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. 2021 Aug 26;12:5139. doi: 10.1038/s41467-021-25233-y

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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 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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Fig. 2 — a The concentration of rewired BDO cell extract was adjusted to maximize conversion of acetoin to BDO while minimizing ethanol production. b pH was varied in physiologically compatible ranges that would not be possible in vivo to highlight the disparate pH optima for BDO and ethanol biosynthesis in vitro. c NAD and ATP concentrations were covaried to optimize the cofactor pool. d Glucose concentration was varied from 20–200 mM to compare the ratio of BDO to ethanol and the final BDO titer. Data represent mean ± standard deviation of n = 3 technical replicates. Source data are provided as a Source Data file.