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. 2021 Mar 5;11:5290. doi: 10.1038/s41598-021-84859-6

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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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Characterization of improved OsmY variants in 50-mL flask cultures. The OsmY-Tfu0937 activity, measured with the pNPG assay using an absorbance at 405 nm (left axis), are plotted as bars. The cell optical densities monitored at 600 nm (right axis) are shown as symbols. Cultures were inoculated using 1:100 dilution. (a) OsmY(WT) and variants were expressed using C41(DE3) in 2 × TY auto-induction media and at 37 °C and 200 rpm. The bars and symbols of the variants are represented using different colours; OsmY(WT) (black), OsmY(V43A) (blue), OsmY(S19C) (orange) and OsmY(TOA4) (red). Experiments were performed in triplicate. (b) OsmY(WT)-Tfu0937 and OsmY(TOA4)-Tfu0937 were expressed using C41(DE3) in 2 × TY auto-induction media, at 30 °C or 25 °C, and 200 rpm. The bars and symbols of the sample are represented using different colours; OsmY(WT) at 30 °C (black), OsmY(TOA4) at 30 °C (red), OsmY(WT) at 25 °C (grey), OsmY(TOA4) at 25 °C (magenta).