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ToolsBakshani, Cassie R., Urbanowicz, Paulina A., Tortosa, Conchi Badia, Melo Diaz, Javier M., Kujawska, Magdalena, Ojuri, Taiwo O., Hall, Lindsay J., Spencer, Daniel I. R., Bolam, David N. and Crouch, Lucy I. (2025) PNGaseL from Flavobacterium akiainvivens targets a diverse range of N-glycan structures. Royal Society Open Science, 12 (8). ISSN 2054-5703
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Abstract
Peptide:N-glycosidases (PNGases) are used by a wide range of organisms to remove N-glycan structures from proteins for use as either nutrients or in glycoprotein processing. PNGaseF is the most well-characterized enzyme of this family and is widely used in glycobiology to allow study of the N-glycome of a specific protein, cell and tissues, for instance. Despite this, PNGaseF has limitations in the types of N-glycan structures it can target. In this study, we explored the specificities of six uncharacterized PNGases selected from diverse parts of the PNGaseF superfamily. One of these enzymes, PNGaseL from Flavobacterium akiainvivens, is the highlight of this study due to its very broad specificity, exemplified by its ability to cleave mammalian-, plant- and invertebrate-type complex N-glycans as well as high-mannose N-glycans. A detailed biochemical and structural characterization was carried out against a variety of substrates to illustrate the advanced capability of PNGaseL in comparison to the canonical PNGaseF and PNGaseA enzymes. To determine the optimal reaction conditions, assess stability and define limitations of PNGaseL, a series of validation studies were performed. The data reveal that PNGaseL has potential utility in a range of glycobiology applications that are superior to the current commercially available options.
| Item Type: | Article |
|---|---|
| Additional Information: | Data accessibility: Supplementary material is available online [Bakshani CR et al.. 2025 Supplementary material from: PNGaseL from Flavobacterium akiainvivens targets a diverse range of N-glycan structures. Figshare. (doi:10.6084/m9.figshare.c.7979929)]. Funding information: The work was funded by The Academy of Medical Sciences (SBF0061175), the Wellcome Trust and Royal Society Sir Henry Dale fellowship (224240/Z/21/Z) awarded to L.I.C. and a BBSRC/Innovate UK IB catalyst award to D.N.B. 'Glycoenzymes for Bioindustries' (BB/M029018/1). T.O.O. is funded by the BBSRC Midlands Integrative Biosciences Training Partnership (MIBTP) with his studentship in collaboration with industrial partners Ludger (Oxford, UK) awarded to L.I.C. Acknowledgements |
| Uncontrolled Keywords: | chromatography,crystal structures,glycobiology,modelling,n-glycans,nutrient acquisition,pngase |
| Faculty \ School: | Faculty of Medicine and Health Sciences > Norwich Medical School |
| UEA Research Groups: | Faculty of Medicine and Health Sciences > Research Centres > Metabolic Health |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 18 Sep 2025 08:31 |
| Last Modified: | 22 Sep 2025 09:30 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/100431 |
| DOI: | 10.1098/rsos.251012 |
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