Unity inova 500 mhz instrument

All solvents, chemicals, and reagents were purchased from Millipore Sigma, unless otherwise stated. AMS was purchased from Synnovator, Inc. Samples for LC-MS were prepared in 0.45 μM polytetrafluoroethylene mini-UniPrep vials from Agilent. All prep-HPLC was performed using an Agilent/HP 1050 quaternary pump module with an Agilent/HP 1050 MWD module with a Phenomenex Luna 10u C18(2) 100A column, 250 × 21.20 mm, 10 μm with guard column. All LC-MS was performed on an Agilent 6130 quadrupole LC-MS with G1313 autosampler or G1367B autosampler, G1315 diode array detector, and 1200 series solvent module. A Phenomenex Gemini C18 column, 50 × 2 mm, 5 μm with guard column was used for all LC-MS separations. Mobile phases for prep-HPLC and LC-MS were 0.1% formic acid in (A) H₂O and (B) CH₃CN, and data were processed using ChemStation software (Agilent). NMR was performed on a Varian Unity Inova-500 MHz instrument. The maximum likelihood phylogenetic trees of selected NIS synthetases were built by the “one-click” method from NGPhylogeny.fr (56 (link)) and visualized using iTOL v6 (57 (link)). Sequence alignments of NIS synthetases were performed by the Clustal Omega online tool from EMBL (58 (link)). The alignment results were viewed in ESPript 3.0 (59 (link)) and the secondary structure was denoted based on HSC-AMS–bound SgDesD structure (PDB: 7TGM).