Fig. 3
Adaptive laboratory evolution to increase glycerol conversion. (A) Inoculation scheme of a manual ALE in shake flasks. OD-values for a repetitive batch experiment (11 cycles) performed in shake flasks. U. maydis MB215 ∆cyp3 ∆MEL ∆UA ∆dgat ∆Pria1::Petef ∆fuz7 PetefmttA was grown in defined MTM with 100 g L-1 pure glycerol as the sole carbon source and 2.4 g L-1 NH4Cl. (B) Verification experiment of the evolved strain compared to the original U. maydis strain in defined MTM containing 125 g L-1 pure glycerol and 2.4 g L-1 NH4Cl. The cultivation was performed in a BioLector Pro under the following conditions: 1400 rpm, 30 °C, 85% humidity, online measurements were taken every 10 min, sacrifice samples were taken in irregular intervals during the cultivation. Metabolites were analysed in duplicate via HPLC