Fig. 6. In vivo biotransformations implicate NozPT as an indole alkaloid DKP prenylation catalyst.
a Heterologous expression of nozPT by S. lividans TK24 and complementation with DD-cWW (5) resulted in the production of prenylated 6, with [M + H]+ m/z 441 by ESI+. LC was conducted with a reversed-phase C18 column and gradient mobile phase of H2O/MeCN (40–90%) containing 0.1% HOAc, detailed in “Methods”. b Atomic connectivity of 6 was established by key COSY (blue) and 1H-13C HMBC (red) correlations. c Key ROESY correlations (in red) established the configuration of 6, with H-2’ and prenyl groups exo to H-9/H-9’; correlations in gray with X were absent or very weak compared to correlations in red. d The predicted mechanism for NozPT-mediated reactions includes DMAPP transfer onto 5 with C-N annulation to yield 6. Source data are provided as a Source Data file.