biostudies-literatureMarincin KAFlight Attendant Medical Research InstituteNCATS NIH HHSInstitute for Clinical and Translational Research, University of Maryland, BaltimoreNational Institutes of HealthNational Institute of General Medical SciencesNIGMS NIH HHS107265https://www.ebi.ac.uk/biostudies/studies/S-EPMC9463103biostudies-literatureUnknown342The non-invasive nature of NMR offers a means to monitor biochemical reactions in situ at the atomic level. We harness this advantage to monitor a complex chemoenzymatic reaction that sequentially modifies reagents and loads the product on a nonribosomal peptide synthetase carrier protein. We present a protocol including a pulse sequence that permits to assess both the integrity of reagents and the completion of each step in the reaction, thus alleviating otherwise time-consuming and costly approaches to debug and repeat inefficient reactions. This study highlights the importance of NMR as a tool to establish reliable and reproducible experimental conditions in biochemical studies.Journal of magnetic resonance (San Diego, Calif. : 1997)NMR as a readout to monitor and restore the integrity of complex chemoenzymatic reactions.PMC9463103R01 GM104257UL1 TR003098R01GM104257UL1TR003098T32 GM080189Meyers DJKengmana ESMarincin KAFrueh DPHwang YfalseNMR as a readout to monitor and restore the integrity of complex chemoenzymatic reactions.The non-invasive nature of NMR offers a means to monitor biochemical reactions in situ at the atomic level. We harness this advantage to monitor a complex chemoenzymatic reaction that sequentially modifies reagents and loads the product on a nonribosomal peptide synthetase carrier protein. We present a protocol including a pulse sequence that permits to assess both the integrity of reagents and the completion of each step in the reaction, thus alleviating otherwise time-consuming and costly approaches to debug and repeat inefficient reactions. This study highlights the importance of NMR as a tool to establish reliable and reproducible experimental conditions in biochemical studies.2022-01-01T00:00:00Z2022 Sep2026-05-28T01:36:13.567Z2024-11-09T11:20:10.345ZS-EPMC94631033584997310.1016/j.jmr.2022.107265