Table 3. Total number of transformants obtained with vectors containing pleuromutilin pathway genes under the control of A. bisporus gpdII promoter.
| Gene Expressed | Plasmid | Total transformants | Pleuromutilin titre range (μg/g) | Clearing zone range (and average) as a percentage of wild-type |
|---|---|---|---|---|
| FDS | p004-CP1-FDSgene | 55 | 595–810 | – |
| FDS | p004-CP1-iFDSgene | 20 | 600–920 | – |
| GGS | p004-CP1-GGSgene | 32 | 495–1,010 | – |
| GGS | p004-CP1-iGGSgene | 22 | 605–795 | – |
| Cyclase | pYES-hph-Cycgene | 31 | – | 0–109.4% (89.0%) |
| Cyclase | pYES-hph-iCycgene | 23 | – | 100.8–122.0% (106.7%) |
| P450-1 | pYES-hph-p450-1gene | 20 | – | 70.4–103.0% (92.4%) |
| P450-1 | pYES-hph-ip450-1gene | 23 | – | 77.0–103.7% (92.5%) |
| P450-2 | pYES-hph- p450-2gene | 22 | – | 68.1–107.3% (89.3%) |
| P450-2 | pYES-hph-ip450-2gene | 30 | – | 83.7–105.2% (94.2%) |
| P450-3 | pYES-hph-ip450-3gene | 32 | – | 49.7–113.8% (98.3%) |
The FDS and GGS lines were analysed by HPLC. All others were analysed by bioassay. The titres of pleuromutilin recorded through HPLC (μg of pleuromutilin per g of dry mycelial mass) are reported for the transformant lines overexpressing the genes fds and ggs. The wild-type levels were 690–815 μg/g.