Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry. This journal is © The Royal Society of Chemistry 2020 Supporting Information Modular Control of L-Tryptophan Isotopic Substitution via an Efficient Biosynthetic Cascade Thompson, C. M.; McDonald, A. D.; Yang, H.; Cavagnero, S.*; Buller, A. R.* Department of Chemistry, University of Wisconsin-Madison 1101 University Avenue Madison, WI 53706 *corresponding authors: Andrew R. Buller, email:
[email protected]; Silvia Cavagnero, email:
[email protected] Table of contents 1. General information 2 2. Experimental section 2 2.1 Materials 2 2.2 Methods 2 2.2.1 Cloning, expression, and purification of PfTrpB2B9 and TmLTA 2 2.2.2 Optimization of TmLTA-TrpB2B9 cascade 3 2.2.3 Preparative-scale synthesis of Trp isotopologs 4 2.2.4 Analytical- and preparative-scale chromatography 5 2.2.5 Determination of enantiopurity of Trp isotopologs 5 2.2.6 NMR analysis 6 3. Characterization of Trp and Trp isotopologs 7 4. Supporting references 10 5. Supporting tables and figures 11 - 17 − 1 − 1. General information The glassware used in the reactions carried out in this work was thoroughly washed, and all experiments were executed following necessary safety precautions. Evaporation of solvents was performed at reduced pressure using a rotary evaporator. Electronic-absorption measurements were done with a UV-2600 spectrophotometer (Shimadzu). 2. Experimental section 2.1. Materials Chemicals and solvents were obtained from commercial suppliers and used without further 13 purification: formaldehyde-D2 (Cambridge Isotope Laboratories, Inc.); (2- C)glycine (Cambridge Isotope Laboratories, Inc.); D2O 99.9% (Sigma-Aldrich); indole (Sigma- Aldrich); pyridoxal 5’ monophosphate; (Sigma-Aldrich) L-serine (Sigma-Aldrich).