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LC-MS Ppant ejection assay of the in vitro cascade reaction of AzaBCMQ. EICs of Ppant fragments are shown. The HYAA-Ppant fragment (m/z331.1435) was detected in the presence of HAA, succinyl-CoA, ATP, FAD, and AzaBCMQ. The succinyl-HAA-Ppant fragment (m/z433.1751) and HYAA-Ppant fragment were observed in the no AzaB control reaction due to non-enzymatic production of4under these conditions (FigureS15). In the absence of AzaM, accumulation of the succinyl-HAA-Ppant fragment was detected. The HAA-Ppant fragment (m/z333.1591) was not detected. Three biological replicates were performed.

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