquire N-glycans for nutrients or to recycle or remodel glycoproteins. A number of PNGase enzymes have been previously characterised, the majority of which only remove a subset of N-glycans. For example, PNGase F from Elizabethkingia menigoseptica is capable of removing a-1,6 core fucosylated N-glycans only (e.g. from glycosylated proteins expressed in mammalian systems). It is the most common PNGase that is used by the scientific community. By contrast, PNGase F type II (also from Elizabethkingia menigoseptica) and PNGase A are specific to removal of a -1,3 core fucosylated N-glycans (such as those found on glycosylated proteins expressed by plant cells). PNGase A was originally characterised from almonds, but homologues of this enzyme are now also sold commercially. Glycoproteins must be pre-treated with trypsin before PNGase A can effectively be used. Of the characterised PNGases, only PNGase H+ from Terriglobus roseus is capable of removing both a-1,6 core fucosylated N-glycans and a-1,3 core fucosylated N-glycans. However, the low pH conditions required by PNGase H+ may adversely affect the glycoprotein and/or N-glycan products of the reaction, which restricts its use. Acid hydrolysis of sialic acid caps, as well as protein denaturation are particular problems associated with PNGases that have a low optimum pH. There is a need for an improved means for removing N-glycans from glycoproteins. |
| IP Reference | PCT/GB2021/052550 |
| Protection | Patent application published |
| Year Protection Granted | 2021 |
| Licensed | Commercial In Confidence |
| Impact | None so far, product is still being developed for commercial release. |