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. 2020 Jan 31;10:1589. doi: 10.1038/s41598-020-58559-6

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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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(a) Scheme of the mechanism of hydrolysis of substrate 1 catalyzed by MuV-HN neuraminidase. The 3D Symbol Nomenclature For Glycans (SNFG) has been used. Chem Draw 2006 Software (http://www.cambridgesoft.com) was used to draw the sugar moieties. (b) Section of ¹H-NMR spectra at different time of the enzymatic reaction have been reported. 7.5 μM of MuV-HN and 550 μM of 1 were solved in PBS deuterate buffer (pH = 7). The NMR quantification of the substrate concentration was performed by integration of the well-dispersed resonances of 1 (H1 Gal, H3_eq Neu5Ac). The NMR spectra were plotted by using the xwinplot tool of TopSpin 3.6.1 software (www.bruker.com). (c) Analysis of the MuV-HN kinetics toward substrate 1 by means of the explicit reformulation of the integrated form of MM equation with Lambert W fit as solution. The plot of the concentration of 1 as function of time has been reported. The substrate concentration was evaluated from the H3_eq resonance of Sia unit. The fit of the kinetic data by the Lambert-W fit afforded a K_M value of 12 μM and V_max of 7 * 10⁻³ (mM/min). The blue dashed line represented the confidence interval of the fit. Data were analyzed by Sigma Plot 11.0 (http://www.sigmaplot.co.uk/).