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. 2020 Apr 20;10:78. doi: 10.1186/s13568-020-01012-8

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© The Author(s) 2020

Open AccessThis 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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Fig. 2 — Schematic representation of the plasmids used and generated in this study, and the resulting genetic modifications. a shows the vectors used in this study: pU-EV is the empty vector used as control, pU-XR is used for the overexpression of the xylose reductase gene XYL1, pU-XDH-KO for the knockout of the xylitol dehydrogenase gene XDH1, and pU-XDH-XR-KO for genomic replacement of XDH1 by XYL1. b Deletion of XDH1 by homologous recombination of the selection cassette (xdh∆::URA) and after release of URA5 by Ura-Blaster (xdhΔ) in the parental strain M. guilliermondii KU141F1. c Replacement of XDH1 by XYL1 (xdh∆::XR-URA) and after release of URA5 by Ura-Blaster (xdh∆::XR) in the parental strain KU141F1; AmpR: Ampicillin resistance gene; URA5: selection marker for uridine-prototrophy, P_ACT1: ACT1 promotor for constitutive expression of XYL1, T_TRP1: Terminator of XYL1 expression; REP: repetitive sequences for the use of the Ura-Blaster system to release URA5 selection marker; XDH1-upstr.: genomic sequence 1000 bp upstream of XDH1; XDH1-downst.: genomic sequence 1000 bp downstream of XDH1; S1, S2 and S3 indicate sequencing primer binding sites