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. 2021 Jan 7;20:9. doi: 10.1186/s12934-020-01502-1

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© The Author(s) 2021

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Expression of 6HFh8-GFs in flask culture. The proteins were induced by lactose in auto-induction media. They were then purified by HisTrap chromatography. a SDS-PAGE analysis of protein expression. Red dot, the fusion protein. I: insoluble fraction; S: soluble faction; T: total fraction. The images are representative of two independent experiments. b Quantitative analysis of protein expression. The bar graphs show percent expression (black bar) and solubility (gray bar) (primary y-axis), and the red line indicates expression level times solubility (secondary y-axis). Red line peak indicates the optimal temperature for fusion protein expression. Except for SCF and TIMP-1, all proteins were successfully expressed in soluble form at low temperature (25 °C and/or 30 °C). The expression level and solubility were analyzed by densitometry using ImageJ with two independent experiments