Fig. 6: LC-MS analysis of in vitro assays with FosH and FosM. | Nature Communications

Fig. 6: LC-MS analysis of in vitro assays with FosH and FosM.

From: Covalent warhead assembly in fostriecin biosynthesis involves malonylation-lactonisation by a bifunctional thioesterase and enzymatic demalonylation

Fig. 6: LC-MS analysis of in vitro assays with FosH and FosM.

a EIC of FosM reaction with malonyllactone 27, and the no-enzyme control reaction. b Simultaneous incubation of FosH and FosM with 27, and the negative control without FosM. c Reaction of FosM with 33 and the negative control without enzyme. d FosH reaction with AUDL 28 and negative control without enzyme. e FosMod8 and FosTEII were incubated with substrate 17, malonyl-CoA, and NADPH for 3 h. Subsequently, FosH, ATP, and MgCl2 were added to the reaction, and the mixture was incubated for an additional hour (top). Full reconstitution: FosMod8 and FosTEII were incubated with substrate 17, malonyl-CoA, and NADPH for 3 h. Subsequently, FosH, FosM, ATP, and MgCl2 were added to the reaction, and the mixture was incubated for an additional hour (bottom). Dashed arrows illustrate the emergence of side products 37 and 24 during the reactions. Supplementary Figs. 36–39 for HRMS(/MS) data.

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