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. 2021 Aug 16;20:158. doi: 10.1186/s12934-021-01653-9

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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. 4 — Characterization of kstd-knockout and ksh-knockout mutants for phytosterol fermentation in small-scale fermentation. a TLC assay of the catabolites by kstd-knockout and ksh-knockout mutants during phytosterol fermentation, compared with those of the WT strain and the standard samples of 4-AD, ADD and 9OH-AD. ΔKstD1, ΔKshB122, ΔKshA226 and ΔKshA395 refer to the HGMS2^Δkstd211, HGMS2^{Δkstd211 + ΔkshB122}, HGMS2^{Δkstd211 + ΔkshA226} and HGMS2^{Δkstd211 + ΔkshA395} mutants, respectively. Weak 9OH-AD spots for the samples from the HGMS2^Δkstd211, and HGMS2^{Δkstd211 + ΔkshA226} mutants are marked by arrows. b HPLC profile of the extracts from the phytosterol fermentation by three double mutants for 144 h compared with those of the WT strain and the HGMS2^Δkstd211 mutant. (1) 4-AD; (2) ADD; (3) 9OH-AD and (4) BA. Asterisk: unknown HGMS2 metabolites when the strain was cultured without phytosterol