We are pleased to announce the successful publication of a research article in the journal Microbial Cell Factories titled “Combining functional metagenomics and glycoanalytics to identify enzymes that facilitate structural characterization of sulfated N-glycans” as an initial outcome of an ongoing collaboration of glyXera with the Max Planck Institute for Dynamics of Complex Technical Systems (Magdeburg, Germany) and New England Biolabs (Ypswich, MA, USA) using our patented technology platform glyXboxCE, based on multiplexed capillary gel electrophoresis with laser-induced fluorescence detection (xCGE-LIF).
Although the biological importance of sulfated oligosaccharides has been widely recognized, there are only a few reports that describe sulfated N-glycans, which mostly occur in low abundance. This is largely due to a lack of dedicated analytical tools to detect and identify sulfated glycans and to elucidate their structures in detail, such as enzymes that enable verification of N-glycan sulfation. In a joint study, we combined functional metagenomic and glycomic screening to identify enzymes that specifically act upon sulfated N-glycans.
So far, screening for new enzymes is conducted on simple artificial sugar substrates to enable the required high throughput, but not on their natural counterparts. Our glyXboxCE technology allowed for screening directly on naturally occurring sulfated N-glycans and for optimization of the enzyme reaction conditions, both in high-throughput. The smart combination of high-performance methods on the metagenome, artificial sugar and naturally occurring N-glycan levels enabled a functional evaluation and characterization of glycoenzymes.
Our screen identified a sugar-specific sulfatase that allows for both selective enrichment and isolation of sulfated N-glycans, as well as specific removal of sulfate residues from N-glycans. Our joint study demonstrates not only the feasibility and power of combining functional metagenomic screening with high-performance glycoanalytics to discover enzymes that act upon chemical modifications of glycans, but also the absolute necessity to do so. The enzymes we discovered represent specificities that can help resolve the presence of sulfation in N-glycan structural analyses in combination with our highly sensitive glyXboxCE technology. Similar strategies could potentially be applied to other forms of post-glycosylation modifications.
The article is published open access and can be read in full on the publication web page: https://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-021-01652-w
If you would like to learn more about our xCGE-LIF-based glyXboxCE system and its wide range of possible applications, please feel free to get in touch.