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. 2020 Feb 14;11:899. doi: 10.1038/s41467-020-14754-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Structural comparison of EndoBT-3987_D312A/E314L-Man₉GlcNAc₂Asn in gray and EndoH (PDB CODE 2EBN) in blue. The residues involved in substrate recognition are colored by loops: loop 1 (yellow), loop 2 (orange), and loop 3 (pinked). b Structural comparison of the HM-type glycan conformation in the active site of EndoBT-3987_WT-Man₉GlcNAc-2 (gray) and EndoS2 (PDB CODE 6MDV) (green) on surface representation of binding site of EndoS2 (yellow). c Structure weighted sequence alignment of BT3987 with GH18 ENGases family with characterized endo-N-acetyl-β-D-glucosaminidase activity against HM-type N-glycans and inactive against CT-type N-glycans. Comparison of BT3987 from B. thetaiotaomicron VPI-5482 (Q8A0N4, Uniprot code), EndoH from Streptomyces plicatus (P04067, Uniprot code), EndoF1 from Elizabethkingia meningoseptica (P36911, Uniprot code), A6286 from Prevotella melaninogenica (D9RSV7, Uniprot code), BT1285 from B. thetaiotaomicron VPI-5482 (Q8A889, Uniprot code), EF2863 from Enterococcus faecalis (Q830C5, Uniprot code), Endo-Fsp from Flavobacterium sp. (P80036, Uniprot code), Eng18B from Hypocrea atroviride IMI 206040 (G9P8KO, Uniprot code), Eng18A from Hypocrea atroviride IMI 206040 (G9NR36, Uniprot code), EndoT from Hypocrea jecorina (C4RA89, Uniprot code), and EndoFv from Flammulina velutipes (D1GA49, Uniprot code). The catalytic residues are marked with red dots and the key residues that interact with Man₉ are marked with green dots.