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. 2018 May 25;8:8121. doi: 10.1038/s41598-018-26445-x

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© The Author(s) 2018

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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Spectral changes during formation of AAO:p-anisic acid complex. (a) Native AAO, mixed with 0.5 mM of the ligand at 0.003, 0.01, 0.02, 0.03, 0.04, 0.05, 0.065, 0.085, 0.1, 0.13, 0.15, 0.2 and 0.4 s. (b) F397Y, mixed with 0.65 mM of p-anisic acid measured after 0.005, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3 and 4 s, and (c) F397W mixed with p-anisic acid 1 mM recorded at 0.005, 0.02, 0.04, 0.05, 0.07, 0.1, 0.2, 0.4, 0.5, 0.7, 1, 2 and 5 s. Insets show disappearance of initial species A and formation of final species B. The rate constant obtained from the global fitting of the spectral changes is shown. (d) Fits of the obtained k_obs for each variant and ligand concentration to equation (6). Native AAO (filled circles), and F397Y (open circles) and F397W (inverted triangles) variants.