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. 2020 Sep 23;10:15493. doi: 10.1038/s41598-020-72418-4

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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 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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Multiple sequence alignment of ancIPMDH-IQ, ancIPMDH-ML, T. thermophilus IPMDH, B. subtilis IPMDH, and S. cerevisiae IPMDH. Numbers above the sequences are those of the ancestral enzymes. Residues conserved among the five sequences are highlighted in green. V277 and H278, the two residues mutated in the ancestral sequences, producing ancIPMDH-IQ-VAHG and ancIPMDH-ML-VAHG, are boxed.