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. 2017 Jun 5;7:2768. doi: 10.1038/s41598-017-02382-z

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© The Author(s) 2017

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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Binding assays of BP-ChBD_CHIT1-49 towards different ligands. (A) Insoluble chitin was first used as a ligand to compare the binding efficiency of the protein used in the present study (BP-ChBD_CHIT1-49) and the one reported in our previous work²¹ (BP-ChBD_CHIT1-72) using the reporter enzymatic activity of the carrier protein BlaP. The β-lactamase BlaP without any inserted ChBD_CHIT1 (labelled BP) was used as a negative control (B) Hyaluronan (HA; 50% acetylated; black) and peptidoglycan (PG; 50% acetylated; hatched) were also tested using the same procedure. (C) GlcNAc₁, GlcNAc₂, GlcNAc₄ and GlcNAc₆ (respectively G₁, G₂, G₄ and G₆) were used as competitors. An equimolar amount of competitor was pre-incubated with the protein before incubating the mixture with insoluble chitin. Except for GlcNAc₁ (*), all competitors showed a similar inhibition effect (**).