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. 2020 Jan 28;10:1376. doi: 10.1038/s41598-020-57669-5

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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 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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Individual samples of ZmPSY1 protein purified from recombinant E. coli (Lot No. M20452-05) were heated for 15 minutes at a designated temperature ranging from 30–65 °C. Following this treatment, enzymatic production of 15-cis-phytoene was measured by HPLC. Panel A shows enzymatic activity (pmol μg-1 min-1) versus pre-incubation temperature, where the values are means + /− standard deviation of two technical replicates. Panel B shows HPLC chromatograms (287 nm) of chloroform:methanol extracts of selected ZmPSY1 activity assays. The area under the phytoene peak (retention time = 15.7 min) was used to calculate phytoene concentration. Pre-incubation temperatures are indicated to the right of each chromatogram trace (adapted from GR2E-FFP submitted study reports).