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bioRxiv · 10.1101/2022.05.14.491371

NMR as a readout to monitor and restore the integrity of complex chemoenzymatic reactions

Marincin, K. A.·Hwang, Y.·Kengmana, E. S.·Meyers, D. J.·Frueh, D. P.
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Abstract

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 novel 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. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=78 SRC="FIGDIR/small/491371v1_ufig1.gif" ALT="Figure 1"> View larger version (15K): org.highwire.dtl.DTLVardef@11bc2e6org.highwire.dtl.DTLVardef@198ab6forg.highwire.dtl.DTLVardef@1f0747org.highwire.dtl.DTLVardef@116fcbf_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Marincin, K. A., Hwang, Y., Kengmana, E. S., Meyers, D. J., Frueh, D. P.. 2022-05-14. NMR as a readout to monitor and restore the integrity of complex chemoenzymatic reactions. https://doi.org/10.1101/2022.05.14.491371

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