Fig. 1
From: Discovery of novel amino acid production traits by evolution of synthetic co-cultures
ALE of a CoNoS comprising two amino-acid auxotrophic strains. A ALE of the CoNoS composed of C. glutamicum ΔARG LEU++  ↔ ΔLEU ARG+. To start the ALE, both strains were cultivated in shake flask monocultures in CGXII medium with 2% (w/v) glucose and 3 mM of the required amino acid. After 2 days at 30 °C, the cells were washed and used in a 1:1 ratio based on OD600 measurements to inoculate the first batch. 16 repetitive batches were cultivated in CGXII medium with 2% (w/v) glucose and 100 µM IPTG in microtiter plates (MTPs) at 30 °C, 1400 rpm. Displayed is the on-line backscatter signal of batches 2–16. A backscatter threshold (dashed grey line) triggered cell suspension transfer to inoculate the following batch. One representative of three independent replicates is shown. B Specific growth rate calculation for the ALE shown in (A). C, D Each evolved strain (evo1, evo2, and evo3) was isolated from one independent replicate of the ALE and cultivated in biological triplicates with the corresponding non-evolved partner. WT monoculture and the CoNoS comprising the non-evolved strains are shown as reference cultivations. The CoNoS were cultivated in CGXII medium with 2% (w/v) glucose and incubated in MTPs at 30 °C, 1400 rpm. Mean values and standard deviations are shown as lines and shaded areas, respectively