Fig. 2
From: Metabolic engineering of Escherichia coli for the production of cinnamaldehyde
The construction of expression system, production of each enzymes and cinnamaldehyde in E. coli. a The schematic diagram of plasmid pHB-CAD for the expression of three synthesis genes (SmPAL, ScCCL, and AtCCR genes) under the IPTG-inducible trc promoter (Ptrc). RBS means the ribosome binding site for translation and restriction enzyme sites were designated. b Western blot analysis of gene expression under two different temperature (30 and 37 °C). For the detection of SmPAL (lanes 1–4), anti-FLAG-HRP antibody was used and for the detection of ScCCL and AtCCR (lanes 5–8), anti-His-HRP antibody was used. Lanes 1, 3, 5, and 7 indicate total protein fraction, and lanes 2, 4, 6, and 8 indicates the soluble protein fractions. Lanes 1, 2, 5, and 6 indicate samples at 30 °C, and lanes 3, 4, 7, and 8 indicate samples at 37 °C. Symbols: Closed arrowhead, SmPAL; open arrowhead, ScCCL; solid arrow, AtCCR. c HPLC analysis of cinnamaldehyde produced under two different temperatures. Error bars represent standard deviation of the mean (n = 3)