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. 2021 Apr 6;11:7526. doi: 10.1038/s41598-021-86952-2

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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CD spectra and thermal denaturation curves. (A) CD spectra of WA20 and the N22A/H86K mutant of WA20 (ROWA) at 25 and 95 °C. (B) Thermal denaturation curves of WA20 and ROWA. The T_m values of the WA20 and ROWA proteins are 69.8 ± 0.4 and 80.4 ± 3.5 °C, respectively. (C) CD spectra of SUWA and the N22E/H86K mutant of SUWA (ROSA) at 30 and 150 °C. (D) Thermal denaturation curves of SUWA and ROSA. The T_m values of SUWA and ROSA are 124.7 ± 0.1 and 129.0 ± 0.1 °C, respectively. In the denaturation curves, the Y-axis represents the normalised value of [θ]_222 nm. The data of the denaturation curves were fitted to a two-state model with ΔC_p fixed to zero and the graphs were created using CDpal³³, version 2.18 (https://github.com/PINT-NMR/CDpal/).