3.2. Synthesis of 1-octanol
Given its fuel characteristics, we therefore set out to engineer a pathway for the production of 1-octanol. The previously engineered pathway, which resulted in the synthesis of longer chain fatty alcohols (C12-C16), consisted of E. coli acyl-ACP thioesterase TesA, M. marinum carboxylic acid reductase (CAR), B. subtilisphosphopantetheinyl transferase, (Sfp) and E. coli Ahr ( Akhtar et al., 2013). Since the chain length of the fatty alcohol was determined by the substrate specificity of TesA, this enzyme was therefore replaced with one that had previously been shown to generate the 8-carbon fatty acid precursor, octanoic acid ( Jing et al., 2011) ( Fig. 1a). Induction of the pathway in the presence of IPTG for 14 hours led to the synthesis of 29(±4) 1-hexanol mg L−1 and 62(±3) 1-octanol mg L−1 with an average yield of 12(±0.4) mg of 1-octanol g−1 glucose and a productivity of 4.4 mg (±0.04) mg 1-octanol L−1 h−1 ( Fig. 1b; Table 2). No other new metabolites were observed by gas chromatography–mass spectrometry (GC–MS) analysis.
3.2. Synthesis of 1-octanolGiven its fuel characteristics, we therefore set out to engineer a pathway for the production of 1-octanol. The previously engineered pathway, which resulted in the synthesis of longer chain fatty alcohols (C12-C16), consisted of E. coli acyl-ACP thioesterase TesA, M. marinum carboxylic acid reductase (CAR), B. subtilisphosphopantetheinyl transferase, (Sfp) and E. coli Ahr ( Akhtar et al., 2013). Since the chain length of the fatty alcohol was determined by the substrate specificity of TesA, this enzyme was therefore replaced with one that had previously been shown to generate the 8-carbon fatty acid precursor, octanoic acid ( Jing et al., 2011) ( Fig. 1a). Induction of the pathway in the presence of IPTG for 14 hours led to the synthesis of 29(±4) 1-hexanol mg L−1 and 62(±3) 1-octanol mg L−1 with an average yield of 12(±0.4) mg of 1-octanol g−1 glucose and a productivity of 4.4 mg (±0.04) mg 1-octanol L−1 h−1 ( Fig. 1b; Table 2). No other new metabolites were observed by gas chromatography–mass spectrometry (GC–MS) analysis.
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