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Fig. 7 | Journal of Biological Engineering

Fig. 7

From: Cellulolytic RoboLector – towards an automated high-throughput screening platform for recombinant cellulase expression

Fig. 7

Auto-induction medium variation for E. coli expressing recombinant extremophile endoglucanase cel8H from Halomonas sp. tagged with fluorescent mCherry applying the RoboLector system. Cultivation and on-line monitoring of microbial growth (via scattered light, a) and recombinant cellulase expression (via mCherry fluorescence, b) for different auto-induction medium compositions (see Table 2). c Final mCherry fluorescence and volumetric cellulase activity applying the PAHBAH reducing sugar assay for different auto-induction medium compositions. Error bars indicate standard deviation of three simultaneously performed assays. Cultivation conditions: 96well MTP, TB medium with glycerol (5 g L−1) and varied glucose (0.5–3 g L−1) and lactose (2–10 g L−1) concentrations, VL = 200 μl, n = 1000 rpm, d0 = 3 mm, 37 °C. Hydrolysis conditions: incubation in 96well DWP for 60 min, VL = 200 μL, 10 g L−1 CMC in 0.1 M acetate buffer, pH = 5, 45 °C, n = 900 rpm, d0 = 3 mm. PAHABH assay conditions: incubation in 96well DWP for 10 min, VL = 225 μL, n = 900 rpm, d0 = 3 mm, 85 °C, absorbance measurement at 410 nm

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