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. 2021 Sep 28;12:5672. doi: 10.1038/s41467-021-25798-8

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© The Author(s) 2021

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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Fig. 2 — a Chemoenzymatic synthesis of 7 from 4 and a GnmT-SH and GnmP-coupled assay of GnmP-catalyzed S-methylation of 7 to 3 using 9 as a surrogate substrate. b HPLC analysis of metabolite profiles: (I) S. sp. CB01883 (wild-type), (II) SB21007 (ΔgnmP), and (III) SB21008 (ΔgnmP/gnmP). c HPLC analysis of in vitro assays: (I) substrate 9, (II) 9 + GnmT-SH (boiled) + 10, (III) 9 + GnmT-SH + 10, (IV) product 11, (V) 9 + GnmT-SH (boiled) + GnmP (boiled) + SAM, (VI) 9 + GnmT-SH + GnmP + SAM, and (VII) product 3. Substrate and enzyme concentrations used: 9, 0.5 mM; SAM, 2 mM; GnmT-SH, 10 μM; and GnmP, 30 μM. Incubation time: 20 min. The concentration of 10 used to trap 7 in situ as 11: 0.5 mM. “AU” denotes absorbance units.