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. 2021 Aug 9;21:49. doi: 10.1186/s12896-021-00708-4

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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 — Correlation between recombinant protein titer and gene copy number. A The relationship between NK-Bs expression levels and aprN gene copy number were assessed using a plate assay. The copy number of the target gene in each strain is indicated below each colony. The circle around each colony represents the average halo size of responding multicopy expression strains. Scale bar, 1 cm. B SDS-PAGE analysis of NK-Bs secreted into the culture supernatant. M: protein molecular weight marker (the molecular weight of each band is indicated on the left); lanes 1–7: supernatants from shake-flask fermentation of NK-Bs 1–7 strains, respectively, collected after an 84-h induction with methanol. A total of 20 μL of each sample was loaded