Fig. 2

Comparison of E. coli BL21 (DE3) clones with single amino acid exchanges in a recombinant protein cultivated on synthetic Wilms-MOPS auto-induction medium containing 0.5 g L⁻¹ glucose, 2 g L⁻¹ lactose, 5 g L⁻¹ glycerol and 0.2 M MOPS buffer. All cultures were performed in parallel in one MTP. Each group of cultures consists of five replicates. According to their respiration behavior the clones were divided in Type A (wild type enzyme, D91R) and Type B (G93Y). a–c Online monitoring of the oxygen transfer rate (OTR, black lines) and the dissolved oxygen tension (DOT, blue lines). The phases (I–IV) typical for this kind of culture are indicated by dashed lines: (I) increase in OTR due to growth on glucose, at the end of this phase glucose is exhausted; (II) product (FbFP) formation during parallel consumption of lactose and glycerol; (III) increase in OTR due to growth on residual glycerol; (IV) end of culture. d–f Microbial growth monitored via scattered light at 650 nm (green lines) and the fluorescence intensity of FbFP (λ_ex = 450 nm; λ_em = 492 nm). Cultivation conditions: 0.1 g L⁻¹ infrared oxygen-sensitive nanoparticles; 48 round deep-well MTP without optodes, V_L = 800 µL, n = 1000 rpm, shaking diameter d₀ = 3 mm, 37 °C