Figure 5.

Modular coculturing of S. cerevisiae strains expressing BUP1 facilitates opiate production from l-DOPA. A, Each strain, transformed with a plasmid for BUP1 expression, was genomically integrated with the BIA biosynthetic genes indicated. Strains 1–3 were engineered to produce (S)-reticuline, thebaine, and codeine, respectively. “Binary” (strains 1 + 2) or “tertiary” (1 + 2 + 3) cocultures were fed l-DOPA and incubated for 24 h, after which media were profiled for (S)-reticuline, salutaridine, thebaine, and codeine. B and C, The same data but with differently scaled y axes to highlight quantitative data variations. 6OMT, norcoclaurine 6-O-methyltransferase; CNMT, coclaurine N-methyltransferase; 4′OMT, 3′-hydroxy-N-methylcoclaurine 4′-O-methyltransferase; CPR, cytochrome P450 reductase; COR, codeinone reductase. The se was calculated using four biological replicates representing four individual transformants. The number above each bar indicates the P value obtained using an unpaired, two-tailed Student’s t test to compare with appropriate empty vector controls.