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Figure S1. Comparison of LC-ESI-Orbitrap-MS profiling of metabolites present in the crude supernatant. (A) Total ion chromatogram (TIC) of P. larvae DSM25430 wt and (B) total ion chromatogram (TIC) of P. larvae DSM25430 ∆pamA. Paenilamicin could not be detected directly from crude supernatants due to early elution in the dead volume and ion suppression. The secondary metabolite sevadicin is indicated with a red arrow.

Figure S2. MS/MS fragmentation studies on paenilamicin by HPLC-ESI-triple quadrupole MS from P. larvae secretome after sample workup. (A) MS/MS fragmentation spectrum with [M + 2H]2+ = 505.4 as precursor ion (♦). (B) MS/MS fragmentation spectrum with [M + 2H]2+ = 519.4 as precursor ion (♦).

Figure S3. HPLC UV analytics of pure paenilamicin. (A) Chromatogram of pure paenilamicin monitored at λ = 214 nm. (B) Chromatogram of a blank measurement (H2O) monitored at λ = 214 nm. Paenilamicin shows practically no retention at the C18 column.

Table S1. Homology analysis of the pam gene cluster products.

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