Casein peptone

To isolate DDPs produced by the transformants, we picked up the transformants that produced target DDPs the most and performed scaling up cultivation of the transformant. Each A. oryzae transformant was cultivated in 1/2 CPS medium (1.75% Czapek-Dox broth, 0.15% casein peptone (Nacalai Tesque), 0.15% meat peptone (Nacalai Tesque), 0.15% soy peptone, 1% soluble starch (Nacalai Tesque), 0.5% maltose monohydrate (Nacalai Tesque), 0.01% adenine (for adenine auxotrophic strains) for 1 week at 30 °C, and the culture media and mycelium were separated by filtration. The culture media was extracted with EtOAc two times, and the organic layer were combined and concentrated in vacuo to give crude extract. The freeze-dried mycelium was extracted with EtOAc (20% MeOH), and the extract was concentrated in vacuo. The mycelia extract was then dissolved in MeOH (0.1% water), washed with hexane three times, concentrated in vacuo to give crude extract. Each of the crude extract of culture and mycelium was fractionated by silica gel column chromatography, and subjected to further purification by flash chromatography and preparative TLC (SiO₂ or ODS). All the DDP structures were fully determined by spectral analyses (Supplementary Figs. 8–10, 14, 15, 18, 19, 22, 23, 26, 27, 32, 33, 37, 38, 41, 42, 52, 54–77, Supplementary Tables 7–18, 25 and Supplementary Notes 3–6, 8, 10, 12, 14, 18, 20, 22, 31).