Fig. 3 | Scientific Reports

Fig. 3

From: A robust high-throughput screening system to assess bacterial tyrosine ammonia lyase activity in the context of tyrosine inherited metabolic disorders

Fig. 3

(a) Analysis of the supernatant fraction of the bacterial cell lysate, containing FjTAL, by SDS-PAGE. E. coli BL21 (DE3) pET42b( +)-FjTAL cultures were incubated at four different temperatures (16 °C, 22 °C, 30 °C and 37 °C) post-induction of FjTAL expression followed by sampling after 0 h, 2 h, 4 h, 6 h, 24 h and 48 h. FjTAL expression could be detected 2 h post IPTG addition at 16 °C, 22 °C and 30 °C. Almost no expression was observed after incubation at 37 °C. While the expression of FjTAL was stable at 37 °C, increased levels of protein expression were observed at the three other temperatures. Uncropped gels are available as Supplementary Information in Supplementary Fig. 1. (b) Analysis of the detected FjTAL bands on the SDS-PAGE gels. Normalized pixel intensity of the FjTAL band was calculated for each condition. After comparison, incubation for 24 h at 22 °C post induction of protein expression were chosen as ideal expression conditions.

Back to article page