TABLE 2.
Overview of engineering strategies and their indicated influence on small peptide production.
| Target peptide | Signal sequence | Strain engineering | Relative densitometric quantification | Absolute quantification via MS |
|---|---|---|---|---|
| 3×FLAG | Ost1pre | WT | ∼100% | |
| | Ost1pre | P HHX1 -SEC61 | ∼200% | |
| | Ost1pre | P HHX1 -SEC62 | ∼100% | |
| | Ost1pre | P HHX1 -SEC63 | <100% | |
| | Ost1pre | P HHX1 -KAR2 | <100% | |
| | Ost1pre | SEC62-P HHX1 -SEC61 | ∼200% | |
| | MFαpre-Ost1pre | WT | ∼800% | |
| | Ost1pre-Scw10pro | WT | ∼1600% | |
| | Ost1pre | aqr1Δ | ∼600% | |
| | Ost1pre | fms1-2Δ | ∼600% | |
| | Ost1pre | skg3Δ | ∼500% | |
| | Ost1pre | tCDC48 | ∼900% | 0.8 mg/L |
| dQ-brazzein | Ost1pre | WT | ∼100% | |
| | Ost1pre | aqr1Δ | ∼200% | |
| | Ost1pre | fms1-2Δ | ∼200% | 140 mg/L |
| | Ost1pre | skg3Δ | ∼200% | |
| | Ost1pre | tCDC48 | ∼100% | |
| Linker-sCT | Ost1pre-Scw10pro | WT | ∼100% | |
| | Ost1pre-Scw10pro | aqr1Δ | ∼200% | |
| | Ost1pre-Scw10pro | fms1-2Δ | ∼200% | |
| | Ost1pre-Scw10pro | tCDC48 | ∼300% | |
| Linker-α-MSH | Ost1pre-Scw10pro | WT | ∼100% | |
| | Ost1pre-Scw10pro | aqr1Δ | ∼200% | |
| | Ost1pre-Scw10pro | fms1-2Δ | ∼200% | 0.2 μg/L |
| | Ost1pre-Scw10pro | tCDC48 | ∼100% | |
Relative quantification is shown relative to the respective parental production strain, together with absolute quantification by HPLC–MS for the best-performing strain of each peptide. Relative quantification of dQ-brazzein was based on Coomassie-stained SDS-PAGE and densitometric analysis of two biological replicates ( ). The remaining peptides were analyzed by immunoblotting using two biological replicates ( ).