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. Author manuscript; available in PMC: 2021 Jun 19.
Published in final edited form as: ACS Synth Biol. 2020 May 7;9(6):1349–1360. doi: 10.1021/acssynbio.0c00038

Figure 3.

Figure 3.

In vitro TT-assay and production of hapalindoles. Top: Biosynthetic gene cluster of Fischerella ambigua UTEX 1903 (fam). The genes for transporters (white, A1–3), oxygenases (blue, B1–6), cyclases (pink, C1–5), aromatic prenyltransferases (red, D1–2), GPP synthases (cyan, E1–5), regulators (yellow, F1–2), cis-indole isonitrile synthase (green, H1–3) and other core enzymes (black, G1–2) are shown. Bottom: Extracted ion chromatograms (EICs) are shown in all panels. Detection of in vitro synthesized hapalindole products showing EIC (m/z of 305) with no plasmid (A), FamD2 alone (B) for cis-indole isonitrile 3, FamD2 plus FamC1 (C) for 12-epi-hapalindole U 4, FamD2 plus FamC2 and FamC3 (E) for hapalindole H 6, and FamD2 plus FamC1 and FamC4 (G) for hapalindole U 5, and 12-epi-hapalindole U 4 production. The traces D, F, and H are the authentic standards of 12-epi-hapalindole U 4, hapalindole H 6, and hapalindole U 5, respectively.