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. 2022 Nov 7;14(1):2132903. doi: 10.1080/19490976.2022.2132903

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© 2022 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Recombinant BaiA1 from L. scindens ATCC 35704 catalyzes the final reductive step in the formation of allo-DCA and allo-LCA. (a) Formation of bile acid stereoisomers after reduction of 3-oxo-allo-LCA and 3-oxo-allo-DCA by 3α-HSDH and gene organization of baiA1 in L. scindens ATCC 35704. (b) Cloning strategy of baiA1 and baiA1 + baiP in pETduet. SDS-PAGE of His-tagged purified recombinant BaiA1 and BaiA1 + BaiP expressed in E. coli BL21(DE3). (c) Representative LC/MS chromatograms after resting cell assay with E. coli BL21(DE3) pETduet_Control or pETduet_BaiA1 incubated in anaerobic PBS containing 50 μM 3-oxo-allo-LCA (Top panels 1 & 2), E. coli BL21(DE3) pETduet_BaiP-BaiA1 incubated with 50 μM 3-oxo-Δ⁴-LCA (Panel 3) and E. coli BL21(DE3) pETduet_BaiP-BaiA1 incubated with 50 μM 3-oxo-Δ⁴-DCA (Panel 4). Standards are shown in Panel 5 (bottom). The overall two-step reaction is shown on the panels.